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1.
PLoS One ; 14(12): e0226903, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31881051

RESUMO

Rearing dual-purpose chickens is a practicable approach to avoid culling one-day-old male layer chicks. The present study examined the impact of a conventional fattening diet on the liver of a novel dual-purpose chicken line (Lohmann Dual, LD) in comparison to a broiler (Ross 308) chicken line. Age-related changes of structure and lipid content of the liver were assessed. One hundred twenty and newly hatched chicks (LD = 66, Ross = 54) were kept under the same husbandry conditions and fed a commercial diet for 5 weeks for Ross and 9 weeks for LD. Six birds of each line were examined weekly. Their body weight (BW) and liver mass were recorded. Microscopic structure and ultrastructure of the liver were investigated and the liver lipid content was measured using a pre-validated method. During the study period, liver mass increased with age, while normalized liver mass decreased. Furthermore, liver mass of Ross birds was greater than that of LD birds of the same BW. Overall, no significant differences were observed in the hepatic structure or ultrastructure between the two chicken lines. The hepatic lymphatic aggregations were without fibrous capsules and their number and area increased throughout the first week, then the values began to fluctuate with age in both chicken lines. The changes in the liver lipid content of the two chicken lines were within the normal physiological range over the term of the study. The liver lipid content correlated negatively with age and body weight in both lines. It was the highest on the first day then decreased until day 7 and thereafter did not change in both chicken lines. However, given the same body weight, the Ross chickens had a 9% greater liver lipid content than LD chickens. It is concluded that there is no apparent adverse effect of a high-energy diet on the liver of LD chickens.


Assuntos
Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Gorduras/análise , Lipídeos/análise , Fígado/crescimento & desenvolvimento , Ração Animal/análise , Criação de Animais Domésticos , Animais , Peso Corporal , Fígado/química , Fígado/ultraestrutura , Masculino
2.
Toxicol Lett ; 316: 73-84, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31513886

RESUMO

In the liver microenvironment, interactions among diverse types of hepatic cells are involved in liver fibrosis. In fibrotic tissues, exosomes act as transporters in intercellular communication. Long non-coding RNAs (lncRNAs) are involved in the activation of hepatic stellate cells (HSCs), which are participants in liver fibrosis. However, the functions of exosomal lncRNAs in liver fibrosis induced by arsenite are undefined. The purposes of the present study were (a) to determine if lncRNAs secreted from human hepatic (L-02) cells exposed to arsenite are shuttled to hepatic stellate LX-2 cells and (b) to establish their effects on LX-2 cells. In mice, MALAT1 was overexpressed in the progression of liver fibrosis induced by arsenite as well as in L-02 cells exposed to arsenite. Co-cultures with arsenite-treated L-02 cells induced the activation of LX-2 cells and overexpression of MALAT1. Arsenite-treated L-02 cells transported MALAT1 into LX-2 cells. Downregulation of MALAT1, which reduced the MALAT1 levels in exosomes derived from arsenite-treated L-02 cells, inhibited the activation of LX-2 cells. Additionally, exosomal MALAT1 derived from arsenite-treated L-02 cells promoted the activation of LX-2 cells via microRNA-26b regulation of COL1A2. Furthermore, circulating exosomal MALAT1 was up-regulated in people exposed to arsenite. In sum, exosomes derived from arsenite-treated hepatic cells transferred MALAT1 to HSCs, which induced their activation. These findings support the concept that, during liver fibrosis induced by arsenite, exosomal lncRNAs are involved in cell-cell communication.


Assuntos
Arsenitos , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Exossomos/metabolismo , Células Estreladas do Fígado/metabolismo , Cirrose Hepática Experimental/metabolismo , Fígado/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Compostos de Sódio , Animais , Linhagem Celular , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Técnicas de Cocultura , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Exossomos/genética , Exossomos/ultraestrutura , Regulação da Expressão Gênica , Células Estreladas do Fígado/ultraestrutura , Humanos , Fígado/ultraestrutura , Cirrose Hepática Experimental/induzido quimicamente , Cirrose Hepática Experimental/genética , Cirrose Hepática Experimental/patologia , Masculino , Camundongos , MicroRNAs/genética , RNA Longo não Codificante/genética , Transdução de Sinais
3.
Int J Mol Sci ; 20(18)2019 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-31500117

