蛹虫草提取物对高糖诱导内皮细胞衰老的影响

摘要/Abstract

摘要： 目的: 探讨蛹虫草乙酸乙酯提取物(CME)对高糖诱导人脐静脉内皮细胞(HUVECs)衰老的影响。方法: 用高糖(40 mmol/L) 诱导建立HUVECs衰老体外模型,以冬虫夏草提取物(CSE)(50 μg/mL)为阳性对照,观察CME(12.5~100 μg/mL)对内皮细胞衰老的干预作用。四甲基偶氮唑蓝(MTT)比色法检测细胞增殖;SA-β-半乳糖酐酶(SA-β-gal)染色法检测细胞衰老特异性的SA-β-gal活性;流式细胞术检测细胞周期分布和细胞内活性氧(ROS)含量。结果: 经高糖干预24、48、72 h 后,HUVECs增殖活性OD值明显低于对照组;而且在高糖孵育 96 h 后,SA-β-gal染色阳性细胞率明显增加;孵育 48 h 后,处于G₀/G₁期细胞的百分率和ROS相对水平明显增加(P<0.01)。与高糖组相比,CME(25~50 μg/mL)孵育48和 72 h 后细胞OD值显著提高(P<0.05);CME(12.5~100 μg/mL)孵育 96 h 后SA-β-gal染色阳性细胞率显著降低(P<0.01);孵育 48 h 后G₀/G₁期的细胞百分率(P<0.01)、细胞内ROS水平显著降低(P<0.05);CME 25、50 μg/mL 效果优于CME 12.5、100 μg/mL(P<0.05),且与CSE 50 μg/mL 作用相近(P>0.05)。结论: 蛹虫草乙酸乙酯提取物可促进内皮细胞生长增殖,可能与其降低细胞内ROS水平,减轻细胞氧化应激损伤有关,从而减轻内皮细胞衰老。

关键词: 人脐静脉内皮细胞, 高糖, 细胞衰老, 抗氧化活性, 蛹虫草乙酸乙酯提取物

Abstract: AIM: To determine the effect of the ethyl acetate extract of cordyceps militaris (CME) on senescence in cultured human umbilical vein endothelial cells (HUVECs) treated with high glucose. METHODS: The HUVECs senescence model was established by incubation with high glucose(40 mmol/L) , the extract of Cordyceps Sinensis(CSE) (50 μg/mL) was treated as a positive control, and then CME (12.5-100 μg/mL) was treated to observe its effect on cell senescence. Cell proliferation was determined by the methylthiazolyl tetrazolium (MTT) reduction assay , SA-β-galactosidase (SA-β-gal) positive cells were detected by X-gal, and phase distribution of cell cycles and cellular reactive oxygen species (ROS) were detected by flow cytometry. RESULTS: Compared with unintervented cells, cell OD value significantly decreased in cells treated with high glucose for 24, 48, and 72 h (P<0.01). The percentage of cells positive for SA-β-gal, the percentage in G₀/G₁ phase and ROS levels notably increased after cells incubated by high glucose for 96 or 48 h (P<0.01). Compared with high glucose intervention cells, cell OD value significantly increased in cells intervented by CME (25-50 μg/mL) for 48 h and 72 h ( P<0.05); CME(12.5-100 μg/mL)stimulating for 96 h significantly lowered the percentage of cells positive for SA-β-gal (P<0.01); CME(12.5-100 μg/mL)incubation for 48 h significantly lowered the percentage of G₀/G₁ phase of HUVECs (P<0.01) and ROS production (P<0.05); All these effects of CME 25, 50 μg/mL groups were better than those12.5, 100 μg/mL groups (P<0.05), and were similar to CSE 50 μg/mL group(P>0.05). CONCLUSION: Ethyl acetate extract of cordyceps militaris may promote the HUVECs growth and proliferation, protecting from high glucose-stimulated vascular endothelial senescence. The effects may be associated with ethyl acetate extract of cordyceps militaris inhibiting reactive oxygen species and alleviating oxidative stress injury.

Key words: Human umbilical vein endothelial cells, High glucose, Cell senescence, Antioxidative activity, Ethyl acetate extract of cordyceps militaris

中图分类号:

R965

朱鹏立, 叶章正, 余惠珍, 黄华玮, 阮景明, 林帆. 蛹虫草提取物对高糖诱导内皮细胞衰老的影响[J]. 中国临床药理学与治疗学, 2012, 17(11): 1216-1222.

ZHU Peng-li, YE Zhang-zheng, YU Hui-zhen, HUANG Hua-wei, RUAN Jing-ming, LIN Fan. Effect of the extract of cordyceps militaris on senescence in cultured human umbilical vein endothelial cells treated with high glucose[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(11): 1216-1222.

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