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1.
Acta Cir Bras ; 34(6): e201900606, 2019 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-31432997

RESUMO

PURPOSE: To investigate the effects of pine needle extract (PNE) on the expression of proliferating cell nuclear antigen (PCNA) and Ki-67 during liver regeneration induced by 70% partial hepatectomy (PH) in rat. METHODS: Forty-eight male rats (SD, 7 weeks) had surgery (70% PH). They were randomly divided into two groups. PH + PNE group was only provided PNE diluted in water (10%) for drinking and PH group was provided water from 5 days before surgery to the time of sacrifice. PNE was made by pressing and filtering. Animals were sacrificed at 12h, 24h, 36h, 60h, 84h, 168h after PH, respectively. The expressions of PCNA and Ki-67 were determined as proliferation indices. RESULTS: Immunohistochemistry turned out to increase the expression of PCNA and Ki-67. PCNA expression of PH+PNE group increased up to twice of that of PH group. Western blot also seemed to increase the PCNA expression. These results indicated the promotion of cell proliferation in liver tissue and hepatic regeneration. CONCLUSIONS: Pine needle extract stimulates the expression of some mitotic proteins during liver regeneration induced by 70% PH in rats. It suggests that administration of pine needle extract could accelerate the liver regeneration after partial hepatectomy.


Assuntos
Hepatectomia/métodos , Antígeno Ki-67/efeitos dos fármacos , Regeneração Hepática/efeitos dos fármacos , Pinus/química , Extratos Vegetais/farmacologia , Antígeno Nuclear de Célula em Proliferação/efeitos dos fármacos , Animais , Proliferação de Células , Antígeno Ki-67/metabolismo , Masculino , Índice Mitótico , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de Tempo
2.
Int J Nanomedicine ; 14: 4723-4739, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31308655

RESUMO

Background: Much consideration has been paid to the toxicological assessment of nanoparticles prior to clinical and biological applications. While in vitro studies have been expanding continually, in vivo investigations of nanoparticles have not developed a cohesive structure. This study aimed to assess the acute toxicity of different concentrations of chitosan-coated silver nanoparticles (Ch-AgNPs) in main organs, including liver, kidneys, and spleen. Materials and methods: Twenty-eight male albino rats were used and divided into 4 groups (n=7). Group 1 was kept as a negative control group. Groups 2, 3, and 4 were treated intraperitoneally with Ch-AgNPs each day for 14 days at doses of 50, 25, and 10 mg/kg body weight (bwt) respectively. Histopathological, morphometric and immunohistochemical studies were performed as well as oxidative stress evaluations, and specific functional examinations for each organ were elucidated. Results: It was revealed that Ch-AgNPs induced dose-dependent toxicity, and the repeated dosing of rats with 50 mg/kg Ch-AgNPs induced severe toxicities. Histopathological examination showed congestion, hemorrhage, cellular degeneration, apoptosis and necrosis in hepatic and renal tissue as well as lymphocytic depletion with increasing tangible macrophages in the spleen. The highest levels of malondialdehyde, alanine aminotransferase, aspartate aminotransferase (MDA, ALT, AST) and the lowest levels of reduced glutathione, immunoglobulin G, M and total protein (GSH, IgG, IgM, TP) were observed in this group. On the other hand, repeated dosing with 25 mg/kg induced mild to moderate disturbance in the previous parameters, while there was no significant difference in results of pathological examination and biochemical tests between the control group and those treated with 10 mg/kg bwt Ch-AgNPs. Conclusion: Chitosan-coated silver nanoparticles induce dose-dependent adverse effects on rats.


Assuntos
Quitosana/química , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Nitrogênio da Ureia Sanguínea , Caspase 3/metabolismo , Creatinina/sangue , Glutationa/metabolismo , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Malondialdeído/metabolismo , Nanopartículas Metálicas/ultraestrutura , Estresse Oxidativo/efeitos dos fármacos , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos Wistar , Baço/efeitos dos fármacos , Baço/patologia
3.
Nat Commun ; 10(1): 2420, 2019 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-31160570

RESUMO

Replication-Factor-C (RFC) and RFC-like complexes (RLCs) mediate chromatin engagement of the proliferating cell nuclear antigen (PCNA). It remains controversial how RFC and RLCs cooperate to regulate PCNA loading and unloading. Here, we show the distinct PCNA loading or unloading activity of each clamp loader. ATAD5-RLC possesses the potent PCNA unloading activity. ATPase motif and collar domain of ATAD5 are crucial for the unloading activity. DNA structures did not affect PCNA unloading activity of ATAD5-RLC. ATAD5-RLC could unload ubiquitinated PCNA. Through single molecule measurements, we reveal that ATAD5-RLC unloaded PCNA through one intermediate state before ATP hydrolysis. RFC loaded PCNA through two intermediate states on DNA, separated by ATP hydrolysis. Replication proteins such as Fen1 could inhibit the PCNA unloading activity of Elg1-RLC, a yeast homolog of ATAD5-RLC in vitro. Our findings provide molecular insights into how PCNA is released from chromatin to finalize DNA replication/repair.