RESUMO

Endoplasmic reticulum (ER) stress, a cellular condition caused by the accumulation of unfolded proteins inside the ER, has been recognized as a major pathological mechanism in a variety of conditions, including cancer, metabolic and neurodegenerative diseases. Trefoil factor family (TFFs) peptides are present in different epithelial organs, blood supply, neural tissues, as well as in the liver, and their deficiency has been linked to the ER function. Complete ablation of Tff3 expression is observed in steatosis, and as the most prominent change in the early phase of diabetes in multigenic mouse models of diabesity. To elucidate the role of Tff3 deficiency on different pathologically relevant pathways, we have developed a new congenic mouse model Tff3-/-/C57BL6/N from a mixed background strain (C57BL6/N /SV129) by using a speed congenics approach. Acute ER stress was evoked by tunicamycin treatment, and mice were sacrificed after 24 h. Afterwards the effect of Tff3 deficiency was evaluated with regard to the expression of relevant oxidative and ER stress genes, relevant proinflammatory cytokines/chemokines, and the global protein content. The most dramatic change was noticed at the level of inflammation-related genes, while markers for unfolded protein response were not significantly affected. Ultrastructural analysis confirmed that the size of lipid vacuoles was affected as well. Since the liver acts as an important metabolic and immunological organ, the influence of Tff3 deficiency and physiological function possibly reflects on the whole organism.


Assuntos
Estresse do Retículo Endoplasmático/genética , Fígado/metabolismo , Fator Trefoil-3/deficiência , Animais , Biomarcadores , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Expressão Gênica , Fígado/patologia , Fígado/ultraestrutura , Camundongos , Camundongos Knockout , Estresse Oxidativo/genética , Proteoma , Proteômica/métodos
4.
Mol Med Rep ; 20(3): 2101-2110, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31257518

RESUMO

Ischemia­reperfusion injury (IRI) is a notable cause of tissue damage during surgical procedures and a major risk factor in graft dysfunction in liver transplantation. Livers obtained from donors after circulatory death (DCD) are prone to IRI and toll­like receptor 4 (TLR4) serves a prominent role in the inflammatory response associated with DCD liver IRI. The present study was designed to investigate whether TAK242, a specific TLR4 inhibitor, improves hepatic IRI following a DCD graft and to investigate its underlying protective mechanisms. Male Sprague­Dawley rats were randomized into 4 groups: Control, TAK242, DCD and DCD+TAK242 groups. Rats were pretreated with TAK242 or its vehicle for 30 min, then the livers were harvested without warm ischemia (control group and TAK242 group) or with warm ischemia in situ for 30 min. The livers were stored in cold University of Wisconsin solution for 24 h and subsequently perfused for 60 min with an isolated perfused rat liver system. Rat liver injury was evaluated thereafter. When compared with the DCD group, DCD livers with TAK242 pretreatment displayed significantly improved hepatic tissue injury and less tissue necrosis (P<0.05). Compared with DCD livers, mechanistic experiments revealed that TAK242 pretreatment alleviated mitochondrial dysfunction, reduced reactive oxygen species and malondialdehyde levels and inhibited apoptosis. Additionally, TAK242 significantly inhibited the IRI­associated inflammatory response, indicated by the decreased expression of TLR4, interleukin (IL)­1ß, IL­6 and cyclooxygenase 2 at the mRNA and protein levels (P<0.05). TAK242 ameliorates DCD liver IRI via suppressing the TLR4 signaling pathway in rats. The results of the present study have revealed that TAK242 pretreatment harbors a potential benefit for liver transplantation.


Assuntos
Anti-Inflamatórios/farmacologia , Fígado/efeitos dos fármacos , Preservação de Órgãos/métodos , Traumatismo por Reperfusão/imunologia , Sulfonamidas/farmacologia , Receptor 4 Toll-Like/antagonistas & inibidores , Adenosina/farmacologia , Alopurinol/farmacologia , Animais , Glutationa/farmacologia , Insulina/farmacologia , Fígado/imunologia , Fígado/patologia , Fígado/ultraestrutura , Transplante de Fígado , Masculino , Soluções para Preservação de Órgãos/farmacologia , Rafinose/farmacologia , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/imunologia , Isquemia Quente
5.
Toxicol Mech Methods ; 29(9): 654-664, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31345115

RESUMO

Background: Application of hepatoprotectants, such as drugs or cytokines, can reduce drug-induced hepatotoxicity (DIH). Due to species-specific differences and abnormal cell polarity and drug-metabolizing enzymes (DMEs), in vivo animal models and in vitro 2D plastic dishes are not good DIH models. The aim of this study was to evaluate whether 3D re-cellularized liver is a sensitive, accurate and efficient DIH model for evaluation of hepatoprotectants. Methods: 2D plastic dishes and 3D decellular liver scaffolds were perfused with HepG2 cells or augmenter of liver regeneration (ALR)-HepG2 cells. These two cell lines were exposed to 4 µM troglitazone (TRO) or 20 µM diclofenac sodium (DIC) on day 8. DME-related genes were analyzed by quantitative reverse transcription polymerase chain reaction; morphological images were revealed by immunohistochemistry, scanning electron microscopy, transmission electron microscopy, and hematoxylin and eosin staining. Results: DME activity and cell polarity were retained and lower doses of TRO and DIC led to DIH in 3D re-cellularized liver. This DIH model reflected the protective effects and mechanism of ALR, which is one of the hepatoprotectants. ALR reduced mitochondrial damage, decreased transaminase level, and alleviated inflammation in TRO-DIH and DIC-DIH. Our re-cellularized liver lobe also showed the effect of ALR in suppressing expression of DMEs. Conclusions: Drug-induced 3D re-cellularized tissue engineering is a sensitive, accurate, and efficient DIH model for evaluation of hepatoprotectants.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Fígado/efeitos dos fármacos , Modelos Biológicos , Proteínas/genética , Engenharia Tecidual/métodos , Animais , Sobrevivência Celular/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Diclofenaco/toxicidade , Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Fígado/ultraestrutura , Substâncias Protetoras/farmacologia , Ratos Sprague-Dawley , Troglitazona/toxicidade
6.
Artif Organs ; 43(10): 1035-1041, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31211867

