阿司匹林对高糖和高胰岛素诱导的血管平滑肌细胞增殖的影响

Effects of aspirin on the high concentration of glucose and insulin-induced proliferation of vascular smooth muscle cells

目的:探讨阿司匹林对高糖和高胰岛素诱导的大鼠血管平滑肌细胞(VSMCs)增殖的影响及相关机制。方法:组织贴块法培养原代大鼠胸主动脉平滑肌细胞,用高糖和高胰岛素诱导细胞增殖。实验设对照组、高糖和高胰岛素组(HGI)、HGI+阿司匹林(0.5,1.25,2.50mmol/L)组。采用四甲基偶氮唑盐(MTT)比色法测定VSMCs增殖;一氧化氮(NO)试剂盒检测培养基上清液NO含量;流式细胞仪检测细胞周期。结果:阿司匹林呈剂量依赖性地抑制高糖和高胰岛素诱导的动脉VSMCs增殖,最大抑制作用出现在2.50mmol/L阿司匹林培养5d时(P<0.01);阿司匹林(2.50mmol/L)干预可使高糖和高胰岛素导致下降的NO含量上升(P<0.01);与高糖和高胰岛素组比较,阿司匹林(2.50mmol/L)可使细胞静止期/DNA合成前期(G0/G1期)的VSMCs所占比例显著升高(P<0.05),而DNA合成期(S期)细胞所占比例显著降低(P<0.05)。结论:阿司匹林能呈剂量依赖性地抑制高糖和高胰岛素诱导的动脉VSMCs的增殖,抑制细胞在G0/G1期,促进动脉平滑肌细胞NO的产生。

Objective To study the effect and related mechanisms of aspirin on high concentration of glucose and insulin-induced vascular smooth muscle cells （VSMCs） proliferation. Methods The primary VSMCs were cultured by planting Wistar rat thoracic vascular smooth muscle pieces. The proliferation of VSMCs was induced by high concentration of glucose and insulin. The VSMCs were divided into the control group, high concentration of glucose and insulin group （ HGI）, HGI ＋ aspirin（0.5, 1.25, 2.50 mmol/L） groups . Methylthiazolyl tetrazolium （MTT） colorimetric assay was used to measure the VSMCs proliferation. The content of nitric oxide （NO） in cell culture medium was detected by NO kit. Flow cytometry was used to evaluate the cell cycle. Results The largest anti-proliferative effect on VSMCs of aspirin was 2.50 mmoL/L concentration on the fifth day in this experiment, compared with the HGI, a very significant inhibition （P 〈0.01 ） ; HGI ＋ aspirin （2.50 mmol/L） group can be very significantly increased the content of NO that declined by high concentration of glucose and insulin in cell culture medium （P 〈0.01 ） ; Compared with the HGI group, aspirin （2.50 mmol/L） can increase the proportion of G0/G1 of VSMCs significantly （P 〈 0.05 ）, while the period of DNA synthesis （ S phase） cells in a significant reduction in the proportion （P 〈 0.05）. Conclusion Aspirin can dose-dependently inhibit high concentration of glucose and insulin-induced VSMCs proliferation, the point of inhibition is at the G0/G1 phase, and promote the NO production of VSMCs.