ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of total flavanones of Sedum sarmentosum (SSTF) on the apoptosis of rat hepatic stellate cells (HSC-T6) and its mechanism.</p><p><b>METHOD</b>Different concentrations of SSTF and HSC-T6 cells were co-cultured for different period of time. The MTT assay was used to detect the inhibitory effect of SSTF on the proliferation of HSC-T6 cells. The flow cytometry Annexin-V/PI double staining method was adopted to detect SSTF's effect on HSC-T6 cell apoptosis. Western blotting and Real-time PCR methods were applied to observe the effect on the protein and mRNA expressions of apoptosis-related cytokines Bcl-2, Bax and Caspase-3.</p><p><b>RESULT</b>SSTF significantly inhibited HSC-T6 cell proliferation and induced cell apoptosis in a dose and time dependent manner. According to Western blotting result, SSTF promoted apoptosis by inhibiting Bcl-2, Bax and promoting the protein expression of Caspase-3; according to a further Real-time PCR study, Bcl-2 mRNA levels can inhibit Bcl-2 and promote Bax and Caspase-3 expressions.</p><p><b>CONCLUSION</b>SSTF has the effect of promoting the apoptosis of HSC-T6 mainly by inhibiting Bcl-2 and promoting protein and mRNA expressions of Bax and caspase-3.</p>
Subject(s)
Animals , Rats , Apoptosis , Caspase 3 , Genetics , Metabolism , Cell Line , Cell Proliferation , Drugs, Chinese Herbal , Pharmacology , Flavanones , Pharmacology , Hepatic Stellate Cells , Cell Biology , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Sedum , ChemistryABSTRACT
The growth characteristics and ginsenosides isolation of the suspension-cultured crown gall of Panax quinquefolium were studied. The result showed that the maximum biomass of cultures was 18.6 g/L (dry weight) and the content of ginsenosides reached its maximum level of 620.4 mg/L on the 27th day. The utilization rates of sugar, phosphorus, nitrogen in NH4+ and nitrogen in NO3- were 91.8%, 100%, 81% and 97%, respectively. Four compounds were isolated from the suspension-cultured crown gall and their structures were elucidated as pseudoginsenoside F11 (I), ginsenoside Rd (II), ginsenoside Rb1 (llI) and ginsenoside Rb3 (IV).