RESUMO

Three-dimensional tissue cultures are important models for the study of cell-cell and cell-matrix interactions, as well as, to investigate tissue repair and reconstruction pathways. Therefore, we designed a reproducible and easy to handle printable bioreactor system (Teburu), that is applicable for different approaches of pathway investigation and targeted tissue repair using human tissue slices as a three-dimensional cell culture model. Here, we definitively describe Teburu as a controlled environment to reseed a 500-µm thick decellularized human liver slice using human mesenchymal stroma cells. During a cultivation period of eight days, Teburu, as a semi-open and low consumption system, was capable to maintain steady pH and oxygenation levels. Its combination with additional modules delivers an applicability for a wide range of tissue engineering approaches under optimal culture conditions.


Assuntos
Bioimpressão , Reatores Biológicos , Impressão Tridimensional , Técnicas de Cultura de Tecidos/instrumentação , Desenho de Equipamento , Humanos , Fígado/química , Fígado/citologia , Fígado/ultraestrutura , Engenharia Tecidual/instrumentação , Tecidos Suporte/química
7.
Biomed Pharmacother ; 117: 109073, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31212129

RESUMO

Dendrobium nobile Lindl. alkaloids (DNLA), the active ingredients of Dendrobium, has been shown to possess anti-oxidative effects. The nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant signaling pathway plays a critical role in the cellular response to oxidative stress. Oxidative damage has been implicated in the mechanism of various hepatotoxins induced liver injury. The present study aimed to examine the protective effects of DNLA on CCl4-induced acute liver injury, and to explore the role of the Nrf2 pathway in the protective action of DNLA. Wild-type (WT) and Nrf2-knockout (Nrf2-/-) mice were administrated with DNLA (20 mg/kg/day, ig) for 7 days, and then challenged with CCl4 (20 µL/kg, ip). In WT mice, DNLA reduced CCl4 induced liver injury, as evidenced by the reduction in the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), attenuation of malondialdehyde (MDA) production, and improved ultrastructural morphology in hepatocytes. However, the protective effect was diminished in Nrf2-/- mice, indicating an essential role of Nrf2 in DNLA-mediated protection over CCl4 liver injury. Furthermore, it was found that DNLA enhanced Nrf2 expression and nuclear accumulation and increased the expression of Nrf2 regulated downstream proteins. These results demonstrate that DNLA protects mice from CCl4 induced liver injury, probably through the activation of the Nrf2 signaling pathway.


Assuntos
Alcaloides/farmacologia , Dendrobium/química , Fígado/lesões , Fator 2 Relacionado a NF-E2/metabolismo , Substâncias Protetoras/farmacologia , Transdução de Sinais , Doença Aguda , Animais , Tetracloreto de Carbono , Fígado/efeitos dos fármacos , Fígado/patologia , Fígado/ultraestrutura , Masculino , Malondialdeído/metabolismo , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/deficiência
8.
Aquat Toxicol ; 213: 105195, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31203167

RESUMO

Titanium dioxide nanoparticles (n-TiO2) are among the man-made nanomaterials that are predicted to be found at high concentrations in the aquatic environment. There, they likely co-exist with other chemical pollutants. Thus, n-TiO2 and other chemical pollutants can be taken up together or accumulate independently from each other in prey organisms of fish. This can lead to dietary exposure of fish to n-TiO2-chemical pollutant mixtures. In this study, we examine if simultaneous dietary exposure to n-TiO2 and 3,3',4,4'-Tetrachlorobiphenyl (PCB77) -used as a model compound for persistent organic pollutants with dioxin-like properties- can influence the uptake and toxicological response elicited by the respective other substance. Juvenile brown trout (Salmo trutta) were fed custom-made food pellets containing n-TiO2, PCB77 or n-TiO2+PCB77 mixtures for 15 days. Ti and PCB77 concentrations in the liver were measured by ICP-MS and GC-MS, respectively. Besides, n-TiO2 uptake was assessed using TEM. Combination effects on endpoints specific for PCB77 (i.e., cytochrome P450 1A (CYP1A) induction) and endpoints shared by both PCB77 and n-TiO2 (i.e., oxidative stress-related parameters) were measured in intestine and liver using RT-qPCR and enzyme activity assays. The results show that genes encoding for proteins/enzymes essential for tight junction function (zo-1) and ROS elimination (sod-1) were significantly upregulated in the intestine of fish exposed to n-TiO2 and PCB77 mixtures, but not in the single-substance treatments. Besides, n-TiO2 had a potentiating effect on PCB77-induced CYP1A and glutathione reductase (GR) expression/enzyme activity in the liver. This study shows that simultaneous dietary exposure to nanomaterials and traditional environmental pollutants might result in effects that are larger than observed for the substances alone, but that understanding the mechanistic basis of such effects remains challenging.