Assuntos
ATPases Associadas a Diversas Atividades Celulares/metabolismo , Replicação do DNA , Proteínas de Ligação a DNA/metabolismo , DNA/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteína de Replicação C/metabolismo , Adenosina Trifosfatases , Trifosfato de Adenosina/metabolismo , Proteínas de Transporte/metabolismo , Cromatina/metabolismo , Endonucleases Flap/metabolismo , Humanos , Hidrólise , Saccharomyces cerevisiae , Proteínas de Saccharomyces cerevisiae/metabolismo
4.
Int. j. morphol ; 37(2): 515-521, June 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1002253

RESUMO

SUMMARY: Reproductive dysfunction is a complication for many diseases and toxins. Its early diagnosis and treatment are immensely important. Here the morphological histoarchitecture changes in early testicular and cauda toxicity before and after treatment with angiotensin receptor blockers were evaluated. Low-grade testicular damage was induced using thioacetamide (TAA, 50 mg/kg/day) intraperitoneally for two weeks in rats. The rats were randomly divided into four groups (n = 8) treated daily orally for three weeks as follows: Normal control (distilled water), TAA (positive control), TAA+candesartan (0.2 mg/kg) and TAA+losartan (7.5 mg/kg). Serum testosterone and testicular malondialdehyde and glutathione were measured. The changes in histoarchitecture of testis and cauda epididymis were evaluated by hematoxylin and eosin for general structure, Masson's trichrome for collagen, periodic acid Schiff for basement membrane, and caspase-3 and proliferating cell nuclear antigen (PCNA) for immunohistochemical analysis. The TAA-rats showed decreases of serum testosterone and testicular glutathione, increases in testicular malondialdehyde, degenerative changes and apoptosis in germ cells, thickening of tubular basal lamina and increases in expression of caspase 3, and decreases in expression of PCNA. The ARBs (candesartan and losartan) significantly reversed these changes with non-significant differences in-between. Treatment with ARBs (candesartan and losartan) significantly reversed TAA-induced low-grade testicular and cauda toxicity in rats. This could be potentially useful for early treatment of male patients with occupational toxicant-induced reproductive dysfunction especially if they are using ARBs for other comorbidities.


RESUMEN: La disfunción reproductiva es una complicación por muchas enfermedades y toxinas. Su diagnóstico y tratamiento tempranos son inmensamente importantes. Aquí se evaluaron los cambios morfológicos en la histoarquitectura en la toxicidad precoz testicular y cauda antes y después del tratamiento con bloqueadores de receptores de angiotensina. Se indujo daño testicular de bajo grado usando tioacetamida (TAA, 50 mg / kg / día) por vía intraperitoneal durante dos semanas en ratas. Las ratas se dividieron aleatoriamente en cuatro grupos (n = 8) tratados diariamente por vía oral durante tres semanas de la siguiente manera: control normal (agua destilada), TAA (control positivo), TAA + candesartan (0,2 mg / kg) y TAA + losartán (7,5 mg / kg). Se midieron la testosterona sérica, el malondialdehído testicular y el glutatión. Los cambios en la histoarquitectura de los testículos y la epidermis de la cauda se evaluaron mediante Hematoxilina y Eosina para determinar la estructura general, con tricrómicro de Masson para el colágeno, ácido periódico de Schiff para la membrana basal y la caspasa-3 y el antígeno nuclear de células proliferantes (PCNA) para análisis inmunohistoquímico. Las ratas TAA mostraron disminución de la testosterona sérica y glutatión testicular, aumentos en el malondialdehído testicular, cambios degenerativos y apoptosis en células germinales, engrosamiento de la lámina basal tubular y aumentos en la expresión de la caspasa 3, y disminución en la expresión de PCNA. Los ARB (candesartán y losartán) revirtieron significativamente estos cambios con diferencias no significativas en el medio. El tratamiento con BRA (candesartán y losartán) revirtió significativamente la toxicidad testicular y cauda inducida por TAA en ratas. Esto podría ser potencialmente útil para el tratamiento temprano de pacientes con disfunción reproductiva inducida por tóxicos ocupacionales, especialmente si están usando BRA para otras comorbilidades.


Assuntos
Animais , Masculino , Ratos , Testículo/efeitos dos fármacos , Tioacetamida/toxicidade , Benzimidazóis/farmacologia , Losartan/farmacologia , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Testículo/patologia , Testosterona/análise , Tetrazóis/farmacologia , Imuno-Histoquímica , Ratos Sprague-Dawley , Antígeno Nuclear de Célula em Proliferação/metabolismo , Caspase 3/metabolismo , Glutationa/análise , Malondialdeído/análise
5.
BMC Plant Biol ; 19(1): 257, 2019 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-31200645

