Objective:To investigate apoptotic effects of berberine,a significant alkaloids component existing in Rhizoma coptidis,and its possible acting mechanism in insulinoma cells.Methods:Different concentrations of berberin...Objective:To investigate apoptotic effects of berberine,a significant alkaloids component existing in Rhizoma coptidis,and its possible acting mechanism in insulinoma cells.Methods:Different concentrations of berberine were used to treat mouse insulinoma(MIN6)cells for various period of time.The viability and apoptosis of the cells were analyzed using methylthiazolyldiphenvl-tetrazolium bromide assay,flow cytometry and enzyme-linked immuno sorbent assay.Changes in the relating pro-and anti-apoptosis proteins were detected by western-blotting.Results:The half-maximal inhibitory concentration(IC50)of berberine was 5.7μmol/L on MIN6 cells viability for 16 h.Berberine caused a 20%reduction(P<0.05)in cell number after only 4-h incubation;which reached 50%after 24 h(P<0.01).Berberine treatment for 16 h significantly increased the level of DNA fragmentation.The flow cytometry showed the apoptotic rate increased 2.9-and 4.6-fold after treating with berberine(5μmol/L)for 8 and 16 h,while 3-and 8.7-fold after 10μmol/L treatment for 8 and 16 h(P<0.01).Berberine treatment dramatically elevated the expression ratio of Bax to Bcl-2.Meanwhile,berberine notably increased the apoptosis-inducing factors and cytochrome C transforming from the mitochondria to the cytoplasm.Apoptotic protease-activating factor 1(Apaf-1)was subsequently activated after cytochrome C release.Furthermore,caspase-3 and poly adenosine diphosphate-ribose polymerase were also activated to trigger apoptosis cascade.Conclusion:High concentration(5 and 10μmol/L)of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor(AIF)pathway.展开更多
基金Supported by the National Natural Science Foundation of China(No.NSFC81274041 and No.NSFC81273995)the International Cooperation Projects of Ministry of Education(No.2011DFA30920)the Key Drug Development Program of Ministry of Science an Technology(No.20122X09103201-005)
文摘Objective:To investigate apoptotic effects of berberine,a significant alkaloids component existing in Rhizoma coptidis,and its possible acting mechanism in insulinoma cells.Methods:Different concentrations of berberine were used to treat mouse insulinoma(MIN6)cells for various period of time.The viability and apoptosis of the cells were analyzed using methylthiazolyldiphenvl-tetrazolium bromide assay,flow cytometry and enzyme-linked immuno sorbent assay.Changes in the relating pro-and anti-apoptosis proteins were detected by western-blotting.Results:The half-maximal inhibitory concentration(IC50)of berberine was 5.7μmol/L on MIN6 cells viability for 16 h.Berberine caused a 20%reduction(P<0.05)in cell number after only 4-h incubation;which reached 50%after 24 h(P<0.01).Berberine treatment for 16 h significantly increased the level of DNA fragmentation.The flow cytometry showed the apoptotic rate increased 2.9-and 4.6-fold after treating with berberine(5μmol/L)for 8 and 16 h,while 3-and 8.7-fold after 10μmol/L treatment for 8 and 16 h(P<0.01).Berberine treatment dramatically elevated the expression ratio of Bax to Bcl-2.Meanwhile,berberine notably increased the apoptosis-inducing factors and cytochrome C transforming from the mitochondria to the cytoplasm.Apoptotic protease-activating factor 1(Apaf-1)was subsequently activated after cytochrome C release.Furthermore,caspase-3 and poly adenosine diphosphate-ribose polymerase were also activated to trigger apoptosis cascade.Conclusion:High concentration(5 and 10μmol/L)of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor(AIF)pathway.
