内皮素-1促进人肺血管平滑肌细胞中细胞周期依赖激酶2的表达

The study of endothelin-1 prompting cycle dependent kinase 2 protein expression in human pulmonary artery smooth muscle cells

目的探讨内皮素-1对人肺血管平滑肌细胞(HPASMCs)增殖过程中细胞周期依赖激酶2(CDK2)的影响。方法取材肺癌周围的正常组织,将培养的HPASMCs统一到G0期接种,随机分为4组(每组4个样本):对照组不加任何干预因素,ET-1组10-7mol/L;ET-1拮抗剂BQ123组10-6mol/L;混合组ET-1 10-7mol/L+BQ123 10-6mol/L。采用免疫印迹方法,流式细胞仪分别检测了CDK2蛋白质的表达、细胞周期时相的变化。结果W estern blot分析结果表明:对照组,混合组CDK2蛋白质表达基本相同;ET-1组表达最高,与对照组比较差异有统计学意义(P<0.01);BQ123组CDK2蛋白质表达降低,但与对照组比较无明显差异,ET-1组与BQ123组比较CDK2蛋白质表达差异有统计学意义(P<0.01);流式细胞仪测定结果显示:与对照组比较,ET-1组表现为HPASMCs中G1相的比例减少,而S相增高(P<0.01);BQ123的G1相变化略有增高、S相变化略有减少;对照组,混合组变化基本相同,混合组、BQ123组与对照组比较差异无统计学意义(P>0.05);ET-1组与BQ123组各个时相比较差异有统计学意义(P<0.01)。结论内皮素-1促进人肺血管平滑肌细胞CDK2的表达,在肺血管平滑肌细胞增殖方面可能有着重要的病理生理学意义。

Objective To explore the relationship between endothelin-1 （ET-1） and cycle dependent kinase 2 （CDK2） in human pulmonary artery smooth muscle ceils （HPASMCs） proliferation. Method Cultured HPASMCs were divided into four groups： control group, ET-1 group, BQ123 group and mixture group. The cultured HPASMCs were treated with 10^-7 mol/L ET-1 in ET-1 group, with 10^-6 mol/L BQ123 in BQ123 group, and with 10^-7 mol/L ET-1 plus 10^-6 mol/L BQ123 in mixture group, respetively. The expression of CDK2 protein was detected by Western blot and the cell cycle was analyzed by flow cytometry. Result The results of western blot showed that there was no significant difference in the CDK2 protein expression between the control group and mixture group. The expression of CDK2 protein in ET-1 group was the highest among the four groups （P 〈 0.01）. The expression of CDK2 protein was decreased in BQ123 group and there was no significant difference in the expression of CDK2 between the BQ123 group and the control group （P 〉 0.05）. The proportion of cells at G1 phase was decreased significantly while the cells at S phase was increased significantly in ET-1 group with the analysis of cell cycle by flow cytometry. There was no significant difference in the change of phase among the BQ123 group, mixture group and control group （P 〉 0.05）. Conclusion The ET-1 promotes the expression of CDK2 protein in HPASMCs, Which may play an important role in the proliferation of PASMCs.