摘要
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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2004年第4期403-405,共3页
Chinese Journal of Experimental Surgery
基金
上海市高等学校青年科学基金 (0 2BQ2 0 )
上海市卫生系统百人计划 (98BR0 0 6)
