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目的:观察人参三七川芎提取物对复制性衰老人心脏微血管内皮细胞(HCMEC)的影响和血管内皮钙黏蛋白(VE-cadherin)的作用。方法:以HCMEC为研究对象,第8代HCMEC建立复制性衰老HCMEC模型,实验分为衰老模型组(衰老组)、白藜芦醇组(10μmol·L-1)、人参三七川芎低、中、高剂量组(50,100,200 mg·L-1),相应药物作用24 h后,采用β-半乳糖苷酶(SA-β-gal)衰老染色鉴定,CCK-8检测细胞增殖能力,流式细胞仪分析细胞周期,采用激光免疫共聚焦显微镜观察VEcadherin的形态学改变,Western blot检测VE-cadherin蛋白的表达情况。结果:与衰老组比较,人参三七川芎提取物各剂量组减少SA-β-gal衰老染色比例,促进细胞的增殖,减少G0/G1期细胞比例,增加G2/M期细胞比例,保持VE-cadherin形态的完整性,升高VE-cadherin蛋白的表达,均具有明显的统计学差异(P<0.05,P<0.01)。结论:人参三七川芎提取物能够延缓衰老,促进增殖和有丝分裂,保持复制性衰老HCMEC VE-cadherin的完整性和蛋白的表达,这可能是益气活血中药延缓内皮衰老的重要机制所在。
Objective: To observe the effects of ginseng and rhizoma chuanxiong rhizome extract on human cardiac microvascular endothelial cells (HCMEC) and the effect of vascular endothelial cadherin (VE-cadherin) in replicating aged human. Methods: HCMEC was used as the research object to establish replicative senescent HCMEC model on the 8th generation of HCMEC. The experiment was divided into aging model group (aging group), resveratrol group (10μmol·L-1), ginseng , High dose group (50,100,200 mg · L-1) and corresponding drug for 24 h, then identified by SA-β-gal senescence staining. The proliferation of CCK-8 cells was detected by flow cytometry The cell cycle was analyzed. The morphological changes of VE cadherin were observed by laser confocal microscopy. The expression of VE-cadherin protein was detected by Western blot. Results: Compared with aging group, each dose group of Ginseng Extract of Chuanxiong notoginseng reduced the proportion of SA-β-gal aging staining, promoted cell proliferation, decreased the proportion of cells in G0 / G1 phase, increased the proportion of cells in G2 / M phase and maintained VE- cadherin morphological integrity, increased VE-cadherin protein expression, have significant statistical difference (P <0.05, P <0.01). Conclusion: The extract of Panax Chuanxiong could delay aging, promote proliferation and mitosis, maintain the integrity and protein expression of VE-cadherin, which may be the important mechanism of Yiqi Huoxue medicine in delaying endothelial aging.