Stereological study and analysis of oxidative stress during renal aging in rats

Purpose To evaluate renal histological changes by stereology and morphometry and analyze the main markers of oxidative stress in rats undergoing natural aging. Methods Seventy two Wistar rats were divided into six groups of 12 rats each, which were euthanized at 3, 6, 9, 12, 18, and 24 months of age. Right kidney was stereologically and morphometrically analyzed to calculate the volumetric density (Vv[glom]), numerical density (Nv[glom]) and glomerular volume (Vol[glom]). Left kidney was used to determine the levels of nonprotein thiols, lipid peroxidation, and protein carbonylation, as well as the activities of superoxide-dismutase and catalase enzymes. Results Both Vv[glom] and Nv[glom] values showed gradual decreases between groups. Activity of superoxide-dismutase was elevated at 24 months of age, and the levels of nonprotein thiols were higher in older animals. Greater catalase activity and protein carbonylation were observed in animals between 6 and 12 months of age but lessened in older rats. Lipid peroxidation decreased in the older groups. Conclusions Morphometric and stereological analyses revealed a gradual decrease in the volume and density of renal glomeruli during aging, as well as kidney atrophy. These findings related to oxidative stress clarify many changes occurring in kidney tissues during senescence in rats.

Stereological study and analysis of oxidative stress during renal aging in rats

Purpose To evaluate renal histological changes by stereology and morphometry and analyze the main markers of oxidative stress in rats undergoing natural aging. Methods Seventy two Wistar rats were divided into six groups of 12 rats each, which were euthanized at 3, 6, 9, 12, 18, and 24 months of age. Right kidney was stereologically and morphometrically analyzed to calculate the volumetric density (Vv[glom]), numerical density (Nv[glom]) and glomerular volume (Vol[glom]). Left kidney was used to determine the levels of nonprotein thiols, lipid peroxidation, and protein carbonylation, as well as the activities of superoxide-dismutase and catalase enzymes. Results Both Vv[glom] and Nv[glom] values showed gradual decreases between groups. Activity of superoxide-dismutase was elevated at 24 months of age, and the levels of nonprotein thiols were higher in older animals. Greater catalase activity and protein carbonylation were observed in animals between 6 and 12 months of age but lessened in older rats. Lipid peroxidation decreased in the older groups. Conclusions Morphometric and stereological analyses revealed a gradual decrease in the volume and density of renal glomeruli during aging, as well as kidney atrophy. These findings related to oxidative stress clarify many changes occurring in kidney tissues during senescence in rats.(AU)

Kidney anatomy, histology and histometric traits associated to renosomatic index in Gymnotus inaequilabiatus (Gymnotiformes: Gymnotidae)

The Gymnotus inaequilabiatus is a Neotropical fish widely distributed in marginal areas of bays. The aim of this study was to describe the main histological and histopathological traits in the head and exocrine kidney. Here, histometric and structural density techniques were associated with renosomatic index (RSI). The kidney was processed for light microscopy. Lipofuscin and hemosiderin content were visually estimated in the melonomacrophages centers (MMCs). All the biometric body variables were correlated with RSI, especially the kidney weight and gross lesions count. The general architecture of head and exocrine kidney was similar to that described for other teleost species. MMCs were prevalent in both portions and correlated with RSI in the head and exocrine kidney. Granulomatous structures were often observed in both portions; however, they were associated only in the exocrine kidney with RSI. Of all the structures hystometrically estimated, only proximal tubular diameter and thickness, and distal tubular thickness were correlated to renosomatic index. The RSI is an useful biometric variable that represent some physiological and morphological characteristics of kidney in G. inaequilabiatus. These findings may be used in future studies to evaluate the effects of environmental stressors on the renal adaptative physiological process.(AU)

Gymnotus inaequilabiatus é um peixe neotropical amplamente distribuído em áreas marginais de baías. O objetivo deste estudo foi descrever características anatômicas, histológicas e histopatológicas no rim cefálico e exócrino. Análises histométricas e a densidade volumétrica estrutural foram associadas ao índice renossomático (IRS). Fragmentos de ambos tecidos foram processados para análise e microscopia de campo claro. O conteúdo de lipofuscina e hemossiderina foi estimado visualmente nos centros de melanomacrófagos (CMMs). Todas as variáveis corporais biométricas foram correlacionadas com o IRS, principalmente o peso renal e a contagem de lesões superficiais. A arquitetura geral do rim cefálico e exócrino foi semelhante à descrita para outras espécies de teleósteos. Os CMMs foram frequentes em ambas as porções e correlacionaram-se com o IRS. Estruturas granulomatosas foram frequentemente observadas em ambas porções; no entanto, foram associadas ao IRS apenas no rim exócrino. De todas as estruturas estimadas histometricamente, apenas o diâmetro e espessura dos túbulos proximais e distais correlacionaram-se ao IRS. Este índice mostrou-se uma variável útil associada a características fisiológicas e morfológicas do rim de G. inaequilabiatus. Esses achados podem ser empregados em futuros estudos a fim de avaliar efeitos ambientais bem como processos adaptativos fisiológicos.(AU)

Kidney anatomy, histology and histometric traits associated to renosomatic index in Gymnotus inaequilabiatus (Gymnotiformes: Gymnotidae)

The Gymnotus inaequilabiatus is a Neotropical fish widely distributed in marginal areas of bays. The aim of this study was to describe the main histological and histopathological traits in the head and exocrine kidney. Here, histometric and structural density techniques were associated with renosomatic index (RSI). The kidney was processed for light microscopy. Lipofuscin and hemosiderin content were visually estimated in the melonomacrophages centers (MMCs). All the biometric body variables were correlated with RSI, especially the kidney weight and gross lesions count. The general architecture of head and exocrine kidney was similar to that described for other teleost species. MMCs were prevalent in both portions and correlated with RSI in the head and exocrine kidney. Granulomatous structures were often observed in both portions; however, they were associated only in the exocrine kidney with RSI. Of all the structures hystometrically estimated, only proximal tubular diameter and thickness, and distal tubular thickness were correlated to renosomatic index. The RSI is an useful biometric variable that represent some physiological and morphological characteristics of kidney in G. inaequilabiatus. These findings may be used in future studies to evaluate the effects of environmental stressors on the renal adaptative physiological process.(AU)

Gymnotus inaequilabiatus é um peixe neotropical amplamente distribuído em áreas marginais de baías. O objetivo deste estudo foi descrever características anatômicas, histológicas e histopatológicas no rim cefálico e exócrino. Análises histométricas e a densidade volumétrica estrutural foram associadas ao índice renossomático (IRS). Fragmentos de ambos tecidos foram processados para análise e microscopia de campo claro. O conteúdo de lipofuscina e hemossiderina foi estimado visualmente nos centros de melanomacrófagos (CMMs). Todas as variáveis corporais biométricas foram correlacionadas com o IRS, principalmente o peso renal e a contagem de lesões superficiais. A arquitetura geral do rim cefálico e exócrino foi semelhante à descrita para outras espécies de teleósteos. Os CMMs foram frequentes em ambas as porções e correlacionaram-se com o IRS. Estruturas granulomatosas foram frequentemente observadas em ambas porções; no entanto, foram associadas ao IRS apenas no rim exócrino. De todas as estruturas estimadas histometricamente, apenas o diâmetro e espessura dos túbulos proximais e distais correlacionaram-se ao IRS. Este índice mostrou-se uma variável útil associada a características fisiológicas e morfológicas do rim de G. inaequilabiatus. Esses achados podem ser empregados em futuros estudos a fim de avaliar efeitos ambientais bem como processos adaptativos fisiológicos.(AU)

