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目的:观察淫羊藿苷对高糖环境下内皮细胞衰老的影响及可能的作用机制研究。方法:人脐静脉内皮细胞分别培养在正常糖浓度(5.5 mmol/L),高糖浓度(33 mmol/L),高糖+淫羊藿苷低剂量及高糖+淫羊藿苷高剂量组中,药物干预48 h后,β半乳糖苷酶染色测定细胞衰老,流式细胞术检测细胞周期,NO检测试剂盒检测细胞培养上清中NO水平,Westernblot检测培养细胞PI3K,Akt蛋白表达水平。结果:与正常糖浓度组比较,高糖浓度组衰老细胞的阳性率明显增高,G0/G1期细胞比率增加,S期显著减少,细胞培养上清中NO水平减少,PI3K和磷酸化Akt蛋白水平降低。而淫羊藿苷能够明显降低高糖环境下衰老细胞阳性率,增加S期细胞比率,提高培养上清中NO水平,同时能够提高高糖环境下内皮细胞PI3K和磷酸化Akt蛋白表达水平。结论:淫羊藿苷能延缓高糖诱导的内皮细胞衰老进程,提高内皮细胞功能,其作用机制可能与调节高糖环境下内皮细胞PI3k/Akt通路有关。
Objective: To observe the effect of icariin on the endothelial cell senescence under high glucose and its possible mechanism. Methods: Human umbilical vein endothelial cells were cultured in high glucose (5.5 mmol / L), high glucose (33 mmol / L), high glucose + icariin low dose and high glucose + icariin high dose The cell cycle was determined by β-galactosidase staining, the cell cycle was detected by flow cytometry, the level of NO in cell culture supernatant was detected by NO assay kit, and the protein expression levels of PI3K and Akt in cultured cells were detected by Western blot. Results: Compared with the normal glucose concentration group, the positive rate of senescent cells in high glucose group was significantly increased, the ratio of cells in G0 / G1 phase increased, the S phase decreased significantly, the level of NO in cell culture supernatant decreased, the levels of PI3K and phosphorylated Akt protein reduce. Icariin could significantly decrease the positive rate of senescence cells in high glucose, increase the ratio of S phase cells, increase the level of NO in culture supernatant, and increase the expression of PI3K and phospho-Akt in endothelial cells under high glucose. CONCLUSION: Icariin can delay the process of endothelial cell senescence induced by high glucose and enhance the function of endothelial cells. Its mechanism may be related to the regulation of PI3k / Akt pathway in endothelial cells under high glucose condition.