RESUMO

BACKGROUND: Proliferating cell nuclear antigen (PCNA), a conserved trimeric ring complex, is loaded onto replication fork through a hetero-pentameric AAA+ ATPase complex termed replication factor C (RFC) to maintain genome stability. Although architectures of PCNA-RFC complex in yeast have been revealed, the functions of PCNA and protein-protein interactions of PCNA-RFC complex in higher plants are not very clear. Here, essential regions mediating interactions between PCNA and RFC subunits in Arabidopsis and rice were investigated via yeast-two-hybrid method and bimolecular fluorescence complementation techniques. RESULTS: We observed that OsPCNA could interact with all OsRFC subunits, while protein-protein interactions only exist between Arabidopsis RFC2/3/4/5 and AtPCNA1/2. The truncated analyses indicated that the C-terminal of Arabidopsis RFC2/3/4/5 and rice RFC1/2 is essential for binding PCNA while the region of rice RFC3/4/5 mediating interaction with PCNA distributed both at the N- and C-terminal. On the other hand, we found that the C- and N-terminal of Arabidopsis and rice PCNA contribute equally to PCNA-PCNA interaction, and the interdomain connecting loop (IDCL) domain and C-terminal of PCNAs are indispensable for interacting RFC subunits. CONCLUSIONS: These results indicated that Arabidopsis and rice PCNAs are highly conserved in sequence, structure and pattern of interacting with other PCNA monomer. Nevertheless, there are also significant differences between the Arabidopsis and rice RFC subunits in binding PCNA. Taken together, our results could be helpful for revealing the biological functions of plant RFC-PCNA complex.


Assuntos
Arabidopsis/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteína de Replicação C/metabolismo , Arabidopsis/genética , Sequência Conservada , Oryza/genética , Proteínas de Plantas/genética , Antígeno Nuclear de Célula em Proliferação/genética
6.
J Microbiol Biotechnol ; 29(6): 863-876, 2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-31091863

RESUMO

Farm animals such as piglets are often affected by environmental stress, which can disturb the gut ecosystem. Antibiotics were commonly used to prevent diarrhea in weaned piglets, but this was banned by the European Union due to the development of antibiotic resistance. However, the use of probiotics instead of antibiotics may reduce the risk posed by pathogenic microorganisms and reduce the incidence of gastrointestinal diseases. Therefore, this study was conducted to investigate the effects of Lactobacillus casei Zhang on the mechanical barrier and immune function of early-weaned piglets infected using Escherichia coli K88 based on histomorphology and immunology. Fourteen-day-old weaned piglets were divided into a control group and experimental groups that were fed L. casei Zhang and infected with E. coli K88 with or without prefeeding and/or postfeeding of L. casei Zhang. The L. casei Zhang dose used was 107 CFU/g diet. Jejunum segments were obtained before histological, immunohistochemical, and western blot analyses were performed. In addition, the relative mRNA expression of toll receptors and cytokines was measured. Piglets fed L. casei Zhang showed significantly increased jejunum villus height, villus height-crypt depth ratio, muscle thickness, and expression of proliferating cell nuclear antigen and tight junction proteins ZO-1 and occludin. The use of L. casei Zhang effectively reduced intestinal inflammation after infection. We found that L. casei Zhang feeding prevented the jejunum damage induced by E. coli K88, suggesting that it may be a potential alternative to antibiotics for preventing diarrhea in early-weaned piglets.


Assuntos
Infecções por Escherichia coli/veterinária , Expressão Gênica/efeitos dos fármacos , Mucosa Intestinal/fisiologia , Jejuno/efeitos dos fármacos , Lactobacillus casei/fisiologia , Probióticos/administração & dosagem , Doenças dos Suínos/prevenção & controle , Animais , Citocinas/genética , Suplementos Nutricionais , Escherichia coli Enterotoxigênica/fisiologia , Infecções por Escherichia coli/patologia , Infecções por Escherichia coli/prevenção & controle , Fatores Imunológicos/genética , Fatores Imunológicos/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Jejuno/patologia , Ocludina/genética , Ocludina/metabolismo , Probióticos/farmacologia , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Suínos , Doenças dos Suínos/patologia , Receptores Toll-Like/genética , Desmame , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
7.
Reprod Biol Endocrinol ; 17(1): 36, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30982470

RESUMO

BACKGROUND: Endometriosis is an estrogen dependent, inflammatory disorder occurring in 5-10% of reproductive-aged women. Women with endometriosis have a lower body mass index (BMI) and decreased body fat compared to those without the disease, yet few studies have focused on the metabolic abnormalities in adipose tissue in women with endometriosis. Previously, we identified microRNAs that are differentially expressed in endometriosis and altered in the serum of women with the disease. Here we explore the effect of endometriosis on fat tissue and identified a role for endometriosis-related microRNAs in fat metabolism and a reduction in adipocyte stem cell number. METHODS: Primary adipocyte cells cultured from 20 patients with and without endometriosis were transfected with mimics and inhibitors of microRNAs 342-3p or Let 7b-5p to model the status of these microRNAs in endometriosis. RNA was extracted for gene expression analysis by qRT-PCR. PCNA expression was used as a marker of adipocyte proliferation. Endometriosis was induced experimentally in 9-week old female C57BL/6 mice and after 10 months fat tissue was harvested from both the subcutaneous (inguinal) and visceral (mesenteric) tissue. Adipose-derived mesenchymal stem cells in fat tissue were characterized in both endometriosis and non-endometriosis mice by FACS analysis. RESULTS: Gene expression analysis showed that endometriosis altered the expression of Cebpa, Cebpb, Ppar-γ, leptin, adiponectin, IL-6, and HSL, which are involved in driving brown adipocyte differentiation, appetite, insulin sensitivity and fat metabolism. Each gene was regulated by an alteration in microRNA expression known to occur in endometriosis. Analysis of the stem cell content of adipose tissue in a mouse model of endometriosis demonstrated a reduced number of adipocyte stem cells. CONCLUSIONS: We demonstrate that microRNAs Let-7b and miR-342-3p affected metabolic gene expression significantly in adipocytes of women with endometriosis. Similarly, there is a reduction in the adipose stem cell population in a mouse model of endometriosis. Taken together these data suggest that endometriosis alters BMI in part through an effect on adipocytes and fat metabolism.


