香青兰总黄酮对TNF-α诱导的大鼠血管平滑肌细胞增殖的影响

Inhibitory Effects of Dracocephalum Total Flavoes on Proliferation of Rat Vascular Smooth Muscle Cells Induced by TNF-α

目的:通过观察香青兰总黄酮对肿瘤坏死因子-α(TNF-α)诱导的大鼠主动脉血管平滑肌细胞(VSMC)增殖的影响,探讨其治疗动脉粥样硬化的可能药效机制。方法:采用体外贴块法培养大鼠主动脉VSMC,SABC-Cy3荧光免疫组化技术鉴定其纯度;分别应用质量浓度为0.02 mg.L-1 TNF-α,TNF-α+不同浓度香青兰总黄酮组(25,50,100 mg.L-1)作用24 h,并设对照组进行比较。采用MTT法检测细胞的增殖;免疫组化法检测细胞内增殖细胞核抗原(PCNA)的表达水平。结果:与对照组比较,TNF-α组能显著刺激大鼠VSMC的增殖,香青兰总黄酮不同剂量组联合TNF-α可一定程度抑制TNF-α诱导的VSMC增殖以及细胞内PCNA的表达,且呈现一定的剂量依赖关系趋势。结论:香青兰总黄酮通过抑制TNF-α对VSMC的增殖作用,可能是其治疗动脉粥样硬化的药效机制之一。

Objective：To explore the possible mechanism of pharmacological activity of tilianin by observing the effects of Dracocephalum total flavoes on proliferation of rat vascular smooth muscle cells（VSMCs）induced by TNF-α.Method： The rat VSMCs were cultured in vitro,and the purity of cells was identified by SABC-Cy3 immunofluroescence technique.Cell proliferation model was established by stimulation with 0.02 mg·L-1 TNF-α.After TNF-α stimulation,cells were treated with three dose of Dracocephalum total flavoes（25,50,100 mg·L-1）;MTT assay was used to evaluate the proliferation of cells.The expression of intracellular proliferating cell nuclear antigen（PCNA）were measured by immunoenzymatic histochemistry.Result： Compared with control group,three dose groups of Dracocephalum total flavoes could inhibit,in varying degrees,the proliferation and expression of intracellular PCNA of VSMCs induced by TNF-α,in a dose-dependent manner.Conclusion：The Dracocephalum total flavoes could inhibit the proliferation of VSMC induced by TNF-α,which might be one of pharmacological mechanisms of Dracocephalum total flavoes in treatment for atherosclerosis.