[Objective] To investigate the protective effects of Aplysin on ethanol-induced oxidative damage in rat primary hepatocytes.[Methods]Rat primary hepatocytes were obtained via the portal vein collagenase IV in situ per...[Objective] To investigate the protective effects of Aplysin on ethanol-induced oxidative damage in rat primary hepatocytes.[Methods]Rat primary hepatocytes were obtained via the portal vein collagenase IV in situ perfusion technique followed by a Percoll density gradient centrifuge; MTT test was used to determine the optimum dose of Aplysin and ethanol,and detect the cell vitality in primary hepatocytes; supernatants of primary hepatocytes were harvested to measure AST and LDH level,and the SOD,GSH-PX activities and MDA content in primary hepatocytes were observed; flow cytometry was used to detect the cell apoptosis rate; DNA damage in primary hepatocytes were detected by single-cell gel electrophoresis assay; The level of mitochondrial membrane potential in primary hepatocytes was tested by fluorogenic probe JC-1; the CYP2E1 activity in primary hepatocytes were detected by colorimetry; the proteins of CYP2E1 were detected by Western blotting. [Results]300 mmol/L dose of ethanol and 30 mg/L dose of Aplysin were the optimal dosages and were used in the subsequent experiments. Hepatocyte vitality was significantly increased in the Aplysin group relative to that in the ethanol group,and Aplysin inhibited the release of AST and LDH( P < 0. 05). For the Aplysin treatment group,the activities of hepatocyte SOD and GSH were significantly increased and MDA was markedly lowered as compared with those in the ethanol group( P < 0. 05). And Aplysin can alleviate hepatocyte apoptosis significantly,and hepatocyte DNA damage rate of II-III level and IV level were significantly lowered in the Aplysin treatment group as compared with those in the ethanol group,and Aplysin has evidently improved on alcohol induced mitochondria damage of hepatocyte. Primary hepatocytes activities and protein expression of CYP2E1 were markedly lowered in the Aplysin treatment group as compared with those in the ethanol group( P < 0. 05). [Conclusion] Aplysin has protective effects on liver oxidative damage induced by alcohol of primary cultured rat hepatocytes by blocking CYP2E1 activation,relieving oxidative stress,and sharpening the oxidation resistance ability.展开更多
基金Supported by Natural Science Foundation of China(81573137)
文摘[Objective] To investigate the protective effects of Aplysin on ethanol-induced oxidative damage in rat primary hepatocytes.[Methods]Rat primary hepatocytes were obtained via the portal vein collagenase IV in situ perfusion technique followed by a Percoll density gradient centrifuge; MTT test was used to determine the optimum dose of Aplysin and ethanol,and detect the cell vitality in primary hepatocytes; supernatants of primary hepatocytes were harvested to measure AST and LDH level,and the SOD,GSH-PX activities and MDA content in primary hepatocytes were observed; flow cytometry was used to detect the cell apoptosis rate; DNA damage in primary hepatocytes were detected by single-cell gel electrophoresis assay; The level of mitochondrial membrane potential in primary hepatocytes was tested by fluorogenic probe JC-1; the CYP2E1 activity in primary hepatocytes were detected by colorimetry; the proteins of CYP2E1 were detected by Western blotting. [Results]300 mmol/L dose of ethanol and 30 mg/L dose of Aplysin were the optimal dosages and were used in the subsequent experiments. Hepatocyte vitality was significantly increased in the Aplysin group relative to that in the ethanol group,and Aplysin inhibited the release of AST and LDH( P < 0. 05). For the Aplysin treatment group,the activities of hepatocyte SOD and GSH were significantly increased and MDA was markedly lowered as compared with those in the ethanol group( P < 0. 05). And Aplysin can alleviate hepatocyte apoptosis significantly,and hepatocyte DNA damage rate of II-III level and IV level were significantly lowered in the Aplysin treatment group as compared with those in the ethanol group,and Aplysin has evidently improved on alcohol induced mitochondria damage of hepatocyte. Primary hepatocytes activities and protein expression of CYP2E1 were markedly lowered in the Aplysin treatment group as compared with those in the ethanol group( P < 0. 05). [Conclusion] Aplysin has protective effects on liver oxidative damage induced by alcohol of primary cultured rat hepatocytes by blocking CYP2E1 activation,relieving oxidative stress,and sharpening the oxidation resistance ability.