Assuntos
Adipócitos/patologia , Endometriose/patologia , Adipócitos/metabolismo , Animais , Diferenciação Celular/genética , Proliferação de Células , Endometriose/genética , Endometriose/metabolismo , Feminino , Expressão Gênica , Humanos , Resistência à Insulina/genética , Metabolismo dos Lipídeos , Camundongos Endogâmicos C57BL , MicroRNAs/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo
8.
Toxicol Lett ; 310: 92-98, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30999038

RESUMO

Fullerenes have attracted attention since their discovery as structural units of complex carbon nanostructures capable of transporting drugs and macromolecules. As such artificial nanomaterials are applied in biology and medicine, they are routinely scrutinized for their effects on living organisms. The results of such studies range from direct destabilizing effects on DNA molecules to amelioration of the toxic effects of known genotoxic agents. We tested the effect of buckminsterfullerene (C60) on Drosophila melanogaster at DNA, tissue and organism levels. The water-soluble pristine C60 fullerene at the concentration of 20 µg/ml and 40 µg/ml leads to the activation of the mus209 gene in D. melanogaster larvae salivary glands, which can indicate higher levels of DNA damage. However, the absence of effects at the cell and organismal level could be explained by the activation of repair systems or by active elimination of damaged cells.


Assuntos
Drosophila melanogaster/efeitos dos fármacos , Fulerenos/toxicidade , Nanopartículas/toxicidade , Glândulas Salivares/efeitos dos fármacos , Animais , Animais Geneticamente Modificados , Dano ao DNA , Reparo do DNA/efeitos dos fármacos , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Feminino , Masculino , Testes de Mutagenicidade , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Medição de Risco , Glândulas Salivares/embriologia , Glândulas Salivares/metabolismo , Ativação Transcricional/efeitos dos fármacos
9.
Int J Oncol ; 54(6): 2019-2029, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30942439

RESUMO

Osteosarcoma (OS) is the most commonly diagnosed bone tumor in young people with poor prognosis. At present, the mechanisms underlying tumorigenesis in OS are not well understood. The methionine adnosyltransferase 2B (MAT2B) gene encodes the regulatory subunit of methionine adenosyltransferase (MAT). Recent studies demonstrated that it is highly expressed in a number of human malignancies; however, is undefined in OS. In the present study, MAT2B expression was investigated in tumor samples and cell lines. In vivo and in vitro, lentivirus­mediated small hairpin RNA was constructed to target the MAT2B gene and examine the role of MAT2B in OS proliferation. Microarray analysis was performed to examine the possible downstream molecular target of MAT2B in OS. MAT2B was markedly increased in OS specimens compared with the normal bone tissues, and it was additionally abundantly expressed in OS cell lines. Inhibition of MAT2B expression caused a marked decrease in proliferation and significant increase in apoptosis. In vivo, MAT2B silencing significantly inhibited OS cell growth. Microarray analysis suggested that epidermal growth factor receptor (EGFR) and proliferating cell nuclear antigen (PCNA) may function as downstream targets of MAT2B in OS, as confirmed by reverse transcription­quantitative polymerase chain reaction assays and western blotting. Collectively, these results suggested that MAT2B serves a critical role in the proliferation of OS by regulating EGFR and PCNA and that it may be a potential therapeutic target and prognostic factor of OS.