Hepatoprotective and antineoplastic potencial of red propolis produced by the bees Apis mellifera in the semiarid of Rio Grande do Norte, Brazil

ABSTRACT: The objective of this study was to evaluate the hepatoprotective effect of the honey bee Apis mellifera ethanolic extract of the red propolis, obtained in four municipalities of the Rio Grande do Norte semi-arid region, through an in vitro evaluation of the antineoplastic potential in human hepatic carcinoma (HepG2) and normal cell lines (L929), and from the comet assay in hepatic cell lines (ZF-L hepatocytes) to evaluate the genoprotective potential of the extract. The hepatoprotective effect was also evaluated in vivo by the induction of chronic experimental hepatic lesions in rodents (Rattus norvegicus Berkenhout, 1769), Wistar line, by intraperitoneal administration of thioacetamide (TAA) at the dose of 0.2g/kg. The animals were distributed in the following experimental groups: G1 (control), G2 (treated with 500mg/kg ethanolic extract of propolis), G3 (treated with 500mg/kg of ethanolic extract and TAA) and G4 (treated with TAA). All rats were submitted to serum biochemical, macroscopic, histological and stereological biochemical exams of the liver. It was verified the genoprotective effect of red propolis since the mean damages promoted to DNA in cells tested with the extract were significantly lower than the mean of the positive control damage (hydrogen peroxide). The red propolis extract did not present cytotoxic activity to the tumor cells of human liver cancer, as well as to normal ones. The absence of cytotoxicity in normal cells may indicate safety in the use of the propolis extract. The results of the serum biochemical evaluation showed that the serum levels of the aminotransferase enzymes (AST) did not differ significantly between G1, G2 and G3 when compared to each other. G4 showed significant increase in levels compared to the other groups, indicating that the administration of the extract did not cause liver toxicity, as well as exerted hepatoprotective effect against the hepatic damage induced by TAA. The G3 and G4 animals developed cirrhosis, but in G3 the livers were characterized by the presence of small regenerative nodules and level with the surface of the organ, whereas in G4 the livers showed large regenerative nodules. The livers of the G1 and G2 animals presented normal histological appearance, whereas the livers of the G3 animals showed regenerative nodules surrounded by thin septa of connective tissue, and in G4 the regenerative nodules were surrounded by thick septa fibrous connective tissue. The analysis of the hepatic tissues by means of stereology showed that there was no statistical difference between the percentage of hepatocytes, sinusoids, and collagens in G1 and G2. In G3 the percentage of hepatocytes, sinusoids, and collagen did not differ significantly from the other groups. It was concluded that the ethanolic extract of the red propolis exerted a hepatoprotective effect, because it promoted in vitro reduction of the damage to the DNA of liver cells, antineoplastic activity in human hepatocellular carcinoma cell line (HepG2) and did not exert cytotoxic effect in normal cells or was able to reduce liver enzyme activity and the severity of cirrhosis induced by TAA in vivo.

RESUMO: Este estudo objetivou avaliar o efeito hepatoprotetor do extrato etanólico da própolis vermelha da abelha Apis mellifera, obtido em quatro municípios do semiárido do Rio Grande do Norte, mediante avaliação in vitro do potencial antineoplásico em linhagens de células de carcinoma hepático humano (HepG2) e em linhagens de células normais (L929), além do ensaio cometa em linhagens de células hepáticas (hepatócitos ZF-L) para avaliar o potencial genoprotetor do extrato. O efeito hepatoprotetor também foi avaliado in vivo através da indução de lesões hepática experimental crônica em roedores da espécie Rattus norvegicus (Berkenhout, 1769), linhagem Wistar, pela administração intraperitoneal de tioacetamida (TAA) na dose de 0,2g/kg. Os animais foram distribuídos nos seguintes grupos experimentais: G1 (controle), G2 (tratados com 500mg/kg de extrato etanólico da própolis), G3 (tratados com 500mg/kg de extrato etanólico e TAA) e G4 (tratados com TAA). Todos os ratos foram submetidos aos exames bioquímico sérico, anatomopatológico macroscópico, histológico e esteriológico do fígado. Foi constatado o efeito genoprotetor da própolis vermelha uma vez que as médias dos danos promovidos ao DNA em células testadas com o extrato foram significativamente inferiores à média dos danos do controle positivo (peróxido de hidrogênio). O extrato da própolis vermelha não apresentou atividade citotóxica para células tumorais de câncer de fígado humano, bem como para normais. A ausência de citotoxicidade em células normais, tal como constatado, pode indicar segurança no uso do extrato da própolis. Os resultados da avaliação bioquímica sérica demonstraram que os níveis séricos das enzimas aminotransferase (AST) não diferiram significativamente entre G1, G2 e G3, quando comparadas entre si. No G4 houve aumento significativo dos níveis em relação aos demais grupos, indicando que a administração do extrato não causou toxicidade hepática, bem como exerceu efeito hepatoprotetor frente ao dano hepático induzido pela TAA. Os animais dos G3 e G4 desenvolveram cirrose, porém no G3 os fígados caracterizaram-se pela presença de pequenos nódulos regenerativos e nivelados com a superfície do órgão, enquanto que no G4 os fígados apresentaram grandes nódulos regenerativos. Os fígados dos animais G1 e G2 apresentaram aspecto histológico normal, enquanto que os fígados dos animais do G3 apresentaram nódulos regenerativos circundados por finos septos de tecido conjuntivo, e nos do G4 os nódulos regenerativos foram circundados por espessos septos de tecido conjuntivo fibroso. A análise dos tecidos hepáticos por meio de estereologia mostrou que não houve diferença estatística entre o percentual de hepatócitos, sinusoides e colágenos nos G1 e G2. No G3 o percentual de hepatócitos, sinusoides e colágeno não diferiu significativamente dos demais grupos. Concluiu-se que o extrato etanólico da própolis vermelha exerceu efeito genoprotetor, por promover in vitro redução do dano ao DNA de células hepáticas, atividade antineoplásica em linhagem celular de carcinoma hepatocelular humano (HepG2) e não exerceu efeito citotóxico em células normais ou efeito hepatoprotetor in vivo com diminuição da gravidade da cirrose induzida por TAA.