Assuntos
Dieta , Exposição Ambiental , Nanopartículas/toxicidade , Bifenilos Policlorados/toxicidade , Titânio/toxicidade , Truta/fisiologia , Animais , Biomarcadores/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Comportamento Alimentar , Regulação da Expressão Gênica/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/ultraestrutura , Metalotioneína/metabolismo , Nanopartículas/ultraestrutura , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Poluentes Químicos da Água/toxicidade
9.
Int. j. morphol ; 37(2): 706-711, June 2019. graf
Artigo em Inglês | LILACS | ID: biblio-1002281

RESUMO

A serous membrane covering the liver and the hepatic parenchyma, consists of hepatocytes, arteries, veins, hepatic sinusoids and biliary ductuli. There are erythrocytes, thrombocytes, melanin particles and Kupffer cell in the hepatic sinusoids and the blood vessels. The gall bladder wall consists of a mucous layer a muscle layer and a serous layer. The bottom of the epithelium abounds with round or oval secretory. In liver, immunohistochemistry results show that AQP1 have intense reaction in hepatic lobule, Kupffer cells (Macrophagocytus stellatus), hepatocytes, portal tract, blood islands, vein and artery, but almost no reaction of AQP2 was detected. In gallbladder, mucous epithelium, endothelial cells from vein and artery all have strong AQP1 expression, AQP2 showed minor diffused positive reaction in gallbladder, which suggesting that AQP1 may have the main role in the absorption and transportation of fluid in hepatobiliary system of Qinghai Lizard.


Una membrana serosa cubre el hígado y el parénquima hepático el cual está formado por hepatocitos, arterias, venas, sinusoides hepáticos y conductos biliares. Se encuentran eritrocitos, trombocitos, partículas de melanina y células de Kupffer en los sinusoides hepáticos y en los vasos sanguíneos. La pared de la vesícula biliar presenta tres capas: mucosa, muscular y serosa. En el hígado, la inmunohistoquímica mostró que AQP1 tiene una reacción intensa en el lóbulo hepático, células de Kupffer, hepatocitos, tracto portal e islotes sanguíneos. En venas y arterias, no se detectó reacción alguna de AQP2. En la vesícula biliar, el epitelio mucoso, las células endoteliales venosas y arteriales tuvieron una importante expresión de AQP1, sin embargo, AQP2 mostró una reacción positiva difusa menor, lo que sugiere que la AQP1 podría tener una función principal en la absorción y transporte de líquido en el sistema hepatobiliar del Lagarto Qinghai.


Assuntos
Animais , Aquaporinas/metabolismo , Vesícula Biliar/metabolismo , Fígado/metabolismo , Lagartos , Imuno-Histoquímica , Aquaporina 1/metabolismo , Aquaporina 2/metabolismo , Vesícula Biliar/ultraestrutura , Fígado/ultraestrutura
10.
N Biotechnol ; 52: 69-83, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31082574

RESUMO

Pre-analytical factors can greatly influence the outcome of molecular analyses in medical diagnostics and research. This also applies to in situ staining techniques such as immunohistochemistry (IHC), where different types of tissue fixation methods lead to different modifications of proteins and thus can affect differently the detection by antibodies. For formalin-fixed paraffin-embedded (FFPE) tissue, antigen retrieval is applied in order to reverse the negative effects of formalin and re-establish immunoreactivity. Most antibodies and protocols used in IHC are optimized for FFPE tissue, but not for paraffin-embedded tissue treated with other fixatives such as non-crosslinking fixatives. We report results from systematic studies on distinct pre-analytical conditions in IHC, immunofluorescence and electron microscopy. Parameters investigated are the impact of crosslinking and non-crosslinking fixatives (comparing formalin and PAXgene Tissue fixation) on whole tissue, subcellular structures and organelles, as well as on ultrastructure. The results generated show that minor changes in antigen retrieval conditions may have a major impact on IHC results and that protocols optimized for crosslinking fixatives may not be used for other fixatives without re-validation. Key antigen retrieval parameters such as buffers with different pH and duration of microwave treatment must be tested systematically for each antibody and fixation protocol.


