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

doi:10.3969/j.issn.1004-616x.2017.01.003

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (1): 13-17,22.doi: 10.3969/j.issn.1004-616x.2017.01.003

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

由淑萍¹, 赵军², 马龙¹, 张石蕾¹, 刘涛¹

1. 新疆医科大学公共卫生学院卫生毒理学教研室, 新疆乌鲁木齐 830011;
2. 新疆维吾尔族自治区药物研究所, 新疆乌鲁木齐 830004

收稿日期:2016-04-18 修回日期:2016-06-24 出版日期:2017-01-31 发布日期:2017-01-31
通讯作者: 刘涛,E-mail:xjmult@163.com;Tel:18997956799 E-mail:xjmult@163.com
作者简介:由淑萍,E-mail:youshupin@163.com
基金资助:
国家自然科学基金项目（81260624）

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

YOU Shuping¹, ZHAO Jun², MA Long¹, ZHANG Shilei¹, LIU Tao¹

1. Department of Toxicology, College of Public Health, Xinjiang Medical University, Urumqi 830011;
2. Xinjiang Uygur Autonomous Region Institute of Materia Medica, Urumqi 830004, Xinjiang, China

Received:2016-04-18 Revised:2016-06-24 Online:2017-01-31 Published:2017-01-31

摘要/Abstract

摘要：

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。

关键词: 肉苁蓉苯乙醇总苷, 毛蕊花糖苷, 松果菊苷, 肝星状细胞, 凋亡

Abstract:

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.

Key words: cistanche phenylethanoid glycosides, acteoside, echinacoside, hepatic stellate cells, apoptosis

中图分类号:

由淑萍, 赵军, 马龙, 张石蕾, 刘涛. 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[J]. 癌变·畸变·突变, 2017, 29(1): 13-17,22.

YOU Shuping, ZHAO Jun, MA Long, ZHANG Shilei, LIU Tao. Effects of cistanche phenylethanoid glycosides, acteoside and echinacoside on inhibition of cell proliferation and inducation of apoptosis in rat hepatic stellate cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(1): 13-17,22.

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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

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

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