摘要
【目的】观察柴胡皂苷d对大鼠肝星状细胞内的雌激素受体转录激活的调节作用,探讨柴胡皂苷d的药理机制。【方法】体外培养大鼠肝星状细胞系HSC-T6,将雌激素受体的特异性报告基因ERE-tk-Luc用人工细胞膜介导细胞瞬时转染法转入HSC-T6细胞中,在不同药物浓度、不同时间作用点及加用雌激素受体抑制剂ICI182.780等条件下,采用双荧光素酶报告基因检测系统观察柴胡皂苷d及雌二醇对转染细胞荧光素酶(Luc)表达的影响。【结果】柴胡皂苷d浓度为0.01~5μmol/L、雌二醇浓度为0.01~1μmol/L时,荧光素酶活性呈剂量依赖性递增;柴胡皂苷d为5μmol/L,而雌二醇为1μmol/L时,荧光素酶活性最高;当雌二醇浓度到达5μmol/L时,效应减弱。且以5μmol/L柴胡皂苷D、1μmol/L雌二醇分别诱导细胞24 h时,荧光素酶表达活性最高。ICI182.780可明显抑制柴胡皂苷d、雌二醇对荧光素酶活性的诱导作用。【结论】柴胡皂苷d可促进HSC-T6细胞内雌激素受体转录激活。
Objective To observe the effect of saikosaponin-d on transcriptional activation of estrogen receptor in rat hepatic stellate cells（HSC-T6）, and to explore its pharmacological mechanism. Methods The rat HSC-T6 were cultured in vitro for the study. After transient transfection of HSC-T6 with the ER-specific reporter gene ERE-tk-Luc by liposome, the effect of saikosaponin-d and estradiol on luciferase activity was observed by Dual-Luciferase Reporter Assay System under the conditions of various drug concentrations, various treatment time and addition of estrogen receptor inhibitor ICI182. 780. Results When the concentrations of saikosaponin-d and estradiol were in the range of 0.01-5 μmol/L, 0.01-1 μmol/L respectively, luciferase activity was increased in a dose-dependent manner . Luciferase activity arrived the highest when saikosaponin-d concentration was 5 μmol/L and estradiol concentration was 1 μmol/L, but the effect was weakening when estradiol concentration reached 5μmol/L. In respect of the effect of treatment time, when HSC-T6 were separately treated with 5μmol/L of saikosaponin-d and 1 μmol/L of estradiol for 24 h, the luciferase activity was the highest. And ICI182.780 could significantly inhibit the induction of saikosaponin-d and estradiol for luciferase activity. Conclusion Saikosaponin-d has an effect on promoting the transcriptional activation of estrogen receptor in HSC-T6.
出处
《广州中医药大学学报》
CAS
2017年第4期550-555,共6页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(编号:81573775)
上海市卫生系统百人计划基金(编号:XBR2013120)
