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目的:研究热休克蛋白90(HSP90)对血小板衍生因子(Platelet derived growth factor,PDGF)诱导的大鼠主动脉平滑肌细胞增殖的影响。方法:采用胶原酶消化法原代培养大鼠胸主动脉平滑肌细胞,应用脂质体细胞转染siRNA的方法抑制HSP90的表达,定量PCR和western blot的方法检测抑制效率。利用PDGF-bb诱导刺激平滑肌细胞增殖,CCK8法检测细胞增殖能力的变化,流式细胞术检测细胞生长周期的改变。结果:平滑肌细胞中转染HSP90的siRNA后,HSP90的mRNA和蛋白水平明显降低,分别为对照组的65.3%和57.6%(P<0.05);PDGF-bb刺激明显促进平滑肌细胞生长,而降低HSP90水平显著影响PDGF-bb诱导的细胞增殖(P<0.05);流式细胞术检测发现降低HSP90水平引起平滑肌细胞生长停滞,分布在细胞周期G1期的细胞比例明显增多(P<0.05)。结论:HSP90通过调控平滑肌细胞的生长周期参与调节细胞增殖过程。
Objective: To investigate the effect of heat shock protein 90 (HSP90) on the proliferation of rat aortic smooth muscle cells induced by platelet derived growth factor (PDGF). Methods: Thoracic aorta smooth muscle cells were primarily cultured by collagenase digestion. The expression of HSP90 was inhibited by lipofection of siRNA. The inhibitory efficiency was detected by quantitative PCR and western blot. Proliferation of smooth muscle cells was stimulated by PDGF-bb, the proliferation of cells was detected by CCK8 assay, and the changes of cell cycle were detected by flow cytometry. Results: The mRNA and protein levels of HSP90 in smooth muscle cells transfected with HSP90 were significantly decreased (65.3% and 57.6%, respectively) (P <0.05). PDGF-bb stimulated the proliferation of smooth muscle cells and decreased the expression of HSP90 (P <0.05). The results of flow cytometry showed that the decrease of HSP90 caused the arrest of smooth muscle cell growth, and the proportion of cells distributed in G1 phase increased significantly (P <0.05). Conclusion: HSP90 is involved in the regulation of cell proliferation by regulating the growth cycle of smooth muscle cells.