Assuntos
Antígenos/metabolismo , Reagentes para Ligações Cruzadas/química , Fixadores/química , Imuno-Histoquímica/métodos , Animais , Neoplasias da Mama/patologia , Neoplasias do Colo/patologia , Feminino , Células Hep G2 , Humanos , Fígado/metabolismo , Fígado/ultraestrutura , Camundongos , Proteínas de Neoplasias/metabolismo , Coloração e Rotulagem , Proteína Supressora de Tumor p53/metabolismo
11.
Braz J Med Biol Res ; 52(6): e7628, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31116255

RESUMO

This study aimed to explore the influence of gut microbiota alterations induced by Linderae radix ethanol extract (LREE) on alcoholic liver disease (ALD) in rats and to study the anti-inflammatory effect of LREE on ALD through the lipopolysaccharide (LPS) toll-like receptor 4 (TLR4)-nuclear factor kappa B (NF-κB) pathway. ALD rat models were established by intragastric liquor [50% (v/v) ethanol] administration at 10 mL/kg body weight for 20 days. Rats were divided into six groups: normal group (no treatment), model group (ALD rats), Essentiale group (ALD rats fed with Essentiale, 137 mg/kg), and LREE high/moderate/low dose groups (ALD rats fed with 4, 2, or 1 g LREE/kg). NF-κB and LPS levels were evaluated. Liver pathological changes and intestinal ultrastructure were examined by hematoxylin and eosin staining and transmission electron microscopy. The gut microbiota composition was evaluated by 16S rDNA sequencing. Expression levels of TLR4 and CD68 in liver tissue, and occludin and claudin-1 in intestinal tissue were measured. LREE treatment significantly reduced NF-κB and LPS levels, improved liver pathological changes, and ameliorated intestinal ultrastructure injury. Meanwhile, LREE-fed groups showed a higher abundance of Firmicutes and a lower abundance of Bacteroidetes than the rats in the model group. Administration of LREE suppressed TLR4 overexpression and promoted the expression of occludin and claudin-1 in intestine tissue. Thus, LREE could partly ameliorate microflora dysbiosis, suppress the inflammatory response, and attenuate liver injury in ALD rats. The protective effect of LREE might be related to the LPS-TLR4-NF-κB pathway.


Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Inflamação/prevenção & controle , Lindera/química , Hepatopatias Alcoólicas/prevenção & controle , Fígado/ultraestrutura , Extratos Vegetais/farmacologia , Animais , Citocinas/sangue , Modelos Animais de Doenças , Lipopolissacarídeos/sangue , Hepatopatias Alcoólicas/diagnóstico por imagem , Masculino , Raízes de Plantas/química , Proteínas Serina-Treonina Quinases/sangue , Ratos , Ratos Sprague-Dawley , Receptor 4 Toll-Like/sangue
12.
Acta Histochem ; 121(5): 563-574, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31072619

RESUMO

The accidental spilling of petroleum oils into natural water resources expose fishes in the effluent area to serious problems.. Oreochromis niloticus were used in the current study as a model to investigate the toxicity of used engine oil and to evaluate the protective role of vitamin C against this toxicity. The oil concentration used in this study was previously determined to be 0.25 ml/l by 96 h-LC50. After 21 days of engine oil exposure, haematological and biochemical analyses revealed significant reduction in RBCs counts, haemoglobin concentrations and total proteins. However, ALT, AST and glucose levels were significantly increased by the end of the experiment indicating the damaging effects of the oil on fish tissues. Oxidative stress biomarkers were also measured; liver CAT activity was significantly decreased in the oil exposed group compared to control group, while MDA levels were significantly elevated. Histopathological examination showed the presence of several alterations in hepatic and branchial tissues in exposed group compared to the control group. Significant elevations in CYP1 A1 mRNA expression levels in hepatic tissue were also detected in the group exposed to used engine oil compared to the control group. However, supplementation of fishexposed to used engine oil with vitamin Csignificantly enhance the biochemical, oxidative and histological parameters.


Assuntos
Ácido Ascórbico/farmacologia , Ciclídeos , Brânquias/efeitos dos fármacos , Fígado/efeitos dos fármacos , Petróleo/toxicidade , Animais , Análise Química do Sangue , Ciclídeos/sangue , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Brânquias/patologia , Brânquias/ultraestrutura , Nível de Saúde , Histocitoquímica , Fígado/patologia , Fígado/ultraestrutura , Microscopia Eletrônica de Transmissão , Estresse Oxidativo/efeitos dos fármacos
13.
Nat Protoc ; 14(6): 1756-1771, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31053799

RESUMO

In vitro 3D organoid systems have revolutionized the modeling of organ development and diseases in a dish. Fluorescence microscopy has contributed to the characterization of the cellular composition of organoids and demonstrated organoids' phenotypic resemblance to their original tissues. Here, we provide a detailed protocol for performing high-resolution 3D imaging of entire organoids harboring fluorescence reporters and upon immunolabeling. This method is applicable to a wide range of organoids of differing origins and of various sizes and shapes. We have successfully used it on human airway, colon, kidney, liver and breast tumor organoids, as well as on mouse mammary gland organoids. It includes a simple clearing method utilizing a homemade fructose-glycerol clearing agent that captures 3D organoids in full and enables marker quantification on a cell-by-cell basis. Sample preparation has been optimized for 3D imaging by confocal, super-resolution confocal, multiphoton and light-sheet microscopy. From organoid harvest to image analysis, the protocol takes 3 d.