Hepatoprotective and antineoplastic potencial of red propolis produced by the bees Apis mellifera in the semiarid of Rio Grande do Norte, Brazil / Potencial hepatoprotetor e antineoplásico da própolis vermelha produzida pela abelha Apis mellifera no semiárido do Rio Grande do Norte, Brasil

The objective of this study was to evaluate the hepatoprotective effect of the honey bee Apis mellifera ethanolic extract of the red propolis, obtained in four municipalities of the Rio Grande do Norte semi-arid region, through an in vitro evaluation of the antineoplastic potential in human hepatic carcinoma (HepG2) and normal cell lines (L929), and from the comet assay in hepatic cell lines (ZF-L hepatocytes) to evaluate the genoprotective potential of the extract. The hepatoprotective effect was also evaluated in vivo by the induction of chronic experimental hepatic lesions in rodents (Rattus norvegicus Berkenhout, 1769), Wistar line, by intraperitoneal administration of thioacetamide (TAA) at the dose of 0.2g/kg. The animals were distributed in the following experimental groups: G1 (control), G2 (treated with 500mg/kg ethanolic extract of propolis), G3 (treated with 500mg/kg of ethanolic extract and TAA) and G4 (treated with TAA). All rats were submitted to serum biochemical, macroscopic, histological and stereological biochemical exams of the liver. It was verified the genoprotective effect of red propolis since the mean damages promoted to DNA in cells tested with the extract were significantly lower than the mean of the positive control damage (hydrogen peroxide). The red propolis extract did not present cytotoxic activity to the tumor cells of human liver cancer, as well as to normal ones. The absence of cytotoxicity in normal cells may indicate safety in the use of the propolis extract. The results of the serum biochemical evaluation showed that the serum levels of the aminotransferase enzymes (AST) did not differ significantly between G1, G2 and G3 when compared to each other. G4 showed significant increase in levels compared to the other groups, indicating that the administration of the extract did not cause liver toxicity, as well as exerted hepatoprotective effect against the hepatic damage induced by TAA. The G3 and G4 animals developed cirrhosis, but in G3 the livers were characterized by the presence of small regenerative nodules and level with the surface of the organ, whereas in G4 the livers showed large regenerative nodules. The livers of the G1 and G2 animals presented normal histological appearance, whereas the livers of the G3 animals showed regenerative nodules surrounded by thin septa of connective tissue, and in G4 the regenerative nodules were surrounded by thick septa fibrous connective tissue. The analysis of the hepatic tissues by means of stereology showed that there was no statistical difference between the percentage of hepatocytes, sinusoids, and collagens in G1 and G2. In G3 the percentage of hepatocytes, sinusoids, and collagen did not differ significantly from the other groups. It was concluded that the ethanolic extract of the red propolis exerted a hepatoprotective effect, because it promoted in vitro reduction of the damage to the DNA of liver cells, antineoplastic activity in human hepatocellular carcinoma cell line (HepG2) and did not exert cytotoxic effect in normal cells or was able to reduce liver enzyme activity and the severity of cirrhosis induced by TAA in vivo.(AU)

Este estudo objetivou avaliar o efeito hepatoprotetor do extrato etanólico da própolis vermelha da abelha Apis mellifera, obtido em quatro municípios do semiárido do Rio Grande do Norte, mediante avaliação in vitro do potencial antineoplásico em linhagens de células de carcinoma hepático humano (HepG2) e em linhagens de células normais (L929), além do ensaio cometa em linhagens de células hepáticas (hepatócitos ZF-L) para avaliar o potencial genoprotetor do extrato. O efeito hepatoprotetor também foi avaliado in vivo através da indução de lesões hepática experimental crônica em roedores da espécie Rattus norvegicus (Berkenhout, 1769), linhagem Wistar, pela administração intraperitoneal de tioacetamida (TAA) na dose de 0,2g/kg. Os animais foram distribuídos nos seguintes grupos experimentais: G1 (controle), G2 (tratados com 500mg/kg de extrato etanólico da própolis), G3 (tratados com 500mg/kg de extrato etanólico e TAA) e G4 (tratados com TAA). Todos os ratos foram submetidos aos exames bioquímico sérico, anatomopatológico macroscópico, histológico e esteriológico do fígado. Foi constatado o efeito genoprotetor da própolis vermelha uma vez que as médias dos danos promovidos ao DNA em células testadas com o extrato foram significativamente inferiores à média dos danos do controle positivo (peróxido de hidrogênio). O extrato da própolis vermelha não apresentou atividade citotóxica para células tumorais de câncer de fígado humano, bem como para normais. A ausência de citotoxicidade em células normais, tal como constatado, pode indicar segurança no uso do extrato da própolis. Os resultados da avaliação bioquímica sérica demonstraram que os níveis séricos das enzimas aminotransferase (AST) não diferiram significativamente entre G1, G2 e G3, quando comparadas entre si. No G4 houve aumento significativo dos níveis em relação aos demais grupos, indicando que a administração do extrato não causou toxicidade hepática, bem como exerceu efeito hepatoprotetor frente ao dano hepático induzido pela TAA. Os animais dos G3 e G4 desenvolveram cirrose, porém no G3 os fígados caracterizaram-se pela presença de pequenos nódulos regenerativos e nivelados com a superfície do órgão, enquanto que no G4 os fígados apresentaram grandes nódulos regenerativos. Os fígados dos animais G1 e G2 apresentaram aspecto histológico normal, enquanto que os fígados dos animais do G3 apresentaram nódulos regenerativos circundados por finos septos de tecido conjuntivo, e nos do G4 os nódulos regenerativos foram circundados por espessos septos de tecido conjuntivo fibroso. A análise dos tecidos hepáticos por meio de estereologia mostrou que não houve diferença estatística entre o percentual de hepatócitos, sinusoides e colágenos nos G1 e G2. No G3 o percentual de hepatócitos, sinusoides e colágeno não diferiu significativamente dos demais grupos. Concluiu-se que o extrato etanólico da própolis vermelha exerceu efeito genoprotetor, por promover in vitro redução do dano ao DNA de células hepáticas, atividade antineoplásica em linhagem celular de carcinoma hepatocelular humano (HepG2) e não exerceu efeito citotóxico em células normais ou efeito hepatoprotetor in vivo com diminuição da gravidade da cirrose induzida por TAA.(AU)

The effect of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) on mice myocardial morphology / O efeito do 1-metil-4-fenil-1, 2, 3, 6 tetrahidropiridina (MPTP) na morfologia do miocárdio de camundongo

Studies have demonstrated sympathetic cardiac denervation in the MPTP mouse model. MPTP toxicity causes sympathetic nerve damage and depletion of heart norepinephrine. Previous evaluations of impairments in heart innervation have been based on imaging, electrophysiological and biochemical methods. However, these studies lacked information that can be obtained from morphoquantitative analyses. Thus, this study aimed to apply a design-based stereological method for evaluating the morphoquantitative alterations of myocardium following treatment with the neurotoxin MPTP in the C57/BL mouse. Our results showed that MPTP reduced the number of cardiomyocytes in the left ventricle.(AU)

Estudos têm demonstrado a desnervação simpática cardíaca no modelo da administração do MPTP em camundongo. A toxicidade do MPTP causa lesão ao nervo simpático e depleção da norepinefrina. As avaliações dos danos na inervação do coração são baseadas em métodos de imagem, eletrofisiológico e bioquímico. Contudo, estes estudos carecem de informações provenientes de análises morfoquantitativas. Assim, objetivou-se aplicar métodos estereológicos para avaliar as alterações morfoquantitativas do miocárdio após o tratamento com a neurotoxina MPTP no camundongo C57/BL. Nossos resultados mostraram que o MPTP causa redução no número de cardiomiócitos no ventrículo esquerdo.(AU)

The effect of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) on mice myocardial morphology / O efeito do 1-metil-4-fenil-1, 2, 3, 6 tetrahidropiridina (MPTP) na morfologia do miocárdio de camundongo