Assuntos
Neoplasias Ósseas/patologia , Metionina Adenosiltransferase/metabolismo , Osteossarcoma/patologia , Antígeno Nuclear de Célula em Proliferação/metabolismo , Animais , Apoptose/genética , Osso e Ossos/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Receptores ErbB/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Metionina Adenosiltransferase/genética , Camundongos , Camundongos Nus , Análise de Sequência com Séries de Oligonucleotídeos , Osteossarcoma/genética , RNA Interferente Pequeno/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
10.
Anim Reprod Sci ; 205: 44-51, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30981564

RESUMO

The bioflavonoid quercetin is a component of food with numerous biological effects, but its function in reproductive processes and mechanisms in various species remain unclear. The aim of this study was to examine the effect of quercetin on ovarian cells isolated from ovaries of two phytophagous mammalian species (i.e. pigs and cattle). There was analysis of the effect of quercetin (0, 1, 10, and 100 ng/mL) on cultured granulosa cells of pigs and cattle. Proliferation (PCNA) and apoptosis (bax) markers and release of progesterone (P4), testosterone (T), estradiol (E2), and IGF-I were quantified using quantitative immunocytochemistry, enzyme immunoassay, or radioimmunoassay. Treatments with quercetin reduced PCNA and bax accumulation and decreased P4 release from both granulosa cells of pigs and cattle. In cells of pigs, treatment with quercetin reduced T output, however, in cells of cattle quercetin increased T release. In cells of pigs, quercetin reduced IGF-I release. In cells of cattle, quercetin at smaller doses (1 or 10 ng/mL), promoted and at a large dose (100 ng/mL) reduced IGF-I secretions. There was no substantial E2 release from granulosa cells of pigs or cattle. These observations are the first to indicate there is a direct action of quercetin on basic ovarian cell functions (proliferation, apoptosis, and hormones release) which can be species-specific.


Assuntos
Bovinos , Células da Granulosa/efeitos dos fármacos , Quercetina/farmacologia , Suínos , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Estradiol/metabolismo , Feminino , Progesterona/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Especificidade da Espécie , Testosterona/metabolismo , Proteína X Associada a bcl-2
11.
Anim Reprod Sci ; 204: 140-151, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30948244

RESUMO

This study was conducted with the aim to understand the roles of apoptosis signal-regulating kinase (ASK1) and transcription factor tumor suppressor protein TP53, as well as the possible interrelationships, in the control of healthy ovarian cell functions. Rabbit ovarian granulosa cells were transfected with constructs encoding ASK1, TP53, or TP53 + ASK1 and cultured with or without insulin-like growth factor 1 (IGF1). The accumulation of ASK1, the cytoplasmic apoptosis regulators BAX and BCL2, and proliferating cell nuclear antigen (PCNA, a cell proliferation marker), as well as progesterone release, were evaluated by quantitative immunocytochemistry and radioimmunoassay. Results indicate both ASK1 and TP53 promoted the accumulation of BAX, but suppressed that of BCL2 and PCNA. Progesterone release was inhibited by ASK1 and promoted by TP53, while TP53 also stimulated ASK1 accumulation. Additionally, IGF1 stimulated PCNA and reduced progesterone release, but did not affect ASK1. Transfection with ASK1, TP53, or TP53 + ASK1 could modify IGF1 activity, however, there was no cumulative effect with co-transfection of TP53 and ASK1. This is the first results that indicate there is ASK1 suppression of healthy ovarian granulosa cell functions, including promoting apoptosis, inhibiting proliferation, and alter progesterone release. There was also TP53 actions in rabbit ovarian granulosa cells, where it stimulated ASK1, apoptosis, and progesterone release, thus suppressing proliferation and responses to IGF1. The similarity of ASK1 and TP53 effects on apoptosis and proliferation, lack of cumulative action of these molecules, and capacity of TP53 to promote ASK1 accumulation suggest that TP53 can suppress some ovarian granulosa cell functions through ASK1 stimulation.


Assuntos
Células da Granulosa/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Coelhos , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/fisiologia , Biomarcadores , Sobrevivência Celular , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Fluorescência Verde , Transtornos do Crescimento , Perda Auditiva Neurossensorial , Fator de Crescimento Insulin-Like I/deficiência , Fator de Crescimento Insulin-Like I/farmacologia , MAP Quinase Quinase Quinase 5/genética , Progesterona , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
12.
Cell Physiol Biochem ; 52(3): 421-434, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30845381

RESUMO

BACKGROUND/AIMS: The aim of this study was to evaluate the potential and significant applications of Sertoli cells (SCs) transplantation, and to explore the effect of transplantation on spermatogenesis process, in azospermic mice. METHODS: In this study, we utilized 18 adult mice (28‒30 g), divided into four experimental groups: (1) control, (2) vehicle (DMSO 2%) (10 µl) (3) busulfan and (4) busulfan+ SCs (1×104 cells/µL). SCs were isolated from the testis of 4-week-old mouse and after using anesthetics, 10 µl of SCs suspension (1×104 cells/µL) was injected over 3-5 min, into each testis and subsequently, sperm samples were collected from the tail of the epididymis. Afterward, the animals were euthanized and testis samples were taken for histopathology experiments, and RNA extraction, in order to examine the expression of c-kit, STRA8 and PCNA genes. RESULTS: Our data showed that SCs transplantation could notably increase the total sperm count and the number of testicular cells, such as spermatogonia, primary spermatocyte, round spermatid, SCs and Leydig cells, compared to the control, DMSO and busulfan groups. Furthermore, the result showed that the expression of c-kit and STRA8 were significantly decreased in busulfan and busulfan/SCs groups, at 8 weeks after the last injection (p<0.001), but no significant decrease was found for PCNA, compared to the control and DMSO groups (P<0.05). CONCLUSION: These findings suggest that SCs transplantation may be beneficial as a practical approach for therapeutic strategies in reproductive and regenerative medicine. We further highlighted the essential applications that might provide a mechanism for correcting fertility in males, suffering from cell deformity.