Assuntos
Imagem Tridimensional/métodos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Imagem Óptica/métodos , Organoides/ultraestrutura , Fixação de Tecidos/métodos , Animais , Mama/ultraestrutura , Colo/ultraestrutura , Feminino , Humanos , Imuno-Histoquímica/métodos , Rim/ultraestrutura , Fígado/ultraestrutura , Camundongos
14.
Acta Histochem ; 121(5): 575-583, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31078256

RESUMO

Almost all transplanted solid organs are exposed to some degree of ischemia-reperfusion (IR) damage. It is interesting to know that this IR damage affects various remote tissues including the liver and resulted in serious adverse effects. Liver injury triggers different responses of liver tissue especially Kupffer cells (KCs). The goal of this current study is to assess the biochemical and morphological changes of hepatic KCs after the induction of renal ischemia-reperfusion (RIR) and point out their role in remote liver injury after RIR. Sixteen male Sprague-Dawley rats were randomly divided into two equal groups: Group I; sham group. Group II; renal ischemia reperfusion (IR) group in which rats were exposed to renal ischemia for 45 min followed by renal reperfusion for 48 h. Three rats from each group were subjected to charcoal injection to evaluate KCs activity. Specimens of rat liver from each group were obtained and processed for biochemical, light microscopic and ultramicroscopic examination. The current results showed elevated serum levels of AST and ALT. The liver HGF-α protein expression increased in IR group compared to the sham group. In IR group, numerous charcoal labeled KCs were observed mainly localized around the central vein. Scanning electron micrographs showed complex primary and secondary foot process of the KCs. Ultrastructural study showed KCs with multiple cytoplasmic vacuoles, lysosomes and mitochondria, rough endoplasmic reticulum and ribosomes. Immuno-histochemical study showed more tumor necrosis factor-α (TNF-α) expression in KCs than the sham group. These results collectively demonstrated that renal IR produced biochemical and morphological changes in the liver KCs and theses cells might have a role in the remote liver injury after renal IR. This might be one of the mechanisms through which RIR affects the liver.


Assuntos
Nefropatias/patologia , Macrófagos do Fígado/patologia , Traumatismo por Reperfusão/patologia , Animais , Imuno-Histoquímica , Nefropatias/metabolismo , Macrófagos do Fígado/metabolismo , Macrófagos do Fígado/ultraestrutura , Fígado/metabolismo , Fígado/fisiopatologia , Fígado/ultraestrutura , Masculino , Microscopia Eletrônica , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
15.
Ann Anat ; 224: 124-132, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31100343

RESUMO

BACKGROUND: Autoimmune hepatitis (AIH) is an immune-mediated inflammation of the liver characterized by disorganized hepatic parenchyma and inflammatory cell infiltration. Although the increased incidence of AIH, the development of novel therapeutic strategies are impeded by the poor understanding of the accompanied detailed immunopathogenic changes. CD4+ T cells are key mediators of inflammatory cell infiltration in initial phases of liver injuries like AIH. The distribution of CD4+ cells and the histopathological changes accompanying Con A-induced AIH were investigated together with the postulated protective effect of Amygdalin (Amg.). MATERIALS AND METHODS: 30 adult male mice were divided into three groups; control, AIH and AIH-Amg. groups. AIH was induced by a single intravenous injection of Concanavalin A (Con A) (15 mg/kg). The AIH-Amg. group received Amg. 5 mg/kg intraperitoneally once a week for three weeks. Blood samples were examined for ALT and AST. MDA, SOD, and GSH were determined in hepatic homogenates. Liver section stained with hematoxylin and eosin, Masson trichrome and CD4+ immune stain were examined by light and electron microscopy. RESULTS: AIH group showed a significant increase in levels of ALT, AST and MDA and a significant decline in SOD and GSH compared to the controls. The liver tissue showed distorted hepatic architecture with intercellular hemorrhage, necrosis, and inflammatory cell infiltration. The area percent of CD4+ immune staining was significantly increased. Electron microscopic examination showed massive cellular degenerative changes. Amg. pretreatment in AIH-Amg. group significantly reversed these changes. CONCLUSION: AIH induced CD4+ cells infiltration in the liver with subsequent liver tissue damage. Amg. pretreatment inhibited CD4+ cell infiltration and protected the liver tissue. This finding suggests that Amg. could be a therapeutic agent in the management of AIH.