Studies have demonstrated sympathetic cardiac denervation in the MPTP mouse model. MPTP toxicity causes sympathetic nerve damage and depletion of heart norepinephrine. Previous evaluations of impairments in heart innervation have been based on imaging, electrophysiological and biochemical methods. However, these studies lacked information that can be obtained from morphoquantitative analyses. Thus, this study aimed to apply a design-based stereological method for evaluating the morphoquantitative alterations of myocardium following treatment with the neurotoxin MPTP in the C57/BL mouse. Our results showed that MPTP reduced the number of cardiomyocytes in the left ventricle.(AU)

Estudos têm demonstrado a desnervação simpática cardíaca no modelo da administração do MPTP em camundongo. A toxicidade do MPTP causa lesão ao nervo simpático e depleção da norepinefrina. As avaliações dos danos na inervação do coração são baseadas em métodos de imagem, eletrofisiológico e bioquímico. Contudo, estes estudos carecem de informações provenientes de análises morfoquantitativas. Assim, objetivou-se aplicar métodos estereológicos para avaliar as alterações morfoquantitativas do miocárdio após o tratamento com a neurotoxina MPTP no camundongo C57/BL. Nossos resultados mostraram que o MPTP causa redução no número de cardiomiócitos no ventrículo esquerdo.(AU)

Stereological and morphometric study of type 3 collagen formation in the cutaneous wounds of diabetic mice treated with mesenchymal stem cells

Introduction: Wound healing is a progressive, essential and complex physiological process that occurs as a restorative response after a tissue injury. It involves three phases: inflammation, proliferation and maturation. Exogenous, endogenous and pathological factors may interfere in the cicatricial process in humans and animals by altering the balance between the synthesis, degradation and remodelling of collagen and elastic fibres. Diabetes mellitus is a progressive metabolic disease that alters elastogenesis and collagenesis and induces delays in the healing process. Scientific evidence suggests that mesenchymal stem cells modulate the cicatricial response. Thus the objective of this work was to perform stereological and morphometric analysis to determine the formation of dermal fibres in cutaneous fragments of a murine model of diabetes mellitus.Materials, Methods & Results: Histological sections were obtained from the cutaneous wounds of diabetic mice. The cutaneous wounds were previously treated with autogenous mesenchymal stem cells, physiological solution or polyurethane membrane. The histological sections were subsequently processed and stained for type 1 and 3 collagen fibres and elastic fibres using Picrosirius Red and Weigert staining, respectively. Histological sections stained with Picrosirius Red presented three types of birefringence under polarised light microscopy that corresponded to red colours for type 1 collagen and green and yellow colours for type 3 collagen. Weigert staining presented three colours for histological structures under white light microscopy that corresponded to black colours for elastic fibres, variations in colour from pink to purple for other structures and dermal attachments. The elastic fibres, represented by a black colour, presented in a heterogeneous form and were either identified as thin, punctiform or rectangular fibres or as elastic agglomerates.[....]

Stereological and morphometric study of type 3 collagen formation in the cutaneous wounds of diabetic mice treated with mesenchymal stem cells

Introduction: Wound healing is a progressive, essential and complex physiological process that occurs as a restorative response after a tissue injury. It involves three phases: inflammation, proliferation and maturation. Exogenous, endogenous and pathological factors may interfere in the cicatricial process in humans and animals by altering the balance between the synthesis, degradation and remodelling of collagen and elastic fibres. Diabetes mellitus is a progressive metabolic disease that alters elastogenesis and collagenesis and induces delays in the healing process. Scientific evidence suggests that mesenchymal stem cells modulate the cicatricial response. Thus the objective of this work was to perform stereological and morphometric analysis to determine the formation of dermal fibres in cutaneous fragments of a murine model of diabetes mellitus.Materials, Methods & Results: Histological sections were obtained from the cutaneous wounds of diabetic mice. The cutaneous wounds were previously treated with autogenous mesenchymal stem cells, physiological solution or polyurethane membrane. The histological sections were subsequently processed and stained for type 1 and 3 collagen fibres and elastic fibres using Picrosirius Red and Weigert staining, respectively. Histological sections stained with Picrosirius Red presented three types of birefringence under polarised light microscopy that corresponded to red colours for type 1 collagen and green and yellow colours for type 3 collagen. Weigert staining presented three colours for histological structures under white light microscopy that corresponded to black colours for elastic fibres, variations in colour from pink to purple for other structures and dermal attachments. The elastic fibres, represented by a black colour, presented in a heterogeneous form and were either identified as thin, punctiform or rectangular fibres or as elastic agglomerates.[....](AU)

Stereology of spix's yellow-toothed cavy brain (Galea spixii, WAGLER, 1831)

Spix's Yellow-toothed Cavy is a histrichomorphic rodent of the Caviidae family found in South American countries such as Brazil and Bolivia. It is a widely-used species as an experimental model in research in reproductive biology due to morphological and reproductive characteristics, such as the similarity in the placental development of Galea spixii and human species. However, there are no studies on the behavior of this species or on its brain morphology. Considering the lack of information in the literature about the brain and internal structures of Galea spixii, this study aimed to stereologic evaluate the brain as well as the volumetric proportions of the hippocampus and corpus callosum. Therefore, ten healthy animals were used from the Wild Animal Multiplication Center of the Universidade Federal Rural do Semiárido. The brains were measured in terms of external length, height and width, followed by fixation in paraformaldehyde solution 4% and coronary cuts with a thickness of 4mm and the rostral face of each cut was photographed, with the images being analyzed to determine the volumetric proportions of the required areas. There was no statistically significant difference between the means of volume, length, width and height when the right and left hemispheres were compared, and there was also no statistically significant difference between the volume of the corpus callosum and hippocampus in both hemispheres. The brain of Galea spixii is larger when compared to the rodent Rattus norvegicus. Volumetric differences may be responsible for distinct behavioral aspects between these species.

Stereology of spix's yellow-toothed cavy brain (Galea spixii, WAGLER, 1831)

Spix's Yellow-toothed Cavy is a histrichomorphic rodent of the Caviidae family found in South American countries such as Brazil and Bolivia. It is a widely-used species as an experimental model in research in reproductive biology due to morphological and reproductive characteristics, such as the similarity in the placental development of Galea spixii and human species. However, there are no studies on the behavior of this species or on its brain morphology. Considering the lack of information in the literature about the brain and internal structures of Galea spixii, this study aimed to stereologic evaluate the brain as well as the volumetric proportions of the hippocampus and corpus callosum. Therefore, ten healthy animals were used from the Wild Animal Multiplication Center of the Universidade Federal Rural do Semiárido. The brains were measured in terms of external length, height and width, followed by fixation in paraformaldehyde solution 4% and coronary cuts with a thickness of 4mm and the rostral face of each cut was photographed, with the images being analyzed to determine the volumetric proportions of the required areas. There was no statistically significant difference between the means of volume, length, width and height when the right and left hemispheres were compared, and there was also no statistically significant difference between the volume of the corpus callosum and hippocampus in both hemispheres. The brain of Galea spixii is larger when compared to the rodent Rattus norvegicus. Volumetric differences may be responsible for distinct behavioral aspects between these species.(AU)

CICLO PILOSO E VOLUME FOLICULAR DÉRMICO DO ROEDOR SILVESTRE, CUTIA (Dasyprocta prymnolopha), CRIADO EM CATIVEIRO, NA REGIÃO NORDESTE DO BRASIL.

