AIM: To investigate the co-incidence of apoptosis, autophagy, and unfolded protein response(UPR) in hepatitis B(HBV) and C(HCV) infected hepatocytes.METHODS: We performed immunofluorescence confocal microscopy on 10 l...AIM: To investigate the co-incidence of apoptosis, autophagy, and unfolded protein response(UPR) in hepatitis B(HBV) and C(HCV) infected hepatocytes.METHODS: We performed immunofluorescence confocal microscopy on 10 liver biopsies from HBV and HCV patients and tissue microarrays of HBV positive liver samples. We used specific antibodies for LC3β, cleaved caspase-3, BIP(GRP78), and XBP1 to detect autophagy, apoptosis and UPR, respectively. AntiHCV NS3 and anti-HBs antibodies were also used to confirm infection. We performed triple blind counting of events to determine the co-incidence of autophagy(LC3β punctuate), apoptosis(cleaved caspase-3), and unfolded protein response(GRP78) with HBV and HCV infection in hepatocytes. All statistical analyses were performed using SPSS software for Windows(Version 16 SPSS Inc, Chicago, IL, United States). P-values < 0.05 were considered statistically significant. Statistical analyses were performed with Mann-Whitney test to compare incidence rates for autophagy, apoptosis, and UPR in HBV- and HCV-infected cells and adjacent noninfected cells.RESULTS: Our results showed that infection of hepatocytes with either HBV and HCV induces significant increase(P < 0.001) in apoptosis(cleavage of caspase-3), autophagy(LC3β punctate), and UPR(increase in GRP78 expression) in the HCV- and HBVinfected cells, as compared to non-infected cells of the same biopsy sections. Our tissue microarray immunohistochemical expression analysis of LC3β in HBV^(Neg) and HBV^(Pos) revealed that majority of HBVinfected hepatocytes display strong positive stainingfor LC3β. Interestingly, although XBP splicing in HBVinfected cells was significantly higher(P < 0.05), our analyses show a slight increase of XBP splicing was in HCV-infected cells(P > 0.05). Furthermore, our evaluation of patients with HBV and HCV infection based on stage and grade of the liver diseases revealed no correlation between these pathological findings and induction of apoptosis, autophagy, and UPR.CONCLUSION: The results of this study indicate that HCV and HBV infection activates apoptosis, autophagy and UPR, but slightly differently by each virus. Further studies are warranted to elucidate the interconnections between these pathways in relation to pathology of HCV and HBV in the liver tissue.展开更多
AIM:The most important side effect of methotrexate(MTX)is mucositis.The purpose of this study was to evaluate the effect of turmeric extract on intestinal damage and oxidative stress in rats receiving methotrexate.MET...AIM:The most important side effect of methotrexate(MTX)is mucositis.The purpose of this study was to evaluate the effect of turmeric extract on intestinal damage and oxidative stress in rats receiving methotrexate.METHODS:Experiments were performed on male Wistar albino rats divided into six groups.First group received normal saline orally,the second group received turmeric extract(100 mg·kg-1)orally for 30 days,the third group received turmeric extract(200 mg·kg)orally for 30 days,the fourth group received a single dose of methotrexate(20 mg·kg)i.p.at day 30,the fifth group received turmeric extract(100 mg·kg)orally for 30 days and a single dose of methotrexate(20 mg·kg)i.p.at day 30,and the sixth group received turmeric extract(200 mg·kg)orally for 30days and single dose of methotrexate(20 mg·kg)i.p.at day 30.Four days after methotrexate injection,animals were anesthetized,blood samples were taken to determine total antioxidant status(TAS)and jejunum samples were taken for glutathione peroxidase(GPx),superoxidase dismutase(SOD),catalase(CAT),aldehyde malondialdehyde(MDA),and histopathological assessment.RESULTS: Microscopic evaluation from intestinal tissues of the MTX treated group,showed severe villus shortening and blunting,inflammatory cell infiltration and hemorrhage in lamina propria,along with epithlial cell necrosis.Levels of SOD,GSH-Px and CAT decreased in the MTX received group,but increased significantly(P<0.05)in the turmeric+MTX groups.MTX increased lipid peroxidation,however,turmeric decreased peroxidation significantly(P<0.05).CONCLUSION:These results suggest that turmeric extract may protect the small intestine of rats from methotrexate-induced damage.Turmeric effects could result from its antioxidant properties.展开更多
