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.