Assuntos
Células de Sertoli/transplante , Espermatogênese , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Bussulfano/farmacologia , Epididimo/citologia , Epididimo/metabolismo , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/metabolismo , Masculino , Camundongos , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/metabolismo , Medicina Regenerativa , Células de Sertoli/citologia , Motilidade Espermática , Espermátides/citologia , Espermátides/metabolismo , Espermatogênese/efeitos dos fármacos , Espermatogônias/citologia , Espermatogônias/fisiologia , Testículo/metabolismo , Testículo/patologia
13.
Gene ; 700: 31-37, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-30898712

RESUMO

We investigated changes in expression of the CIP/KIP family-related genes and the cycle-dependent factors Pcna, Cdk4 and Cdk2 during the growth and development of mice, Drosophila and silkworms. When the organism was in a period of rapid development, the related genes of the CIP/KIP family had low expression level and the cell cycle-dependent genes were highly expressed. In mammals, the CIP/KIP family includes three genes, p21, p27/Dacapo and p57. However, only one gene, P27/Dacapo, exists in the CIP/KIP family in silkworm and the orthologous gene in the silkworm is named Bmdacapo. Down-regulation of Bmdacapo in silkworm embryos caused overdevelopment of the embryos and indicated that Bmdacapo can inhibit silkworm growth and development. Up-regulation of Bmdacapo in silkworm cells inhibited cell proliferation, whereas down-regulation of Bmdacapo promoted cell proliferation. In order to explore the mechanism of Bmdacapo regulated silkworm development and cell proliferation, the effect of Bmdacapo on cell cycle changes was examined. The results demonstrate that Bmdacapo was able to induce G1/S phase arrest in the cell cycle. In silkworm cells, Bmdacapo inhibits the expression of Pcna, CDK4 and CDK2, which affects the cell cycle and ultimately inhibits cell proliferation. This regulatory mechanism is particularly different from mammals.


Assuntos
Bombyx/embriologia , Inibidor de Quinase Dependente de Ciclina p27/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Drosophila melanogaster/embriologia , Animais , Bombyx/citologia , Bombyx/metabolismo , Ciclo Celular , Proliferação de Células , Células Cultivadas , Clonagem Molecular , Quinase 2 Dependente de Ciclina/genética , Quinase 2 Dependente de Ciclina/metabolismo , Quinase 4 Dependente de Ciclina/genética , Quinase 4 Dependente de Ciclina/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Desenvolvimento Embrionário , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Camundongos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo
14.
Theriogenology ; 131: 1-8, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30921633

RESUMO

Continental waters salinisation is a global threat that has grown because of climate change and human activities, but little is known about how and what biological tracts are affected. The aim of this study was to investigate the influence of different water salinities on the expression of HSP70, PCNA and caspase-3 during spermatogenesis of Nile tilapia. Adult males were submitted to four salinity treatments: (S0) fresh water, (S7) 7 g L-1, (S14) 14 g L-1, and (S21) 21 g L-1 for 1, 4, and 9 days. All specimens were in spermatogenic activity and the highest values of the gonadosomatic index (GSI) occurred in the S0 and S7. In the morphometric analysis, spermatocytes were the most frequent germ cell detected in all treatments (>50%) and spermatids achieved about 20% of the testicular proportion, with few variations among treatments. Spermatozoa were significantly reduced only in S14 compared to S7. Leydig cells were significantly increased in S14 when compared to S7 but plasma concentrations of 11-KT showed no significant difference among treatments. ELISA assay showed higher testicular expression of HSP70 at 1 day in all groups, followed by a significant decrease at days 4 and 9 in S14 and S21. The expression of PCNA was significantly lower while the activity of caspase-3 was higher in S14 and S21 when compared to S0 and S7. These results indicate that higher salinities in S14 and S21 interfere with the relationship between testicular HSP70, PCNA, and caspase-3, but with few effects over spermatogenesis dynamics of Nile tilapia.


Assuntos
Salinidade , Espermatogênese/fisiologia , Testículo/efeitos dos fármacos , Tilápia/fisiologia , Animais , Caspase 3/metabolismo , Mudança Climática , Ensaio de Imunoadsorção Enzimática , Proteínas de Peixes/metabolismo , Proteínas de Choque Térmico HSP72/metabolismo , Masculino , Antígeno Nuclear de Célula em Proliferação/metabolismo , Testículo/metabolismo , Testículo/fisiologia , Testosterona/análogos & derivados , Testosterona/sangue , Tilápia/metabolismo
15.
Int J Biol Macromol ; 130: 307-314, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30825564