Assuntos
Amigdalina/administração & dosagem , Linfócitos T CD4-Positivos/efeitos dos fármacos , Hepatite Autoimune/tratamento farmacológico , Fígado/imunologia , Fígado/patologia , Alanina Transaminase/sangue , Análise de Variância , Animais , Aspartato Aminotransferases/sangue , Concanavalina A/efeitos adversos , Glutationa/metabolismo , Hepatite Autoimune/etiologia , Hepatite Autoimune/imunologia , Hepatite Autoimune/prevenção & controle , Imuno-Histoquímica , Fígado/efeitos dos fármacos , Fígado/ultraestrutura , Masculino , Malondialdeído/metabolismo , Camundongos , Mitógenos/efeitos adversos , Estresse Oxidativo , Superóxido Dismutase/metabolismo
16.
Chemosphere ; 228: 685-693, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31063915

RESUMO

Di-(2-ethylhexyl) phthalate (DEHP) is a widespread environmental toxicant that severely impacts agricultural production and animal and human health. Nevertheless, DEHP-induced hepatotoxicity at the molecular level in quail remains unexplored. The heat shock response (HSR), involving heat shock proteins (HSPs) and heat shock transcription factors (HSFs), is a highly conserved molecular response that is triggered by stressors, especially exposure to toxicants. To explore the DEHP-induced hepatotoxicity that occurs via regulation of HSR in birds, female quail were dosed with DEHP by oral gavage (0, 250, 500 and 1000 mg/kg) for 45 days. Based on histopathological analysis, the livers of the DEHP-treated groups exhibited structural alterations of hepatocytes, including mitochondrial swelling, derangement of hepatic plates, inflammatory cell infiltration and adipose degeneration. Ultrastructural evaluation of the livers of DEHP-treated quail revealed swollen mitochondria, partial disappearance of mitochondrial membranes and cristae, nuclear chromatin margination and nuclear condensation. The expression of HSF1 and HSF3 significantly decreased after DEHP exposure. The levels of HSPs (HSP10, HSP25, HSP27, HSP40, HSP47, HSP60, HSP70 and HSP90) were significantly downregulated in the livers of DEHP-treated quail. In this study, we concluded that DEHP exposure resulted in liver function damage and hepatotoxicity by reducing the expression of HSFs and HSPs in quail liver, which inhibited the protective effect of the HSR signaling pathway.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/etiologia , Coturnix/fisiologia , Dietilexilftalato/toxicidade , Resposta ao Choque Térmico/efeitos dos fármacos , Fígado/efeitos dos fármacos , Animais , Ecotoxicologia/métodos , Poluentes Ambientais/toxicidade , Feminino , Proteínas de Choque Térmico/metabolismo , Humanos , Fígado/patologia , Fígado/ultraestrutura , Transdução de Sinais/efeitos dos fármacos
17.
Environ Toxicol ; 34(7): 878-885, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31037826

RESUMO

Perfluorooctanoic acid (PFOA) is an octanoic acid and is found in wildlife and humans. We have investigated mitochondrial toxicity in isolated mitochondria from, placenta, brain, liver, and heart after oral exposure with PFOA in mice during gestational days (7-15). Histopathological examination and mitochondrial toxicity parameters were assayed. Results indicated that PFOA decreased the weight of the fetus and placenta, the length of the fetus and the diameter of the placenta, dead fetuses and dead macerated fetuses in treated mice with 25 mg/kg. Histopathological examination showed that PFOA induced pathological abnormalities in liver, brain, heart, and placenta. Also, PFOA induced mitochondria toxicity in brain, liver, heart of mouse fetus. Our results indicate that PFOA up to 20 mg/kg exposure adversely affect embryofetal/developmental because for mitochondria dysfunction. These results suggested that mitochondrial dysfunction induced by PFOA in liver, heart, and brain lead to developmental toxicity and abnormality in tissues.


Assuntos
Aborto Espontâneo/induzido quimicamente , Caprilatos/toxicidade , Desenvolvimento Fetal/efeitos dos fármacos , Feto/efeitos dos fármacos , Fluorcarbonetos/toxicidade , Exposição Materna/efeitos adversos , Mitocôndrias/efeitos dos fármacos , Anormalidades Induzidas por Medicamentos/patologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/ultraestrutura , Feminino , Feto/metabolismo , Feto/patologia , Coração/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/ultraestrutura , Masculino , Camundongos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Mitocôndrias Cardíacas/efeitos dos fármacos , Mitocôndrias Cardíacas/fisiologia , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/fisiologia , Miocárdio/ultraestrutura , Gravidez
18.
Artigo em Inglês | MEDLINE | ID: mdl-31028934