Os pelos são estruturas anexas da pele da maioria dos mamíferos e se desenvolvem a partir de estruturas pilosas primitivas, os folículos pilosos. A duração de cada fase do ciclo de crescimento piloso pode ser influenciada por vários fatores como genética, idade, sexo, região corporal, hormônios, estado de saúde e fatores climáticos. A dinâmica de desenvolvimento dos pelos é um assunto complexo ainda não totalmente elucidado ou caracterizado em todas as espécies animais. Não foram encontrados dados na literatura sobre o volume folicular e ciclo piloso do roedor silvestre cutia (Dasyprocta prymnolopha), portanto objetivou-se com este estudo avaliar o ciclo piloso durante o período de um ano, nas estações seca e chuvosa e comparar volume folicular dérmico das regiões interescapular, abdominal média lateral e femoral de cutias machos e fêmeas, criadas em cativeiro no município de Teresina-PI. As práticas deste experimento foram aprovadas pela CEUA (589/19) e SISBIO (70275-2). Foram utilizadas seis cutias sendo três machos e três fêmeas com idade variando entre 3 e 10 anos. As amostras de pelos para o tricograma foram coletadas a cada 30 dias, no período de outubro de 2019 a setembro de 2020. Os pelos foram obtidos das três regiões de interesse alternando-se o antímero a cada mês. Foram extraídos no sentido de crescimento com auxílio de uma pinça hemostática coberta com borracha na região denteada, posteriormente 100 pelos de cada região foram colados um a um em lâmina de microscopia para identificação das fases do ciclo piloso por meio da análise morfológica dos bulbos pilosos em microscopia óptica. Biópsias cutâneas foram coletadas das três áreas citadas, processadas com um ano de antecedência para minimizar o estresse nos animais (CEUA: 407/17; SISBIO: 60484-1) e foram utilizadas para a determinação do volume folicular dérmico através de análise dos cortes histológicos pelo método de estereologia. O número de interações entre malha e folículos pilosos foi gravado e contado através do software Image J. Os fatores climáticos (temperatura, radiação, fotoperíodo, insolação, umidade) da cidade de Teresina- PI foram obtidos do boletim metereológico da EMBRAPA. Realizou-se análise variância (ANOVA), seguido do teste de Tukey a 5% de probabilidade. Sob condições de cativeiro, em faixa climática tropical e região semiárida, cutias (Dasyprocta prymnolopha) saudáveis, machos e fêmeas, apresentam maior percentual de pelos em fase telogênica, durante todo o ano, independente do período seco (machos 98,8% e fêmeas 98,9 %) ou chuvoso (machos 96,8 % e fêmeas 98,3%). Machos e fêmeas não exibiram diferenças entre densidade pilosa, duração da fase do ciclo do pelo e volume folicular dérmico.

Hairs are structures attached to the skin of most mammals and develop from primitive hair structures, the hair follicles. The length of each phase of the hair growth cycle can be influenced by several factors such as genetics, age, sex, body region, hormones, health status and climatic factors. The dynamics of hair development is a complex issue that has not yet been fully elucidated or adapted in all animal species. No data were found in the literature on the follicular volume and hair cycle of the wild rodent agouti (Dasyprocta prymnolopha), so the aim of this study was to evaluate the hair cycle over a period of one year, in the dry and rainy seasons and to compare dermal follicular volume of the interscapular, lateral abdominal and femoral regions of the male agouti and elevated, elevated in captivity in the municipality of Teresina-PI. As the experimental practices were approved by CEUA (589/19) and SISBIO (70275-2). Six agouti, three males and three defined, aged between 3 and 10 years, were used. The nostalgia of the hairs for the trichogram were collected every 30 days, from October 2019 to September 2020. The hairs were chosen from the three regions of interest, alternating the antimere every month. They were extracted in the direction of growth with the aid of hemostatic forceps covered with rubber in the dentate region, then 100 hairs from each region were glued one by one on a microscope slide to identify the phases of the hair cycle through the morphological analysis of the hair bulbs in optical microscopy. Skin biopsies were collected from the three areas mentioned, processed one year in advance to minimize stress on the animals (CEUA: 407/17; SISBIO: 60484-1) and were used to determine the dermal follicular volume, through analysis of the sections histological methods by the stereology method. The number of interactions between mesh and hair follicles was recorded and counted using Image J software. Climatic factors (temperature, radiation, photoperiod, insolation, humidity) in the city of Teresina- PI were obtained from the EMBRAPA weather report. Analysis of variance (ANOVA) was performed, followed by Tukey's test at 5% probability. Under conditions of captivity, in tropical climate range and semiarid region, healthy agouti (Dasyprocta prymnolopha) healthy, male and female, have a higher percentage of telogenic hair throughout the year, regardless of the dry period (98.8% males and females 98.9%) or rainy (male 96.8% and female 98.3%). Males and females did not show differences between hair density, length of the hair cycle and dermal follicular volume.

Alterações da microestrutura dos gânglios cervical cranial e trigeminal decorrentes de modelo experimental da Síndrome de Horner

A síndrome de Horner (SH) é uma afecção decorrente da lesão ou ruptura da inervação simpática oftálmica. Ocorre em várias espécies animais, inclusive a humana e os sinais clínicos mais comuns são enoftalmia, anisocoria, ptose palpebral e protrusão da terceira pálpebra ipsilateral à lesão. O presente estudo objetivou avaliar possíveis alterações morfoquantitativas decorrentes da SH induzida em ratos Wistar. Foram utilizados 12 ratos machos divididos em três grupos de quatro indivíduos, sendo eles: grupo I (G1): controle, sem indução da SH, grupo II (G2): eutanásia após duas semanas da indução da SH e grupo III (G3): eutanásia após sete semanas da indução. A indução foi realizada cirurgicamente, escolhendo de forma aleatória o antímero da lesão. Para tal, os animais passaram por protocolos anestésicos e analgésicos, e após as cirurgias, semanalmente, por avaliação clínica oftálmica para acompanhamento dos sinais clínicos. Após os tempos determinados de eutanásia, foi realizada coleta do material biológico para posterior avaliação quantitativa (avaliações estereológicas referentes ao volume total, densidade de volume e volume total de neurônios) e qualitativa dos gânglios simpáticos (gânglio cervical cranial) e sensitivos (gânglio trigeminal). Os resultados quantitativos mostraram que não houve diferença significativa entre os volumes dos GCCr e GT, quando comparados com o grupo controle. A respeito da avaliação qualitativa, por meio de avaliação histopatológica, foram observadas algumas alterações, como núcleos picnóticos, citoplasma anguloso, e presença marcante de células satélites nos grupos experimentais. Conclui-se que, a partir deste modelo experimental de SH, não foram encontradas diferenças quantitativas entre os grupos experimentais, tanto no GCCr, quanto no GT, porém, houve diferenças na microestrutura em ambos os gânglios, sendo que os principais achados foram corpos neurônios mortos e maior presença de células satélites.