RESUMO

A comparison of the anti-tumor activity of CMPS-II and CBPS-II polysaccharides, respectively is obtained from the fermented mycelium and cultivated fruiting bodies of Cordyceps militaris. This in vitro anti-tumor activity is investigated using an MTT assay, immunofluorescence staining, a Western Blot assay, a qRT-PCR assay, and Annexin V-FITC/PI double staining. The experimental results indicate that the inhibition rate of CMPS-II on H1299 tumor cells is higher than that of CBPS-II. With a concentration of 500 µâ€¯g/mL, the inhibition rate of CMPS-II and CBPS-II were 54.55% and 34.80%, respectively. Both CMPS-II and CBPS-II can increase the protein and mRNA expression level of cell apoptosis factors Caspase-3, Caspase-9, and p53, while reducing the protein and mRNA expression levels of proliferating cell nuclear antigen (PCNA), to induce tumor cells apoptosis. The induction effect of CMPS-II was stronger than CBPS-II. These results suggest that CMPS-II is superior to CBPS-II regarding the inhibition of H1299 lung cancer cells. Furthermore, CMPS-II is a potentially useful substitution for CBPS-II in the treatment of lung cancer and provides new insights into the mechanism of its anti-tumor activity.


Assuntos
Antineoplásicos/farmacologia , Cordyceps/metabolismo , Fermentação , Carpóforos/metabolismo , Polissacarídeos Fúngicos/farmacologia , Micélio/metabolismo , Antineoplásicos/metabolismo , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Polissacarídeos Fúngicos/biossíntese , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
16.
Oxid Med Cell Longev ; 2019: 2471312, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30906501

RESUMO

Accumulation of oxidative insults on molecular and supramolecular levels could compromise renewal potency and architecture in the aging skin. To examine and compare morphological and ultrastructural changes with redox alterations during chronological skin aging, activities of antioxidant defense (AD) enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), thioredoxin reductase (TR), and methionine sulfoxide reductase A (MsrA), and the markers of oxidative damage of biomolecules-4-hydroxynonenal (HNE) and 8-oxoguanine (8-oxoG)-were examined in the rat skin during life (from 3 days to 21 months). As compared to adult 3-month-old skin, higher activities of CAT, GSH-Px, and GR and a decline in expression of MsrA are found in 21-month-old skin. These changes correspond to degenerative changes at structural and ultrastructural levels in epidermal and dermal compartments, low proliferation capacity, and higher levels of HNE-modified protein aldehydes (particularly in basal lamina) and 8-oxoG positivity in nuclei and mitochondria in the sebaceous glands and root sheath. In 3-day-old skin, higher activities of AD enzymes (SOD, CAT, GR, and TR) and MsrA expression correspond to intensive postnatal development and proliferation. In contrast to 21-month-old skin, a high level of HNE in young skin is not accompanied by 8-oxoG positivity or any morphological disturbances. Observed results indicate that increased activity of AD enzymes in elderly rat skin represents the compensatory response to accumulated oxidative damage of DNA and proteins, accompanied by attenuated repair and proliferative capacity, but in young rats the redox changes are necessary and inherent with processes which occur during postnatal skin development. Мorphological and ultrastructurаl changes are in line with the redox profile in the skin of young and old rats.


Assuntos
Envelhecimento/metabolismo , Pele/metabolismo , Pele/patologia , Aldeídos/metabolismo , Animais , Catalase/metabolismo , Proliferação de Células , Colágeno/ultraestrutura , Dano ao DNA , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Fibroblastos/ultraestrutura , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Masculino , Oxirredução , Oxirredutases/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos Wistar , Pele/enzimologia , Pele/ultraestrutura , Superóxido Dismutase/metabolismo , Tiorredoxina Dissulfeto Redutase/metabolismo , Fatores de Tempo
17.
Can J Gastroenterol Hepatol ; 2019: 9015453, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30881947

RESUMO

Background: Hepatocellular carcinoma (HCC) is one of the most common malignant cancers with a poor prognosis. Several commonly investigated immunohistochemical markers in resected HCC have potential prognostic value, but the prognostic utility of p53 expression in HCC has remained elusive. Aim: To evaluate the prognostic value of p53 and p53 phosphorylation at serine 15 (p53 Ser15-P) in patients with HCC. Methods: Surgically resected tumors from 199 HCC patients were analyzed for p21, p53, p53 Ser15-P, and proliferating cell nuclear antigen (PCNA) expression using immunohistochemistry. Results: Stratifying by the expression of p53 Ser15-P (P = 0.016), but not by p53 (P = 0.301), revealed significantly different survival outcomes in patients with HCC. Moreover, our analysis demonstrated that patients who were PCNA-positive and p53 Ser15-P-negative had significantly worse survival outcomes (P = 0.001) than patients who were PCNA-positive and p53 Ser15-P-positive. Conclusions: P53 Ser15-P is associated with poor outcomes in patients with HCC, and this prognostic marker is useful for predicting the survival of patients with PCNA-positive HCC.


