肉苁蓉苯乙醇总苷及其单体对肝星状细胞增殖和凋亡的影响

目的：探讨肉苁蓉苯乙醇总苷（CPhGs）及其单体（毛蕊花糖苷和松果菊苷）对大鼠肝星状细胞（HSC-T6）增殖及凋亡的影响。方法：分别以含不同浓度（0、3.91、7.81、15.63、31.25、62.50、125.00、250.00和500.00 μg/mL）CPhGs、毛蕊花糖苷及松果菊苷的DMEM（高糖）完全培养液处理HSC-T6细胞48 h，用四甲基偶氮唑盐（MTT）法检测细胞增殖情况并分别获得半数抑制浓度（IC₅₀）；再分别用不同浓度受试物诱导HSC-T6细胞48 h，试验分为正常对照组，CPhGs（25、50、100 μg/mL）组，毛蕊花糖苷（1.5、3、6.0 μg/mL）组及松果菊苷（125、250、500 μg/mL）组，流式细胞仪检测细胞凋亡的比例，Western blot检测细胞中凋亡蛋白Bcl-2和Bax的表达。结果：CPhGs、毛蕊花糖苷及松果菊苷对HSC-T6细胞的增殖抑制率随着浓度的增加逐渐升高，IC₅₀分别为119.1、7.0和520.3 μg/mL；与正常对照组比较，3种受试物诱导HSC-T6细胞的凋亡率差异均具有统计学意义（P < 0.01），其中，CPhGs 100 μg/mL、毛蕊花糖苷6.0 μg/mL及松果菊苷500 μg/mL浓度组的凋亡率分别为24.40%±3.16%，34.73%±1.16%和21.13%±0.98%；除毛蕊花糖苷1.5 μg/mL组外，其余不同浓度的受试物组均可引起HSC-T6细胞中Bax蛋白表达上调，Bcl-2蛋白表达下调（Bcl-2/Bax比值降低）（P < 0.05）。结论：CPhGs及其单体（毛蕊花糖苷和松果菊苷）可诱导HSC-T6细胞凋亡而抑制其增殖，从而起到抗肝纤维化的作用，3种受试物的抗肝纤维化效果排序为毛蕊花糖苷 > CPhGs > 松果菊苷。

Effects of cistanche phenylethanoid glycosides,acteoside and echinacoside on inhibition of cell proliferation and inducation of apoptosis in rat hepatic stellate cells

OBJECTIVE: To explore the effects of cistanche phenylethanoid glycosides (CPhGs),acteoside and echinacoside on anti-proliferation and induction of apoptosis in hepatic stellate cells (HSC-T6) in vitro. METHODS: HSC-T6 cells were treated with different concentrations of CPhGs (25,50,100 μg/mL),acteoside (1.5,3,6.0 μg/mL) or echinacoside (125,250,500 μg/mL) for 48 h. The MTT methods was used to measure the anti-proliferation effect and half the inhibition rate (IC₅₀) of HSC-T6. Proportions of apoptosis were assayed by flow cytometry. Western blot was used to detect the effects of the expression of Bcl-2 and Bax. RESULTS: With increasing concentrations of the three chemicals,the rate of inhibition of proliferation increased gradually. IC₅₀ were 119.1,7.0 and 520.3 μg/mL for the three chemicals,respectively. The proportions of apoptosis for the three chemicals were significantly different from that of the untreated controls (P < 0.01). The highest apoptosis rates for CPhGs (at 100 μg/mL),acteoside (6.0 μg/mL) and echinacoside (500 μg/mL) were 24.40%±3.16%,34.73%±1.16% and 21.13%±0.98%,respectively. Except for the 1.5 μg/mL acteoside group,expression of Bcl-2/Bax was significantly decreased (P < 0.05) compared to the normal control group. CONCLUSION: CPhGs,echinacoside and acteoside caused proliferation inhibition and apoptosis. The sequence of the hepato-protective effects of the three chemicals was shown to be acteoside > CPhGs > echinacoside.