Horners syndrome (HS) is a disease caused by the rupture or damage to the ophthalmic sympathetic chain. It occurs in many animals species including humans and its clinical signs are enophthalmus, anisocoria, ptosis and protruded third eyelid ipsilateral to the lesion. The present study aimed to evaluate possible morphoquantitative changes due to induced HS in Wistar rats. It were used 12 male rats divided into three groups of four individuals: Group I (G1): control, animals without induced HS, Group II (G2): euthanasia after two weeks from HS induction, Group III (G3): euthanasia after seven weeks from HS induction. The induction was performed surgically and the antimere was randomly chosen. The animals were anesthetised and medicated with analgesics. After the surgeries, they passed through clinical ophthalmic evaluation weekly in order to observe the clinical signs. After the determined periods for euthanasia, the biologic material was collected to quantitative evaluation (stereological evaluations about total volume, volume density and total volume of neurons) and qualitative evaluation of the sympathetic (cervical cranial ganglion) and sensitive (trigeminal ganglion) ganglia. The quantitative results showed no significance difference about CCrG and TG volumes when compared to the control group. About qualitative evaluation, made by histopathologic examination, it was observed some alterations like angulous cytoplasm, pyknotic nucleus and remarkable satellite cells presence in the experimental groups. It can be concluded that from this HS experimental model, no quantitative differences were found between the experimental groups both in the CCrG and in the GT, however, there were microstructural differences in both ganglia, with the main findings being dead neuron bodies and greater presence of satellite cells.

Selection of pacu females to hormonal induction: effect of age and of evaluation methods / Seleção de fêmeas de pacu para indução hormonal: efeito da idade e dos métodos de avaliação

In this study, we investigated the reproductive cycle and spawning season of old (OFs) and young (YFs) P. mesopotamicus females, aiming to improve the process of selection of suitable fish for induced spawning. We also evaluated the accuracy of using external body characteristics in selecting suitable females. To that, 60 OFs and 60 YFs (10 and four years old, respectively) were submitted to seven samplings during two reproductive cycles. In each sampling, females (five to 10 per treatment) were randomly chosen to evaluation of plasma concentrations of gonadal steroids and ovary composition (biopsies and stereology). Females were also submitted to an external evaluation concerning the length of abdominal thickness, the degree of the abdominal rigidity and the aspect of urogenital papilla. In spite of OFs have shown reduced concentrations of 17α- hydroxyprogesterone during the spawning season, we observed that OFs and YFs kept in captivity had similar reproductive cycles and spawning seasons (in December for both groups). We do not recommend the exclusive use of external evaluation for selecting breeders to spawning induction due to a great inaccuracy of data and lack of association with histology and ovarian biopsy, which are more accurate and reliable.(AU)

Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêmeas “jovens” (FJs) de P. mesopotamicus, visando melhorar o processo de seleção de peixes para a indução hormonal. Avaliamos também a acurácia do uso de características corporais externas para a seleção de fêmeas para indução hormonal. Para isso, 60 FVs e 60 FJs (10 e quatro anos, respectivamente) foram submetidas a sete amostragens durante dois ciclos reprodutivos. Em cada amostra, as fêmeas (cinco a 10 por tratamento) foram capturadas para avaliação das concentrações plasmáticas de esteroides gonadais e composição dos ovários (biópsias e estereologia). As fêmeas foram também submetidas à uma avaliação externa relativa à espessura abdominal, o grau de rigidez abdominal e o aspecto da papila urogenital. Exceto pela ocorrência de concentrações reduzidas de 17α-hidroxiprogesterona nas FVs, durante a época de desova, observamos que FVs e FJs, mantidas em cativeiro, apresentam ciclos reprodutivos e épocas de desova similares (em dezembro para ambos os grupos). Não recomendamos o uso exclusivo da avaliação externa para a seleção de reprodutores para indução a desova, devido a uma imprecisão dos dados e falta de associação destes com a histologia e biópsia de ovário, os quais são mais precisos e confiáveis.(AU)

Selection of pacu females to hormonal induction: effect of age and of evaluation methods / Seleção de fêmeas de pacu para indução hormonal: efeito da idade e dos métodos de avaliação

In this study, we investigated the reproductive cycle and spawning season of old (OFs) and young (YFs) P. mesopotamicus females, aiming to improve the process of selection of suitable fish for induced spawning. We also evaluated the accuracy of using external body characteristics in selecting suitable females. To that, 60 OFs and 60 YFs (10 and four years old, respectively) were submitted to seven samplings during two reproductive cycles. In each sampling, females (five to 10 per treatment) were randomly chosen to evaluation of plasma concentrations of gonadal steroids and ovary composition (biopsies and stereology). Females were also submitted to an external evaluation concerning the length of abdominal thickness, the degree of the abdominal rigidity and the aspect of urogenital papilla. In spite of OFs have shown reduced concentrations of 17α- hydroxyprogesterone during the spawning season, we observed that OFs and YFs kept in captivity had similar reproductive cycles and spawning seasons (in December for both groups). We do not recommend the exclusive use of external evaluation for selecting breeders to spawning induction due to a great inaccuracy of data and lack of association with histology and ovarian biopsy, which are more accurate and reliable.

Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêmeas “jovens” (FJs) de P. mesopotamicus, visando melhorar o processo de seleção de peixes para a indução hormonal. Avaliamos também a acurácia do uso de características corporais externas para a seleção de fêmeas para indução hormonal. Para isso, 60 FVs e 60 FJs (10 e quatro anos, respectivamente) foram submetidas a sete amostragens durante dois ciclos reprodutivos. Em cada amostra, as fêmeas (cinco a 10 por tratamento) foram capturadas para avaliação das concentrações plasmáticas de esteroides gonadais e composição dos ovários (biópsias e estereologia). As fêmeas foram também submetidas à uma avaliação externa relativa à espessura abdominal, o grau de rigidez abdominal e o aspecto da papila urogenital. Exceto pela ocorrência de concentrações reduzidas de 17α-hidroxiprogesterona nas FVs, durante a época de desova, observamos que FVs e FJs, mantidas em cativeiro, apresentam ciclos reprodutivos e épocas de desova similares (em dezembro para ambos os grupos). Não recomendamos o uso exclusivo da avaliação externa para a seleção de reprodutores para indução a desova, devido a uma imprecisão dos dados e falta de associação destes com a histologia e biópsia de ovário, os quais são mais precisos e confiáveis.

