垂盆草总黄酮影响肝星状细胞凋亡的作用机制研究  

Study on effect of total flavanones of Sedum sarmentosum on apoptosis of hepatic stellate cells and its mechanism

作　　者：林远灿[1] 骆海莺[2] 金乾兴[1] 

机构地区：[1]浙江中医药大学附属第一医院,浙江杭州310006 [2]南京军区杭州疗养院,浙江杭州310007

出　　处：《中国中药杂志》2015年第16期3273-3277,共5页China Journal of Chinese Materia Medica

基　　金：浙江省自然科学基金项目(LQ15H290001);浙江中医药大学科研基金项目(2014ZY08)

摘　　要：目的:研究垂盆草总黄酮(SSTF)对大鼠肝星状细胞(HSC-T6)凋亡的影响及其作用机制。方法:将不同质量浓度的SSTF与HSC-T6细胞共培养不同时间后,MTT法检测SSTF对HSC-T6细胞的增殖抑制作用;流式细胞仪Annexin-V/PI双染方法检测SSTF对HSC-T6细胞凋亡的影响;应用Western blotting和Real-time PCR方法观察药物对凋亡相关细胞因子Bcl-2,Bax和Caspase-3蛋白及mRNA表达的影响。结果:SSTF能显著抑制HSC-T6细胞增殖、诱导细胞凋亡,且呈剂量和时间依赖;Western blotting结果显示,SSTF通过抑制Bcl-2,促进Bax和Caspase-3蛋白表达促进细胞凋亡;进一步采用Real-time PCR研究发现,Bcl-2能够从mRNA水平抑制Bcl-2、促进Bax和Caspase-3表达。结论:SSTF具有促进HSC-T6凋亡的作用,这种作用的发挥主要是通过抑制Bcl-2、促进Bax和Caspase-3蛋白和mRNA的表达实现的。Objective： To study the effect of total flavanones of Sedum sarmentosum （SSTF） on the apoptosis of rat hepatic stellate cells （HSC-T6） and its mechanism. Method： Different concentrations of SSTF and HSC-T6 cells were co-cultured for different period of time. The MTr assay was used to detect the inhibitory effect of SSTF on the proliferation of HSC-T6 cells. The flow cytometry An- nexin-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. Result： SSTF significantly inhibited HSC-T6 cell proliferation and induced cell apoptosis in a dose and time dependent manner. Ac- cording 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. Conclusion ： SSTF has the effect of promoting the apoptosis of HSC-T6 mainly by inhibiting Bcl-2 and promoting protein and mRNA ex- pressions of Bax and caspase-3.

关 键 词：垂盆草总黄酮 肝星状细胞 凋亡 

分 类 号：R285[医药卫生—中药学]