基金Supported by University of Manitoba Start-up funds and an award from the Manitoba Medical Service Foundation to Ghavami SUniversity of Manitoba Start-up Funds to Alizadeh J
文摘AIM: To investigate the co-incidence of apoptosis, autophagy, and unfolded protein response(UPR) in hepatitis B(HBV) and C(HCV) infected hepatocytes.METHODS: We performed immunofluorescence confocal microscopy on 10 liver biopsies from HBV and HCV patients and tissue microarrays of HBV positive liver samples. We used specific antibodies for LC3β, cleaved caspase-3, BIP(GRP78), and XBP1 to detect autophagy, apoptosis and UPR, respectively. AntiHCV NS3 and anti-HBs antibodies were also used to confirm infection. We performed triple blind counting of events to determine the co-incidence of autophagy(LC3β punctuate), apoptosis(cleaved caspase-3), and unfolded protein response(GRP78) with HBV and HCV infection in hepatocytes. All statistical analyses were performed using SPSS software for Windows(Version 16 SPSS Inc, Chicago, IL, United States). P-values < 0.05 were considered statistically significant. Statistical analyses were performed with Mann-Whitney test to compare incidence rates for autophagy, apoptosis, and UPR in HBV- and HCV-infected cells and adjacent noninfected cells.RESULTS: Our results showed that infection of hepatocytes with either HBV and HCV induces significant increase(P < 0.001) in apoptosis(cleavage of caspase-3), autophagy(LC3β punctate), and UPR(increase in GRP78 expression) in the HCV- and HBVinfected cells, as compared to non-infected cells of the same biopsy sections. Our tissue microarray immunohistochemical expression analysis of LC3β in HBV^(Neg) and HBV^(Pos) revealed that majority of HBVinfected hepatocytes display strong positive stainingfor LC3β. Interestingly, although XBP splicing in HBVinfected cells was significantly higher(P < 0.05), our analyses show a slight increase of XBP splicing was in HCV-infected cells(P > 0.05). Furthermore, our evaluation of patients with HBV and HCV infection based on stage and grade of the liver diseases revealed no correlation between these pathological findings and induction of apoptosis, autophagy, and UPR.CONCLUSION: The results of this study indicate that HCV and HBV infection activates apoptosis, autophagy and UPR, but slightly differently by each virus. Further studies are warranted to elucidate the interconnections between these pathways in relation to pathology of HCV and HBV in the liver tissue.
基金supported by the Research Fund of Ardabil University of Medical Sciences
文摘AIM:The most important side effect of methotrexate(MTX)is mucositis.The purpose of this study was to evaluate the effect of turmeric extract on intestinal damage and oxidative stress in rats receiving methotrexate.METHODS:Experiments were performed on male Wistar albino rats divided into six groups.First group received normal saline orally,the second group received turmeric extract(100 mg·kg-1)orally for 30 days,the third group received turmeric extract(200 mg·kg)orally for 30 days,the fourth group received a single dose of methotrexate(20 mg·kg)i.p.at day 30,the fifth group received turmeric extract(100 mg·kg)orally for 30 days and a single dose of methotrexate(20 mg·kg)i.p.at day 30,and the sixth group received turmeric extract(200 mg·kg)orally for 30days and single dose of methotrexate(20 mg·kg)i.p.at day 30.Four days after methotrexate injection,animals were anesthetized,blood samples were taken to determine total antioxidant status(TAS)and jejunum samples were taken for glutathione peroxidase(GPx),superoxidase dismutase(SOD),catalase(CAT),aldehyde malondialdehyde(MDA),and histopathological assessment.RESULTS: Microscopic evaluation from intestinal tissues of the MTX treated group,showed severe villus shortening and blunting,inflammatory cell infiltration and hemorrhage in lamina propria,along with epithlial cell necrosis.Levels of SOD,GSH-Px and CAT decreased in the MTX received group,but increased significantly(P<0.05)in the turmeric+MTX groups.MTX increased lipid peroxidation,however,turmeric decreased peroxidation significantly(P<0.05).CONCLUSION:These results suggest that turmeric extract may protect the small intestine of rats from methotrexate-induced damage.Turmeric effects could result from its antioxidant properties.