藏药波棱瓜子总木脂素对大鼠肝星状细胞增殖与凋亡的影响  

Effects of total lignans from Tibetan medicine Herpetospermum pedunculosum seeds on proliferation and apoptosis of rat hepatic stellate cells

作　　者：刘伟 石林琳 石晏丞 巴珂 李佳川[1] 顾健[1] 

机构地区：[1]西南民族大学药学院,四川成都610000

出　　处：《中草药》2017年第14期2912-2917,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81274168;81573563);四川省中医药管理局科研项目(2016C062)

摘　　要：目的通过研究藏药波棱瓜子总木脂素(TLHPS)对大鼠肝星状HSC-T6细胞的增殖抑制作用及诱导其凋亡的作用,进一步探讨TLHPS抗肝纤维化的作用机制。方法实验分为对照组,TLHPS 10、20、30、40μg/mL组,阳性药秋水仙碱0.1μg/mL组。用加入各组药物的培养基分别培养HSC-T6细胞24、48、72 h,采用CCK8法检测各组细胞24、48、72 h的增殖抑制率,流式细胞术检测细胞凋亡率及细胞周期,Western blotting法检测凋亡蛋白Bcl-2、Bax、核转录因子-κB(NF-κB)的表达水平。结果与对照组相比,TLHPS 10、20、30、40μg/mL组对HSC-T6细胞24、48、72 h时的增殖抑制率明显增高(P<0.01),且各组HSC-T6细胞早期凋亡率与晚期凋亡率明显升高(P<0.05,0.01),G0/G1期细胞数明显增多(P<0.01),G2/M期细胞数无明显差异(P>0.05),S期细胞数明显减少(P<0.01)。Western blotting结果显示,与对照组相比,TLHPS10、20、30、40μg/mL组Bcl-2蛋白表达明显降低(P<0.05,0.01),TLHPS 40μg/mL组Bax蛋白表达明显升高(P<0.01),TLHPS 20、30、40μg/mL组NF-κB表达明显降低(P<0.01)。结论 TLHPS抗肝纤维化的作用机制可能是通过降低Bcl-2、NF-κB的表达抑制肝星状细胞增殖并诱导其凋亡。Objective To observe the possible mechanism of total lignans from Herpetospermum pedunculosum seeds（TLHPS） in fighting against hepatic fibrosis of hepatic stellate cell-T6（HSC-T6） by studying the effects of TLHPS on inhibiting proliferation and inducing apoptosis of HSC-T6. Methods The cells were divided into totally six groups such as control group, TLHPS groups（10, 20, 30, 40 μg/mL）, and colchicine positive control group（0.1 μg/mL）. HSC-T6 cells were cultivated for 24, 48, and 72 h by six groups of culture medium containing different drugs. The proliferation inhibitory rate of HSC-T6 cells was detected by CCK8 method at 24, 48, and 72 h respectively. The apoptotic rate and cell cycle were detected by flow cytometry. Protein expression levels of B cell lymphoma-2（Bcl-2）, Bcl-2-associated X protein（Bax） and transcription factors-κB（NF-κB） were detected by Western blotting. Results Compared with control group, 24, 48, and 72 h proliferation inhibitory rate of HSC-T6 was obviously elevated in TLHPS 10, 20, 30, 40 μg/mL groups（P〈0.01）. Early-and late-stage apoptosis rates of HSC-T6 cells obviously increased in TLHPS 10, 20, 30, 40 μg/mL groups（P〈0.05, 0.01）. The cell numbers in G0/G1 phase were obviously increased in TLHPS groups（P〈0.01） and cell numbers in G2/M phase have no significant difference（P〈0.05）, cell numbers in S phase obviously decreased（P〈0.01）. Compared with control group, Bal-2 protein expression obviously decreased in TLHPS 10, 20, 30, 40 μg/mL groups（P〈0.05, 0.01）, Bax protein expression obviously increased in TLHPS 40 μg/mL group（P〈0.01） and NF-κB protein expression obviously decreased in TLHPS 20, 30, 40 μg/mL groups（P〈0.01）. Conclusion The mechanism of total lignans from Tibetan medicine Herpetospermum fighting against hepatic fibrosis might be associated with inhibiting proliferation and inducing apoptosis of HSC-T6. The inhibition of hepatic stellate cells proliferation and induction of its apoptosis may be r

关 键 词：波棱瓜子总木脂素 大鼠肝星状细胞 增殖 凋亡 BCL-2 BAX 核转录因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]