丹参抗成纤维细胞增殖作用的分子机制研究  

Antiproliferative molecular mechanisms of components in roots and rhizomes of Salvia miltiorrhiza on fibroblasts

作　　者：张楠楠[1] 王倩[2] 高洁[3] 李红艳 韩彦琪[3] 邹敏杰 白钢[3] 

机构地区：[1]天津中医药大学,天津300193 [2]河南广播电视大学,河南郑州450008 [3]南开大学药学院,天津300071 [4]天津嘉氏堂科技有限公司,天津300457

出　　处：《中草药》2012年第12期2440-2447,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81173638;81102835;81001682)

摘　　要：目的探讨丹参抗成纤维细胞增殖作用的有效成分及分子作用机制。方法采用索氏提取法制备丹参多种提取物及萃取部位;以抗人增生性瘢痕成纤维细胞(HSF)增殖的活性为导向,MTS法筛选活性部位,流式细胞仪检测细胞周期;UPLC-Q/TOF法分析丹参活性成分;通过反向对接预测试药抗细胞增殖的机制,RT-PCR检测caspase-3、MAP2K1、MAPK1增殖相关基因的表达水平。结果丹参石油醚提取部位有较强的抗HSF增殖的活性,能显著增加G0/G1期细胞的比例,降低增殖指数(PI),其主要活性成分为丹参酮类化合物;反向对接预测结果显示,丹参酮类化合物抗HSF增殖机制主要与调节caspase-3和MAP2K1等靶点及其相关通路有关;隐丹参酮能够上调caspase-3基因的表达,下调MAP2K1、MAPK1基因的表达,与预测结果相吻合。结论丹参抗HSF增殖作用的机制与丹参酮类成分调节MAPK、VEGF等信号通路有关。Objective To investigate the active components in the roots and rhizomes of Salvia miltiorrhiza and their antiproliferative molecular mechanisms on fibroblasts.Methods Different components and extraction fractions from the roots and rhizomes of S.miltiorrhiza were prepared using Soxhlet extraction method.MTS method was used to screen the active components guided by the antiproliferative activity on human hypertrophic scar fibroblasts（HSF）,flow cytometry was used to detect their influence on cell cycle,and the active components were analyzed by UPLC-Q/TOF method.The antiproliferative mechanism was predicted by reverse docking,and the gene expression levels of caspase-3,MAP2K1,and MAPK1 were determined using RT-PCR technology.Results The petroleum ether extract fraction from the roots and rhizomes of S.miltiorrhiza had higher antiproliferative activity on human HSF,increased the proportion of cells in G0/G1 phase significantly,and reduced the proliferation index（PI）.The main active components were tanshinones.Results of reverse docking indicated the antiproliferative mechanisms on human HSF,might relate to the regulation of some target proteins,such as caspase-3 and MAP2K1,and the related pathways.RT-PCR results showed that cryptotanshinone could upregulate the gene expression of caspase-3 and downregulate the gene expression of MAP2K1 and MAPK1.Conclusion The antiproliferative mechanisms of the components in the roots and rhizomes of S.miltiorrhiza on human HSF may be related with the regulation of MAPK and VEGF signaling pathways by tanshinone components.

关 键 词：丹参 人增生性瘢痕成纤维细胞 MAPK1 反向对接 丹参酮 

分 类 号：R282.710.5[医药卫生—中药学]