复方丹参方对TNF-α刺激血管平滑肌细胞影响的蛋白质组学研究

Effects of Fufangdanshen on TNF-α stimulated vascular smooth muscle cells using proteomics

目的观察复方丹参方对TNF-α刺激血管平滑肌细胞的影响,在分子水平上探讨复方丹参方治疗动脉粥样硬化的作用机制。方法用TNF-α刺激的血管平滑肌细胞来建立体外平滑肌细胞功能失常的细胞模型。用MTT和流式细胞术测定血管平滑肌细胞的增殖活力,用双向电泳、图像分析、质谱鉴定等蛋白质组学技术测定复方丹参方对血管平滑肌细胞中有影响的蛋白质。结果复方丹参方抑制了TNF-α引起的血管平滑肌细胞的增殖,TNF-α作用血管平滑肌细胞后有16个蛋白质下调和24个蛋白质上调,复方丹参方作用血管平滑肌细胞后可使这些改变的蛋白质水平有所恢复。复方丹参方可以下调钙调蛋白依赖蛋白激酶、N-ras、基质金属蛋白酶9的水平,上调p53肿瘤抑制因子、周期素依赖激酶抑制因子p21的水平。结论复方丹参方通过抑制基质金属蛋白酶9的分泌和升高周期素依赖激酶抑制因子的水平进而抑制血管平滑肌细胞的增殖与迁移是其治疗动脉粥样硬化的可能机制。

Aim To investigate the effects of Fufangdanshen on TNF-α stimulated vascular smooth muscle cells （VSMC） and discover potential molecular mechanism to cure atherosclerosis at molecular level, nethotis TNF-α stimulated VSMC was used as dysfunction cell model; VSMC proliferation was detected with MTY and FACS; proteomic protocol involving 2-DE, image analysis and spectrometry detection were used to detectregulated protein by Fufangdanshen. Results Fufangdanshen inhibited expression of 16 proteins but promoted expression of 24 proteins in TNF-α stimulated VSMC. The expression of CaMKK and N-ras were decreased by Fufangdanshen, which farther decreased matrix metalloproteinase production; while cell cycle related protein p21 , p53 were increased by Fufangdanshen in TNF-α treated VSMC. Conclusion The inhi-bition of proliferation and migration of VSMC through decreased matrix metalloproteinase production and increased cell cycle related protein p21, p53 might be the mechanism of Fufangdanshen to cure atherosclerosis.