黄酒多酚对大鼠血管平滑肌原代细胞增殖和迁移的影响及其机制研究

THE EFFECT AND MECHANISM OF YELLOW WINE POLYPHENOL COMPOUNDS ON VSMCS PROLIFERATION AND MIGRATION

目的探讨黄酒多酚（ YWPC）对同型半胱氨酸（ Hey）诱导的大鼠血管平滑肌细胞（ VSMCs ）增殖和迁移的影响及其可能的作用机制.方法： SD 大鼠胸主动脉VSMCs原代细胞培养鉴定,取第 4-7 代细胞用于实验.VSMCs分为对照组、 Hcy组、 YWPC1 mg/L 组 、YWPC 10 mg/L 组及 YWPC 100 mg /L 组.MTT 法测定各组VSMCs的增殖情况;划痕法和Tanswell法 分别检测VSMCs的迁移、侵袭情况;采用 Westrnblotting 法检测细胞周期相关蛋白P21、 P27、基质金属蛋白酶（ MMP ） -2、 MMP-9、哺乳动物雷帕霉素靶蛋白（ mTOR ）、磷酸化（p-）mTOR、 70ku 核糖体蛋白S6 激酶（ P70S6K）、 p-P70S6K的表达情况; 明胶酶谱法检测MMP-2、 MMP -9的活性.结果：与对照组比较, Hcy组 VSMCs增殖和迁移增加（P〈0.05 ）MMP - 2、 MMP- 9、 p-mTOR、 p-P70S6K 表达量增加（P〈0.05） ;P21、 P27 表达量减少（P〈0. 05 ）.与Hcy 组比较, YWPC 组VSMCs增殖和迁移减弱（P〈0. 05 或P〈0. 01） ;MMP-2、 MMP-9、 p-mTOR、 p-P70S6K 表达量减少（P〈0. 05 或 P〈0. 01） ; P21、 P27 表达量增加（P〈0. 05 ）.结论： YWPC能够抑制Hcy诱导的VSMCs增殖和迁移,其机制可能与抑制mTOR /P70S6K信号通路有关.

Objective： To investigate the effect and mechanism of yellow wine polyphenol eompounds （YW- PC） on the homocysteine （Hcy.）-induced rat aortic vascular smooth muscle eells （VSMCs） proliferation and migration. Methods： Primary SD rat VSMCs were generated with tissue fragment culture method,and 4 - 7th generations of cells were used for further experiments. The cells were divided into 5 groups： the con-trol group,the Hcy group,and low- （1 mg/L） ,medium- （10 mg/L） and high-dose （100 mg/L） YWPC- treated groups. The proliferation of VSMCs was measured by MTT assay. The migration and invasion of VSMCs were detected by wound healing and trans-well invasion assay respectively. The expression of P21, P27,matrix metallo preteinases （MMP）-2, MMP-9,mammalian target of rapamycin （mTOR）, p-mTOR , 70 ku ribosomal protein S6 kinase （P70S6K） and p-P70S6K in VSMCs were determined by western blot-ting. The enzymatic activities of MMP-2 and MMP-9 were detected by gelatin zymography. Results： Com-pared with the control group,the proliferation and migration of VSMCs as well as the expression levels of MMP-2,MMP-9,p-mTOR and p-P70S6K were increased,whiee the P21 and P27 expression levels were decreased in the Hcy group （ P 〈 0 . 05）. The proliferation and migration of VSMCs were attenuated and the expression of MMP-2, MMP-9, p-mTOR and p-P70S6K were down-regulated, whereas the P21 and P27 expression were up-regulated in the YWPC-treated groups compared to the Hcy group （ P 〈0. 05）.Conclusion ： YWPC could suppress Hcy-induced VSMCs proliferation and migration, possibly via the inhibition of the mTOR/P70S6K signaling pathway.