OBJECTIVE:To study tanshinone ⅡA inhibiting the proliferation and migration induced by homocysteine(Hcy)of rat aortic vascular smooth muscle cells(VSMCs)and its signal pathway. METHODS:VSMCs were selected for the following ex-periments. In order to validate TanshinoneⅡA inhibiting the proliferation and migration induced by Hcy of VSMCs,VSMCs were di-vided into control group,Hcy group(1 000μmol/L),TanshinoneⅡA low-dose,medium-dose and high-dose groups(5,10,20 µg/L). In mechanism study test,VSMCs were divided into control group,TanshinoneⅡA group(20 µg/L),rapamycin group(20 nmol/L), MHY1485 group(10μmol/L),TanshinoneⅡA+rapamycin group(TanshinoneⅡA,20 µg/L+rapamycin,20 nmol/L),TanshinoneⅡA+MHY1485 group (TanshinoneⅡA,20 µg/L+MHY1485,10 μmol/L). In validation test of P70S6K and p-P70S6K pathway expres-sion,rapamycin and MHY1485 were used for inhibitory test and activation test,respectively. The proliferation of VSMCs was de-termined by ELIASA,and Transwell chambers and wound healing test were employed to test the migratory ability of VSMCs. West-ern blotting were used to investigate the expressions of P21,P27,MMP-2,MMP-9,P70S6K and p-P70S6K in VSMCs. RE-SULTS:In validation test,compared with control group,24,48 h absorbance,migration area,the number of VSMCs penetration and the expression of MMP-2,MMP-9 and p-P70S6K increased significantly,while the expression of P21 and P27 decreased sig-nificantly(P<0.01). Compared with Hcy group,48 h absorbance of TanshinoneⅡA low-dose group,24,48 h absorbance and the expression of MMP-2 of TanshinoneⅡA medium-dose and high-dose groups,migration area,the number of VSMCs penetration, the expression of MMP-9 and p-P70S6K in low-dose,medium-dose and high-dose groups all decreased significantly;the expres-sion of P21 and P27 increased significantly in TanshinoneⅡA medium-dose and high-dose groups(P<0.01);there was no statisti-rapamycin group,while 24,48 h absorbance increased significantly in MHY1485 group(P<0.01). Compared with TanshinoneⅡA group,24,48 h absorbance decreased significantly in TanshinoneⅡA+rapamycin group,while 12,24,48 h absorbance,migration area and the number of VSMCs penetration increased significantly in TanshinoneⅡA+MHY1485 group (P<0.01). In inhibitory test,compared with control group,the expression of p-P70S6K decreased significantly in TanshinoneⅡA group (P<0.01);com-pared with TanshinoneⅡA group,the expression of p-P70S6K decreased significantly in rapamycin group and TanshinoneⅡA+rapa-mycin group(P<0.01);in activation test,the expression of p-P70S6K decreased significantly in TanshinoneⅡA group(P<0.01), compared with TanshinoneⅡA group,the expression of p-P70S6K increased significantly in MHY1485,TanshinoneⅡA+MHY1485 group (P<0.01). CONCLUSIONS:TanshinoneⅡA can inhibit the proliferation and migration of VSMCs by suppressing mTOR/P70S6K signal pathway.%目的：研究丹参酮ⅡA抑制同型半胱氨酸（Hcy）诱导的大鼠主动脉血管平滑肌细胞（VSMCs）增殖和迁移及其机制。方法：取VSMCs用于试验。在验证丹参酮ⅡA抑制Hcy诱导的VSMCs增殖和迁移试验（验证试验）中，将细胞分为对照组、Hcy组（1000μmol/L）及丹参酮ⅡA低、中、高剂量组（5、10、20µg/L）。在丹参酮作用机制试验（细胞通路试验）中，将细胞分为对照组、丹参酮ⅡA组（20µg/L）、雷帕霉素组（20 nmol/L）、MHY1485组（10μmol/L）、丹参酮ⅡA+雷帕霉素组（丹参酮ⅡA，20µg/L+雷帕霉素，20 nmol/L）、丹参酮ⅡA+MHY1485组（丹参酮ⅡA，20µg/L+MHY1485，10μmol/L）。验证细胞通路P70S6K和p-P70S6K的表达时，雷帕霉素（抑制试验）与MHY1485（激动试验）分别进行试验。酶标仪测定VSMCs的增殖，细胞划痕试验、Transwell法测定VSMCs的迁移，增强化学发光法测定VSMCs中P21、P27、基质金属蛋白酶（MMP）-2、MMP-9、P70S6K和p-P70S6K的表达。结果：验证试验中，与对照组比较，Hcy组细胞24、48 h吸光度值，迁移面积和VSMCs穿透数量，MMP-2、MMP-9和p-P70S6K表达水平显著升高，P21、P27表达水平显著降低（P＜0.01）；与Hcy组比较，丹参酮ⅡA低剂量组细胞48 h，中、高剂量组24、48 h吸光度值， MMP-2水平，低、中、高剂量组细胞迁移面积和VSMCs穿透数量，MMP-9和p-P70S6K表达水平显著降低，丹参酮ⅡA中、高剂量组细胞中P21、P27表达水平显著升高（P＜0.01）；各组细胞P70S6K水平比较，差异无统计学意义（P＞0.05）。细胞通路试验中，与对照组比较，丹参酮ⅡA、雷帕霉素组细胞24、48 h吸光度值，迁移面积和VSMCs穿透数量显著降低，MHY1485组24、48 h吸光度值显著升高（P＜0.01）；与丹参酮ⅡA组比较，丹参酮ⅡA+雷帕霉素组细胞24、48 h吸光度值显著降低，丹参酮ⅡA+MHY1485组细胞12、24、48 h吸光度值，迁移面积和VSMCs穿透数量显著升高（P＜0.01）。抑制试验中，与对照组比较，丹参酮ⅡA组细胞p-P70S6K表达水平显著降低（P＜0.01）；与丹参酮ⅡA组比较、雷帕霉素、丹参酮ⅡA+雷帕霉素组细胞p-P70S6K表达水平显著降低（P＜0.01）。激动试验中，丹参酮ⅡA组细胞p-P70S6K水平显著降低（P＜0.01）；与丹参酮ⅡA组比较，MHY1485、丹参酮ⅡA+MYH1485组细胞p-P70S6K水平显著升高（P＜0.01）。结论：丹参酮ⅡA可以抑制VSMCs的增殖和迁移，并通过抑制mTOR/P70S6K信号通路而发挥作用。
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