过氧化氢对血管平滑肌细胞的作用

Effect of hydrogen peroxide to vascular smooth muscular cells

目的 研究过氧化氢 (H2 O2 )对兔血管平滑肌细胞的作用机制。方法 兔血管平滑肌细胞 (VSMCs)原代培养 ,微量细胞培养四甲基偶氮唑蓝法 (MTT)检测不同浓度H2 O2 在不同时段对其增殖影响 ,确定进一步研究的H2 O2 浓度 ,并应用流式细胞仪检测此浓度H2 O2 对VSMCs细胞周期和细胞凋亡的作用。结果 MTT法检测 ,H2 O2 浓度≥ 10 0 μmol/L(10 0～ 14 0 0 μmol/L)时 ,与VSMCs作用仅 1h ,即出现吸光度值显著降低 (P <0 .0 1) ;流式细胞仪细胞周期检测 ,H2 O2 浓度≥ 10 0 μmol/L时 ,与VSMCs作用 2 4h后 ,其G1期细胞数显著高于对照组 (94.65 %和 79.97% ) (P <0 .0 1) ,以H2 O2浓度在 10 0 μmol/L时尤为明显 ;流式细胞仪AnnexinV PI双标细胞凋亡检测 ,10 0 μmol/LH2 O2 有促进VSMCs凋亡作用 ,并在作用 48h左右达到高峰 ,占 2 6.76% ,与对照组相比 ,差异有显著性 (P <0 .0 1)。结论 H2 O2 浓度≥ 10 0 μmol/L时 ,可显著抑制VSMCs增殖 ;当H2 O2 浓度为 10 0 μmol/L时 ,有促进VSMCs凋亡作用。

ObjectiveTo study the mechanism of hydrogen peroxide (H_2O_2) to rabbit vascular smooth muscle cells.MethodsRabbit vascular smooth muscle cells (VSMCs) were primarily cultured,and MTT assay was performed at different durations to detect the proliferative effect of H_2O_2 on VSMCs and to determine the concentration of H_2O_2 for further study.Cell cycles and apoptosis of VSMCs after being exposed to certain concentration of H_2O_2 were observed by flow cytometry.ResultsIn MTT assay,when H_2O_2 ≥100 μmol/L,only 1-h exposure could cause a significant decrease of A values ( P < 0.01).In cell cycles detection,when H_2O_2 ≥100 μmol/L,after 24 h exposure,the percentage of VSMCs in stage G_1 was significantly increased as compared with controls (94.65% vs.79.97%, P < 0.01),especially in group of 100 μmol/L H_2O_2.In Annexin V and PI double stain for apoptosis detection,as compared with controls,100 μmol/L H_2O_2 could produce time-dependent apoptosis of VSMCs ( P < 0.01),and reached its peak value of 26.76% at 48 h.ConclusionH_2O_2 can significantly inhibit VSMCs proliferation while H_2O_2 ≥100 μmol/L.100 μmol/L of H_2O_2 had the effect of inducing apoptosis of VSMCs.