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目的:探讨内皮素-1是否通过细胞周期蛋白质D1与细胞外调节蛋白激酶通路促进人脐动脉平滑肌细胞增殖。方法:采用MTT法观察ET-1和PD98059对人脐动脉平滑肌细胞生长的作用;[~3H]TdR法观察对细胞DNA合成的作用;流式细胞仪法观察对细胞增殖周期的影响;蛋白质印迹法观察对细胞外调节蛋白激酶和细胞周期蛋白质D1表达的影响。结果:首先,同没有ET-1组和PD98059组比较,ET-1促进平滑肌细胞增殖(P<0.05)。PD98059抑制ET-1诱导的血管平滑肌细胞增殖。第二,与没有ET-1组比较,ET-1促进平滑肌细胞DNA合成(P<0.05)。第三,ET-1促进平滑肌细胞增殖周期从G_0/G_1期向S期的转变,与没有ET-1组比较,G_0/G_1期细胞百分比明显减少,S期细胞百分比明显增加(P<0.05)。第四,ET-1增加细胞外信号调节性激酶的磷酸化水平和细胞周期蛋白质D1的蛋白表达,ERK的抑制剂可以抑制细胞外信号调节性激酶的磷酸化水平和细胞周期蛋白质D1的蛋白表达,与没有ET-1组比较,磷酸化-ERK和细胞周期蛋白质D1表达明显增强,对非磷酸化ERK表达没有影响。结论:内皮素-1可以通过细胞周期调节素D1与细胞外信号调节性激酶通路促进平滑肌细胞增殖。
AIM: To investigate whether endothelin-1 promotes the proliferation of human umbilical artery smooth muscle cells through cyclin D1 and extracellular regulated protein kinase pathways. Methods: The effects of ET-1 and PD98059 on the proliferation of human umbilical artery smooth muscle cells were observed by MTT assay. The effect of [~ 3H] TdR on DNA synthesis was observed. The effect of ET-1 and PD98059 on cell cycle was observed by flow cytometry. The effect on extracellular regulated protein kinase and cyclin D1 expression was observed. Results: Firstly, ET-1 promoted the proliferation of VSMCs (P <0.05) as compared with ET-1 group and PD98059 group. PD98059 inhibits ET-1-induced proliferation of vascular smooth muscle cells. Second, ET-1 promoted the DNA synthesis of smooth muscle cells compared with no ET-1 group (P <0.05). Third, ET-1 promoted the transition from G_0 / G1 phase to S phase in smooth muscle cells. Compared with ET-1 group, the percentage of cells in G_0 / G_1 phase decreased significantly and the percentage of cells in S phase increased significantly (P <0.05) . Fourth, ET-1 increased phosphorylation of extracellular signal-regulated kinase and protein expression of cyclin D1, and inhibitors of ERK inhibited phosphorylation of extracellular signal-regulated kinase and protein expression of cyclin D1 , Compared with no ET-1 group, phosphorylation-ERK and cyclin D1 expression was significantly enhanced, and had no effect on non-phosphorylated ERK expression. Conclusion: Endothelin-1 can promote the proliferation of smooth muscle cells through the interaction between cyclin D1 and extracellular signal-regulated kinase.