丹参不同组分对大鼠血管平滑肌细胞增殖的影响

Influence of different component of danshen on the proliferation in vessel smooth muscle cell

关键词:  丹参  丹参酮  丹酚酸  血管平滑肌细胞  增殖  细胞周期

Key words:  Danshen  Danshen ketone  phenic acid  vessel smooth muscle cell(VSMC)  proliferation  cell cycle

摘要:

[目的]探讨丹参的水溶性有效成分(总丹酚酸)和脂溶性有效成分(总丹参酮)对大鼠血管平滑肌细胞(VSMC)增殖的影响,以寻找药物的作用靶点。[方法]分离大鼠主动脉中层平滑肌,贴壁法培养平滑肌细胞,无血清培养基培养静止后,以5%胎牛血清(FCS)刺激VSMC增殖,分别加入不同浓度的总丹参酮或丹酚酸,以四甲基偶氮唑蓝(MTT)法观察药物对VSMC增殖的影响;以Bradford法检测细胞总蛋白;流式细胞仪检测药物对细胞周期的影响。[结果]与对照组相比丹参酮使VSMC增殖活性下降(P<0.05),细胞总蛋白合成降低(P<0.05),并呈浓度依赖性,而丹酚酸组与对照组相比无显著性差异;此外流式细胞术检测结果显示丹参酮使VSMCG0/G1期比例升高,S期比例下降,而丹酚酸组与对照组相比仍无显著性差异。[结论]丹参的脂溶性有效成分总丹参酮具有抑制血管平滑肌细胞增殖的作用,而丹酚酸对VSMC不具有以上作用。

Abstract:

[Objective]To explore the influence of water-soluble active principle(total phenic acid)and fat-soluble active principle(danshen ketone)in Danshen on the proliferation of vessel smooth muscle cell(VSMC),and search for the target point.[Methods]Middle-layer smooth muscle of aorta in rats was separated,smooth muscle cell was cultured by stick-wall method.After cultured by no serum medium,proliferation of VSMC was stimulated by5%FCS,and then different con-centration of total phenic acid and ketone was added.The influence was observed by MTT method.The total protein was determined by Bradford method.Influence of herbs on cell cycle was detected by flow cytometry.[Result]Compared with the control group,the proliferation activity of VSMC was decreased(P<0.05)in Danshen ketone group and the synthesis of total protein descended(P<0.05)and depended on the concentration.There had no significantly difference between control group and phenic acid group.Otherwise,the proportion of VSMC G0/G1periods was ascended by Danshen ketone,the proportion of S periods descended.There had no significantly difference between control group and phenic acid group.[Conclusion]The fat-soluble active principle(total Danshen ketone)in Danshen possess inhibiting effect on the proliferation of VSMC,while phenic acid has no effect on VSMC like this.