南方医科大学学报

南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (10): 1322-.

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乌司他丁通过上调PPAR-γ抑制低氧诱导的肺血管平滑肌细胞表型转换

唐锟，刘畅，陈林，高静，张超   

  • 出版日期:2016-10-20 发布日期:2016-10-20

Ulinastatin inhibits hypoxia-induced phenotype modulation of pulmonary artery smooth muscle cells by up-regulating PPAR-γ expression

  • Online:2016-10-20 Published:2016-10-20

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摘要： 目的探讨乌司他丁对低氧诱导的肺血管平滑肌细胞（PASMCs）表型转化的影响及其分子机制。方法体外培养SD大鼠 PASMCs，随机分组4组：正常组（N组），常氧+乌司他丁组（NU组），低氧组（H组），低氧+乌司他丁组（HU组），各组培养24 h后 采用免疫荧光检测SM-α-actin和Calponin表达水平，应用CCK-8法、3H-TdR和Transwell小室检测各组PASMCs增殖和迁移。 并分别利用Western blotting和荧光素酶报告质粒检测PPAR-γ蛋白表达和转录活性。采用PPAR-γ抑制剂GW9662进行预处理 干预乌司他丁对低氧诱导的PASMCs表型转化的影响，分析干预后PASMCs增殖和迁移改变。结果低氧条件下，乌司他丁能 促进PASMCs 中SM-α-actin 和Calponin 的表达（P<0.05），抑制PASMCs 的增殖和迁移能力（P<0.05），同时乌司他丁上调 PPAR-γ的表达（P<0.05）。采用GW9662预处理后，乌司他丁对低氧诱导的PASMCs表型转换的抑制作用显著下降（P<0.05）。 结论乌司他丁通过上调PPAR-γ抑制低氧诱导的PASMCs表型转换。

Abstract: Objective To evaluate the effect of ulinastatin on hypoxia-induced phenotype modulation of pulmonary artery smooth muscle cells (PASMCs) and explore the underlying mechanism. Method Cultured PASMCs from SD rats were exposed to normoxic condition, normoxia with ulinastatin treatment, hypoxia, or hypoxia with ulinastatin treatment. After 24 h of exposures, the cells were examined for SM-α-actin and caplonin expressions with immunofluorescence assay and for cell migration with CCK-8 and 3H-TdR assays. Western blotting was used for detecting the expressions of PPAR-γ in the cells, and PPAR-γ-responsive firefly luciferase reporter was employed for measuring the transcriptional activity of PPAR-γ. The PPAR-γ inhibitor GW9662 was used to explore the mechanism of the inhibitory effect of ulinastatin on hypoxia induced-phenotype modulation of PASMCs by measuring the changes in cell proliferation and migration. Results Ulinastatin obviously enhanced the expressions of SM-α-actin and calponin (P<0.05), inhibited the proliferation and migration (P<0.05), and up-regulated the expression of PPAR-γ in PASMCs exposed to hypoxia (P<0.05). Pretreatment of the cells with GW9662 abolished the effect of ulinastatin on hypoxia-induced phenotype modulation of PASMCs and enhanced the cell proliferation and migration (P<0.05). Conclusion Ulinastatin inhibits hypoxia-induced phenotype modulation of PASMCs from rats possibly by up-regulating the expression of PPAR-γ.