丹参酮Ⅱ_A对血管平滑肌细胞内质网应激相关基因表达的影响

Effect of Tanshinone Ⅱ_A on Expression of Genes Related to Vascular Smooth Muscle Cells Endoplasmic Reticulum Stress

目的:研究丹参酮ⅡA对同型半胱氨酸(HCY)诱导增殖血管平滑肌细胞(VSMC)内质网应激(ERS)相关基因免疫球蛋白重链结合蛋白(Bip)表达的影响,探讨丹参酮ⅡA抗动脉粥样硬化的机制。方法:建立HCY诱导的兔VSMC增殖模型,与不同浓度的丹参酮ⅡA共同培养,Annexin/PI双染法、原位缺口末端标记(TUNEL)法检测细胞凋亡率,观察凋亡细胞形态;实时荧光定量PCR检测Bip基因表达。结果:丹参酮ⅡA可显著促进兔VSMC凋亡,细胞凋亡率与HCY组相比差异显著(P<0.01),且存在剂量依赖性;丹参酮ⅡA组Bip mRNA表达水平显著升高,与HCY组相比差异显著(P<0.05,P<0.01)。结论:丹参酮ⅡA上调Bip基因表达,放大ERS信号,诱导兔VSMC凋亡,可能是其抗动脉粥样硬化的机制之一。

Objective： Effect of tanshinone ⅡAon homocysteine（HCY） induced proliferation of vascular smooth muscle cells（VSMC） of endoplasmic reticulum stress（ERS） gene of immunoglobulin heavy chain binding protein（Bip） expression,to explore the mechanism of tanshinone ⅡA in atherosclerosis.Method： To establish the proliferation of rabbit VSMC model induced by HCY,incubated with different concentration of tanshinone ⅡA,Annexin / PI double staining,in situ nick end labeling（TUNEL） method to detect apoptosis rate,to observe the morphology of cell apoptosis;real-time fluorescent quantitative PCR detection the expression of Bip gene.Result： Tanshinone ⅡA can significantly promote the apoptosis of rabbit VSMC,cell apoptosis rate compared with HCY group（P 0.01）,and there was a dose dependent;expression levels of tanshinone ⅡA group Bip mRNA were significantly increased,compared with the HCY group had significant difference（P 0.05,P 0.01）.Conclusion： Expression of tanshinone ⅡA up-regulation of Bip gene,amplified ERS signal,induce apoptosis of rabbit VSMC,which may be one of the mechanisms of atherosclerosis.