Quantitative analysis of the renal aging in rats. Stereological study

PURPOSE:To evaluate the renal function and the renal histological alterations through the stereology and morphometrics in rats submitted to the natural process of aging.METHODS:Seventy two Wistar rats, divided in six groups. Each group was sacrificed in a different age: 3, 6, 9, 12, 18 and 24 months. It was performed right nephrectomy, stereological and morphometric analysis of the renal tissue (renal volume and weight, density of volume (Vv[glom]) and numerical density (Nv[glom]) of the renal glomeruli and average glomerular volume (Vol[glom])) and also it was evaluated the renal function for the dosage of serum creatinine and urea.RESULTS:There was significant decrease of the renal function in the oldest rats. The renal volume presented gradual increase during the development of the rats with the biggest values registered in the group of animals at 12 months of age and significant progressive decrease in older animals. Vv[glom] presented statistically significant gradual reduction between the groups and the Nv[glom] also decreased significantly.CONCLUSIONS:The renal function proved to be inferior in senile rats when compared to the young rats. The morphometric and stereological analysis evidenced renal atrophy, gradual reduction of the volume density and numerical density of the renal glomeruli associated to the aging process.(AU)

Estudo morfométrico de rins em felinos domésticos (Felis catus) / Morphometric study on the kidneys of domestic cats (Felis catus)

A saúde renal tem ao longo dos últimos anos chamado atenção dos médicos veterinários, pois o comprometimento deste órgão na insuficiência renal crônica se apresenta como a maior causa de morbidade e mortalidade em felinos. O presente estudo propõe a caracterização biométrica dos rins de gatos, Felis catus, sob os aspectos macroscópicos (comprimento, altura, largura, peso e volume), mesoscópico (altura do córtex e da medula, e a sua inter-relação) e microscópica (volume glomerular), a fim de se estabelecer possíveis diferenças decorrentes de idade, sexo e simetria bilateral. Foram utilizados, rins de 30 animais da espécie Felis catus (gato deméstico), sendo 15 machos e 15 fêmeas com idade variando entre 3 meses a 15 anos, divididos em três grupos: grupo 1 (3-9 meses), grupo 2 (3-5 anos) e grupo 3 (acima de 10 anos). Os rins foram fotografados, pesados, mensurados e processados para histologia. O volume glomerular foi obtido através de estereologia. Os dados macroscópicos permitiram observar uma imparcialidade morfométrica e morfológica quando comparados os lados, independente do grupo e do sexo, sobre o comprimento, altura, largura, peso e volume dos rins. Para o sexo foi detectado diferença significante entre machos e fêmeas independente da faixa etária para comprimento, altura, peso e volume, com exceção da largura. Considerando a idade houve diferença significativa para todos os parâmetros: comprimento, altura, largura, peso e volume. Nas mensurações de córtex, medula e sua inter-relação, quando considerada a simetria bilateral, não foi detectada diferença significante entre rim direito e esquerdo. Para o sexo, não foi detectada diferença entre machos e fêmeas na mesoscopia no córtex, na medula e na inter-relação córtex/medula. Quanto à idade, a mensuração da altura do córtex e a inter-relação córtex/medula apresentou diferença significante, porém essa diferença não foi estabelecida para a medula do rim. [...] (AU)

Renal health has over the last few years called attention of veterinarians, because the chronic failure of the kidneyu is a major cause of morbidity and mortality in cats. This paper proposes biometric characterization of the kidneys from cats, Felis catus, on the macroscopic aspects (length, height, width, weight and volume), mesoscopic (height of the cortex and medulla, and their inter-relationship) and microscopic (glomerular volume) in order to establish possible differences arising from age, gender and bilateral symmetry. It where used 30 kidneys of animals from specie Felis catus (cat), 15 males and 15 females with ages between 3 months to 15 years, divided into three groups: group 1 (3-9 months), group 2 (3 - 5 years) and group 3 (above 10 years). The kidneys were photographed, weighed, measured and processed for histology. The glomerular volume was obtained by stereology. The macroscopic data allowed to observe a morphometric and morphologic impartiality when compared sides, regardless of group and sex, about the length, height, width, weight and kidney volume. Significant difference was detected between males and females regardless of age for length, height, weight and volume, except for the width. There was a significant difference for all parameters considering the age: length, height, width, weight and volume. Wasnt detected signficant diferences between left and right kdney measurements of cortex, medulla and their inter-relationship, when considered bilateral symmetry. No difference was detected between males and females in mesoscopy in the cortex, medulla and cortex interrelationship cortex/medulla. Regarding age, the measurement of the height of the cortex and the interrelationship cortex/medulla showed a significant difference, but this difference was not established to the medulla of the kidney. Significant differences were found between groups, males and females, compared to the volume occupied by the glomerulus on the parenchyma renal. [...] (AU)

MORFOLOGIA DO ENCÉFALO E NEUROTOXICOLOGIA EMBRIONÁRIA DO 2,4-D EM Tropidurus torquatus (SQUAMATA: TROPIDURIDAE)

Agrotóxicos são utilizados desde a antiguidade, porém, apenas no século passado teve início a produção sintética desses produtos, sendo o Brasil um dos maiores consumidores estas substâncias. O 2,4-D é um herbicida seletivo para ervas daninhas de folhas largas, amplamente utilizado em vários tipos de cultura. Em condições normais em animais, o 2,4-D é eliminado na urina, mas se a taxa de depuração renal for ultrapassada surgem efeitos adversos, inclusive neurotóxicos. Poucos estudos avaliam o efeito do 2,4-D sobre o desenvolvimento embrionário, sendo estes necessários pois organismos jovens são mais sensíveis que adultos. Com isso em mente visamos avaliar o efeito do 2,4-D DMA® 806 BR no desenvolvimento embrionário do encéfalo de Tropidurus torquatus. Também objetivamos descrever o encéfalo de Tropidurus torquatus como base para o estudo toxicológico. Esta espécie é abundante e por habitar lavouras pode contaminar-se com agrotóxicos naturalmente, fazendo-a um bom modelo animal. Animais adultos foram capturados e eutanaziados para a descrição macro- e microscópica do encéfalo. Fêmeas grávidas foram coletadas e mantidas em terrários até a oviposição. Os ovos foram distribuídos aleatoriamente em dois grupos, controle e 2,4-D. O substrato do grupo 2,4-D foi contaminado logo após a oviposição com uma dose correspondente à 1,5L p.c./ha, e no controle o substrato foi regado com a mesma quantidade de água destilada. De ambos os grupos foram coletados embriões nos dias 15, 30 e 60 após a oviposição. Os encéfalos dos embriões foram submetidos a inclusão em parafina para coloração com H.E. e foi estimado o volume do teto óptico através da técnica de estereologia, comparando-se os dois grupos. Foi observado um decréscimo do volume no grupo 2,4-D do dia 15, mas não foram encontradas diferenças do dia 30, sugerindo que o 2,4-D tem um leve efeito neurotóxico na dose recomendada, mas que pode ser reversível.

Pesticides have been used since antiquity, nevertheless synthetic products emerged only last century and currently Brazil is the biggest consumer of these substances. 2,4- D is a selective herbicide for broad-leaved weeds, used in several types of crops. Usually 2,4-D is eliminated in urine in animals, but when the renal clearance rate is exceeded, adverse effects, including neurotoxic ones, appear. Few studies evaluate the effect of 2,4-D on embryonic development, which is necessary because young organisms are more sensitive than adults. With this in mind we aim to evaluate the effect of 2,4-D DMA® 806 BR on the embryonic development of the brain of Tropidurus torquatus. We also aimed to describe the brain of Tropidurus torquatus as the basis for the toxicological study. This species is abundant and by inhabiting crops it can naturally contaminate itself with pesticides, making it a good animal model. Adult animals were captured and euthanized for the macro- and microscopic description of the brain. Pregnant females were collected and kept in terrariums until oviposition. Eggs were randomly distributed into two groups, control and 2,4-D. The substrate of the 2,4-D group was contaminated shortly after oviposition with a dose corresponding to 1.5L/ha, and the controls substrate was watered with the same quantity of water. Embryos were collected on days 15, 30 and 60 after oviposition. The embryos' brains were submitted to paraffin inclusion for H.E. staining and the volume of the optical tectum was estimated using stereology to compare both groups. A decrease in volume was observed in the 2,4-D group on day 15, but no differences were found on day 30, suggesting that 2,4-D has a mild neurotoxic effect at the recommended dose, which may be reversible.

