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目的观察阿司匹林对脂多糖(LPS)诱导的体外培养大鼠主动脉平滑肌细胞增殖的影响。方法采用组织贴块法培养大鼠主动脉平滑肌细胞,选3-9代细胞用于实验。用乳酸脱氢酶活性测定法进行细胞毒性试验,观察阿司匹林加入对细胞有无毒性。采用四唑盐比色法计数不同浓度阿司匹林处理下,LPS诱导的大鼠血管平滑肌细胞增殖数量的变化。用流式细胞仪检测分别加入LPS和阿司匹林2mmol/L后大鼠血管平滑肌细胞周期变化。结果与对照组比较,阿司匹林5mmol/L组细胞上清液中乳酸脱氢酶活性无明显升高(P>0.05)。阿司匹林(0.5、1、2mmol/L)呈剂量依赖性的抑制大鼠主动脉平滑肌细胞的增值,实验显示2mmol/L浓度的阿司匹林在培养第五天时抑制的效果最大,与LPS组比较有极显著的差异。与LPS组比较,阿司匹林(2mmol/L)可使细胞静止期/DNA合成前期(Go/G1)的大鼠主动脉平滑肌细胞所占比例显著升高,而DNA合成期(S期)细胞所占比例显著降低。结论阿司匹林能呈剂量依赖性地抑制LPS诱导的大鼠主动脉平滑肌细胞的增殖,并将细胞抑制在G0/G1。
Objective To observe the effect of aspirin on the proliferation of rat aortic smooth muscle cells induced by lipopolysaccharide (LPS) in vitro. Methods Rat aorta smooth muscle cells were cultured by tissue patch method. Cells of passage 3-9 were selected for experiment. Lactate dehydrogenase activity assay cytotoxicity test to observe the addition of aspirin on the cells with or without toxicity. The tetrazolium salt colorimetry was used to measure the changes of LPS-induced rat vascular smooth muscle cell proliferation under different concentrations of aspirin. Flow Cytometry was used to detect the changes of vascular smooth muscle cells in rat after 2mmol / L LPS and aspirin respectively. Results Compared with the control group, lactate dehydrogenase activity in the supernatant of 5mmol / L aspirin group was not significantly increased (P> 0.05). Aspirin (0.5, 1, 2mmol / L) dose-dependently inhibited the proliferation of rat aortic smooth muscle cells. The results showed that aspirin at 2mmol / L had the most inhibitory effect on the fifth day of culture, compared with LPS group The difference. Compared with LPS group, aspirin (2mmol / L) significantly increased the proportion of rat aorta smooth muscle cells in quiescent phase / early stage of DNA synthesis (S G / G1), while cells in S phase The proportion is significantly reduced. Conclusion Aspirin can inhibit LPS-induced proliferation of rat aortic smooth muscle cells in a dose-dependent manner and inhibit the cells in G0 / G1.