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目的:探讨法舒地尔对血小板源性生长因子(platelet-derived growth factor,PDGF)诱导的人肺动脉平滑肌细胞(human pulmonary artery smooth muscle cell,HPASMC)增殖的抑制作用及机制。方法:以体外培养的HPASMC为研究对象,PDGF-BB诱导增殖,法舒地尔进行预处理,MTT法检测细胞增殖;流式细胞仪检测细胞周期;Western blot检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和p27kip1蛋白的表达。结果:细胞培养24 h后与空白组比较,PDGF组HPASMC细胞增殖比例,S期细胞比例和PCNA蛋白表达明显增加,p27kip1蛋白表达则明显降低;用法舒地尔预处理后,与PDGF组比较,细胞增殖比例,S期细胞比例和PCNA蛋白表达明显下降,p27kip1蛋白表达则明显增加。结论:法舒地尔可通过上调p27kip1蛋白表达来抑制PDGF诱导的HPASMC增殖和细胞周期进程。
AIM: To investigate the inhibitory effect of fasudil on the proliferation of human pulmonary artery smooth muscle cell (HPASMC) induced by platelet-derived growth factor (PDGF) and its mechanism. Methods: HPASMC cultured in vitro was treated with PDGF-BB and pre-treated with Fasudil. The cell proliferation was measured by MTT assay. The cell cycle was detected by flow cytometry. The proliferating cell nuclear antigen , PCNA) and p27kip1 protein expression. Results: Compared with the blank group, the cell proliferation ratio, the proportion of S phase cells and the expression of PCNA protein in PDGF group were significantly increased and the expression of p27kip1 protein was significantly decreased in 24 h after cell culture. Compared with PDGF group, Cell proliferation ratio, S phase cell ratio and PCNA protein expression decreased significantly, p27kip1 protein expression was significantly increased. Conclusion: Fasudil inhibits PDGF-induced HPASMC proliferation and cell cycle progression by up-regulating p27kip1 protein expression.