POTENCIAL HEPATOPROTETOR, ANTINEOPLÁSICO E GENOPROTETOR DA GEOPRÓPOLIS PRODUZIDA POR ABELHA JANDAÍRA (Melipona subnitida D.) NO SEMIÁRIDO DO RIO GRANDE DO NORTE, BRASIL

Objetivou-se avaliar o potencial hepatoprotetor frente as injúrias induzidas por acetaminofen, a atividade antineoplásica e genoprotetora do extrato hidroetanólico da geoprópolis produzida por Melipona subnitida D., na região semiárida do Nordeste brasileiro. Foram avaliadas a composição química e atividade antioxidante das seis amostras (A1, A2, A3, A4, A5, A6) de extrato hidroetanólico da geoprópolis. Foram avaliados o potencial hepatoprotetor através das análises bioquímica da função hepática, avaliação macroscópica, histológica e estereológica dos fígados em 04 grupos experimentais (G1- controle negativo, G2- 50 mg/kg de extrato hidroetanólico da geoprópolis, G3- 50 mg/kg de extrato hidroetanólico da geoprópolis e 600 mg/kg de acetaminofen, G4- 600 mg/kg de acetaminofen) constituídos por 06 roedores da espécie Rattus norvegicus Berkenhout, 1769, linhagem Wistar, com lesão hepática aguda induzida por acetaminofen. O potencial antineoplásico foi determinado pela avaliação do efeito citotóxico do extrato hidroetanólico da geoprópolis em linhagens celulares de carcinoma hepatocelular humano (HepG2) e fibroblastos de camundongos L929. O efeito genoprotetor foi analisado em linhagem celular de fibroblastos de camundongo L929. As amostras do extrato hidroetanólico da geoprópolis apresentaram 41 compostos fenólicos, distribuídos entre fenóis e flavonoides. Relata-se pela primeira vez, em geoprópolis de meliponídeos, a presença de lignan 4- demethyldeoxypodophyllotoxin 4-O-glucopyranoside. As amostras A1, A2, A4, A5 e A6 apresentaram moderada atividade antioxidante expressas pelos valores de IC50 igual a 82,50,122,20, 98,80, 76,30 e 144,0, respectivamente, correlacionados com os elevados teores de fenóis e flavonoides totais que variaram entre 16,62 ± 3,91 a 28,46 ± 1,73 e 14,72 ± 3,75 a 32,22 ± 4,43, respectivamente. Os resultados dos níveis séricos das enzimas TGO e TGP dos ratos, indicaram que o extrato hidroetanólico da geoprópolis de M. subnitida D. não causou hepatotoxicidade. Na avaliação macroscópica dos fígados, no grupo G3 foi possível constatar coloração marrom pálida e discreta hemorragia, no G4 presença de sufusões hemorrágicas na cápsula, contornos irregulares e acentuação do padrão lobular. Na histoarquitetura dos fígados verificou no G3 focos de hepatócitos tumefeitos, vacuolizados e necrosados, no G4 degeneração e necrose difusa de hepatócitos. Na estereologia, observou aumento no percentual de hepatócitos e sinusóides, bem como, redução de colágeno no grupo G3 em relação ao G4. O extrato hidroetanólico da geoprópolis apresentou efeito citotóxico contra linhagem celular HepG2 e fibroblastos-L929, após 72 h de exposição, com valores de IC50 igual a 365,00 e 250,9 g/mL, respectivamente. Também apresentou efeito genoprotetor, com percentuais de redução de genotoxicidade de 90,74%, 73,12% e 48,18%, nas concentrações de 500, 250 e 100 l/mL do extrato, respectivamente. Conclui-se que o extrato hidroetanólico da geoprópolis de M. subnitida D. apresenta composição química variada, elevados teores de fenóis e flavonoides totais, potencial hepatoprotetor contra lesões hepáticas agudas, efeito citotóxico em células neoplásicas e atividade genoprotetora, podendo ser um produto natural promissor na quimioterapia.

The objective was to evaluate the hepatoprotective potential against injuries induced by acetaminophen, the antineoplastic and genoprotective activity of the hydroethanolic extract of the geopropolis produced by Melipona subnitida D., in the semiarid region of Northeast Brazil. The chemical composition and antioxidant activity of the six samples (A1, A2, A3, A4, A5, A6) of geopropolis hydroethanolic extract were evaluated. The hepatoprotective potential was evaluated through biochemical analysis of liver function, macroscopic, histological and stereological evaluation of the livers in 04 experimental groups (G1- negative control, G2- 50 mg/kg of hydroethanolic extract from geopropolis, G3- 50 mg/kg of hydroethanolic extract of geopropolis and 600 mg/kg of acetaminophen, G4- 600 mg/kg of acetaminophen) consisting of 06 rodents of the species Rattus norvegicus Berkenhout, 1769, Wistar strain, with acute liver damage induced by acetaminophen. The antineoplastic potential was determined by evaluating the cytotoxic effect of the hydroethanolic extract of geopropolis in human hepatocellular carcinoma (HepG2) cell lines and fibroblasts from L929 mice. The genoprotective effect was analyzed in the L929 mouse fibroblast cell line. Samples of the geopropolis hydroethanolic extract showed 41 phenolic compounds, distributed between phenols and flavonoids. The presence of 4-demethyldeoxypodophyllotoxin 4-O-glucopyranoside is reported for the first time in a meliponid geopropolis. Samples A1, A2, A4, A5 and A6 showed moderate antioxidant activity expressed by IC50 values equal to 82.50, 122.20, 98.80, 76.30 and 144.0, respectively, correlated with high levels of total phenols and flavonoids ranging from 16.62 ± 3.91 to 28.46 ± 1.73 and 14.72 ± 3.75 to 32.22 ± 4.43, respectively. The results of the serum levels of the enzymes TGO and TGP of the rats, indicated that the hydroethanolic extract of the geopropolis of M. subnitida D. did not cause hepatotoxicity. In the macroscopic evaluation of the livers, in the G3 group it was possible to observe a pale brown color and slight hemorrhage, in the G4 the presence of hemorrhagic suffusions in the capsule, irregular contours and accentuation of the lobular pattern. In the histoarchitecture of the livers, in G3 it was found foci of tumefied, vacuolized and necrotic hepatocytes, in G4 degeneration and diffuse necrosis of hepatocytes. In stereology, there was an increase in the percentage of hepatocytes and sinusoids, as well as a reduction in collagen in the G3 group compared to G4. The hydroethanolic extract of the geopropolis showed a cytotoxic effect against HepG2 cell line and L929 fibroblasts, after 72 h of exposure, with IC50 values equal to 365.00 and 250.9 g/mL, respectively. It also had a genoprotective effect, with percentages of genotoxicity reduction of 90.74%, 73.12% and 48.18%, at concentrations of 500, 250 and 100 l/mL of the extract, respectively. It is concluded that the hydroethanolic extract of the M. subnitida D. geopropolis presents varied chemical composition, high levels of total phenols and flavonoids, hepatoprotective potential against acute liver lesions, cytotoxic effect on neoplastic cells and genoprotective activity, being a promising natural product in chemotherapy.