RESUMO

Here we used RNA-Seq to explore the transcriptomic response and specific involvement of hepatic mRNA of juvenile Oreochromis niloticus (GIFT) as a result of dietary resveratrol supplementation (0.05 g/kg RES). More than 24,513,018 clean reads were reference genome guided assembly into 23,417 unigenes. 12,596 unigenes (29.64%) were annotated to GO database. There were 5, 179 and 1526 genes significantly differentially expressed genes at 15, 30 and 45 d respectively, and 8 KEGG pathways were enriched associated with this immune response. Hyperemia and compressed hepatic sinusoid, fibrosis of liver cell and abnormal hepatic epidermal cell revealed by H&E and SEM analysis respectively. Genes related with cytokine production (il12rb2, scfr), immune system (ig8l, hlfl, cd226, prf1l), autophagy regulation (atg4b), foxo signaling (ccnb2), steroid hormone biosynthesis (cyp3a40), fatty acid metabolism (scd1), metabolism (cacna1b) have been significantly decreased, while genes associated with such pathways above (leap-2, prdx4, mb, homer1, mif, sat1, cytbc1_8) and the pathway of protein processing in endoplasmic reticulum (cne1, tram1) have been significantly increased. These findings suggested RES activated some immune and biological process-related genes to enhance GIFT's innate immunity. It also suggested high concentration addition or long-time administration may bring negative effect in tilapia liver.


Assuntos
Ciclídeos/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Fígado/efeitos dos fármacos , Resveratrol/farmacologia , Animais , Animais Geneticamente Modificados , Aquicultura , Suplementos Nutricionais , Perfilação da Expressão Gênica , Imunidade Inata/efeitos dos fármacos , Imunidade Inata/genética , Fígado/fisiologia , Fígado/ultraestrutura , Microscopia Eletrônica de Varredura , Anotação de Sequência Molecular , Reprodutibilidade dos Testes
19.
Avian Pathol ; 48(4): 285-287, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30942612

RESUMO

Campylobacter hepaticus was recently identified as the aetiological agent of Spotty Liver Disease (SLD). SLD causes significant health and productivity losses in the Australian egg industry and the disease is present in other countries. Following the isolation and characterization of C. hepaticus, molecular tools and refined culturing methods have been developed to identify the pathogen. It is suspected that the application of these tools will lead to identification of the pathogen in many poultry production systems throughout the world. As C. hepaticus has only recently been identified, little is known about the mechanisms of pathogenesis and, hence, new research needs to be directed towards understanding SLD epidemiology and C. hepaticus virulence mechanisms to inform efforts to develop intervention strategies.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter , Galinhas , Hepatopatias/veterinária , Doenças das Aves Domésticas/microbiologia , Animais , Campylobacter/ultraestrutura , Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/terapia , Fígado/microbiologia , Fígado/patologia , Fígado/ultraestrutura , Hepatopatias/microbiologia , Hepatopatias/terapia , Microscopia Eletrônica de Transmissão/veterinária , Doenças das Aves Domésticas/terapia
20.
Curr Pharm Biotechnol ; 20(6): 465-475, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30961481

RESUMO

BACKGROUND: Zinc oxide nanoparticles (ZnO NPs) are increasingly utilized in both industrial and medical applications. Therefore, the study was aimed to investigate the effect of green nanoparticle complex (green tea extract/zinc oxide nanoparticles complex, GTE/ZnO NPs) on oxidative stress induced by monosodium glutamate (MSG) on the liver of rats. METHODS: Wistar male rats (n=64) weighing between 200-250 g were divided randomly into eight groups: control group was given physiological saline (1 mg/kg), two groups were treated with two different doses of MSG (MSG-LD, MSG-HD; 6 and 17.5 mg/Kg, respectively), GTE was given 1 mg/mL, 5th group was treated with ZnO NPs and 6th group was treated with GTE/ZnO NPs complex while, 7th and 8th groups were treated with MSG-LD + GTE/ZnO NPs complex and MSG-HD + GTE/ZnO NPs complex, respectively. All substances were given orally for 30 consecutive days. At the end of the study, the liver was homogenized for measurement of the oxidative stress status and anti-inflammatory biomarkers as well as histological and transmission alternations. RESULTS: Results showed that the antioxidant enzymes activity and glutathione level were significantly decreased in MSG groups than control in a dose-dependent manner. Conversely, the malondialdehyde and inflammatory cytokines levels were significantly increased in MSG groups than the control group. The liver indicated no evidence of alteration in oxidative status, anti-inflammatory and morphological parameters in GTE, ZnO NPs and GTE/ZnO NPs complex groups. CONCLUSION: In conclusion, MSG at both doses caused oxidative stress and inflammation on liver after 28 days of exposure that supported histological analysis and transmission view of hepatic parenchyma. GTE/ZnO NPs act as partial hepato-protective against MSG.


Assuntos
Camellia sinensis/química , Fígado/efeitos dos fármacos , Nanopartículas Metálicas/química , Extratos Vegetais/farmacologia , Glutamato de Sódio/toxicidade , Óxido de Zinco/farmacologia , Animais , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Citocinas/metabolismo , Fígado/enzimologia , Fígado/ultraestrutura , Masculino , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/imunologia , Ratos , Ratos Wistar
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