Assuntos
Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/cirurgia , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/cirurgia , Masculino , Pessoa de Meia-Idade , Fosforilação , Prognóstico , Taxa de Sobrevida
18.
Plant Sci ; 280: 297-304, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30824007

RESUMO

The Proliferating Cell Nuclear Antigen, PCNA, has roles in both G1 and S phases of the cell cycle. Here we show that maize PCNA can be found in cells in structures of a trimer or a dimer of trimer, in complexes of high molecular mass that change in size as germination proceeds, co-eluting with cell cycle proteins as CycD3;1 and CDKs (A/B1;1). Using different methodological strategies, we show that PCNA actually interacts with CycD3;1, CDKA, CDKB1;1, KRP1;1 and KRP4;1, all of which contain PIP or PIP-like motifs. Anti-PCNA immunoprecipitates show kinase activity that is inhibited by KRP1;1 and KRP4;2, indicating the formation of quaternary complexes PCNA-CycD/CDKs-KRPs in which PCNA would act as a platform. This inhibitory effect seems to be differential during the germination process, more pronounced as germination advances, suggesting a complex regulatory mechanism in which PCNA could bind different sets of cyclins/CDKs, some more susceptible to inhibition by KRPs than others.


Assuntos
Quinases Ciclina-Dependentes/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Zea mays/genética , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Quinases Ciclina-Dependentes/genética , Ciclinas/genética , Ciclinas/metabolismo , Germinação , Fosforilação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Antígeno Nuclear de Célula em Proliferação/genética , Zea mays/enzimologia , Zea mays/fisiologia
19.
Niger J Clin Pract ; 22(2): 194-200, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30729942

RESUMO

Background: Colorectal cancers are third most common cancer in both genders. They are associated with genetic and environmental factors. Staging is important in the prognosis. Carcinoembryonic Antigen (CEA) provides preliminary information and there is a correlation between Proliferation Index (PI) and prognostic variables. Our aim is to investigate the relationship between DNA repair capacity and clinico-pathologic factors. Patients and Methods: The blood samples taken from cancer patients were irradiated. DNA repair capacity by comet technique was calculated. The CEA values were recorded. Pathology reports were collected and PI values were calculated. s. Results: Total of 30 patients; male (n: 14) and female (n: 16) with a median age of 66.37 ± 10.32 were included. Mean CEA value was 42.85 (1.46 - 422.30 µgr/ml) µgr/ml. Mean % DNA repair capacity was 44.49 ± 5.24. In the pathology; 21 (70%) were T3 tumors; 18 (60%) had lymphatic and 12 (40%) had vascular 2 invasion. Perineural invasion was present in 8 (26.7%). According to the proliferation index (PI); 16 (53.3%) were in high percentile (PI > 66%) group. Conclusions: There was a significant correlation between; perineural invasion and tumor grade (P = 0.043); lymphatic and perineural invasion (P = 0.006); lymphatic invasion and vascular invasion (P = 0.034) and the DNA repair capacity with the lymphatic invasion (P = 0.026). There was also a statistically significant (P = 0.044) relationship between PI and lymphatic invasion. As a result in colorectal cancer patients DNA repair capacity can be used as a biomarker in the staging and also in the prediction of the tumor behavior.


Assuntos
Antígeno Carcinoembrionário/sangue , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Reparo do DNA , DNA de Neoplasias/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Idoso , Núcleo Celular/química , Neoplasias Colorretais/sangue , Neoplasias Colorretais/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Prognóstico , Estudos Prospectivos
20.
J Int Med Res ; 47(1): 427-437, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30791830

RESUMO

OBJECTIVE: This study was performed to evaluate the impact and underlying mechanisms of hypothermic machine perfusion (HMP) on half-size liver graft regeneration. METHODS: Forty rats were randomly assigned to five groups: two in vitro groups (static cold storage [SCS] and HMP) and three in vivo groups (orthotopic liver transplantation, SCS, and HMP). Perfusates and plasma samples were collected for analysis of hepatic enzymes. Liver tissue was obtained for evaluation of histology, immunohistochemistry (Ki67 and proliferating cell nuclear antigen [PCNA]), and the regeneration rate. Cell cycle genes were analyzed by quantitative real-time polymerase chain reaction, and cyclin D1 and cyclin E1 were semiquantified by western blot. RESULTS: HMP improved histopathological outcomes and decreased hepatic enzyme release. The expression of Ki67 and PCNA demonstrated a greater proliferation activity in the HMP than SCS group, and the expression of almost all cell cycle genes was elevated following HMP. Western blot results showed higher protein levels of cyclin D1 and cyclin E1 in the HMP than SCS group. CONCLUSIONS: Our findings suggest for the first time that half-size liver graft protection by HMP involves recovery of graft regeneration.


Assuntos
Transplante de Fígado/métodos , Fígado/cirurgia , Perfusão/métodos , Regeneração/fisiologia , Animais , Temperatura Baixa , Ciclina D1/genética , Ciclina D1/metabolismo , Ciclinas/genética , Ciclinas/metabolismo , Expressão Gênica , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Fígado/metabolismo , Testes de Função Hepática , Masculino , Soluções para Preservação de Órgãos/farmacologia , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos , Ratos Sprague-Dawley , Transplantes/efeitos dos fármacos , Transplantes/metabolismo
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