尿激酶型纤溶酶原激活物对高糖诱导大鼠系膜细胞增殖及表型转化的影响及其机制  

Effects of urokinase-type plasminogen activator on high glucose-induced rat mesangial cellsproliferation and phenotype transformation

作　　者：程晖 丁国华 陈铖 石明 杨红霞 

机构地区：[1]大学人民医院肾内科,武汉430060

出　　处：《中华肾脏病杂志》2011年第7期515-519,共5页Chinese Journal of Nephrology

摘　　要：目的通过体外培养大鼠系膜细胞（MC），观察尿激酶型纤溶酶原激活物（uPA）对高糖环境下MC增殖及表型转化的影响及其可能的信号转导机制。方法体外培养大鼠MC，分为4组：对照组、高糖组、高糖＋渥曼青霉素组、高糖＋uPA组。MTF法检测各组MC增殖情况。流式细胞仪分析各组MC细胞周期改变。Western印迹法检测各组ME表达CDK2与p27^kipl变化，并测定MC中信号蛋白Akt的活性。激光共聚焦显微镜检测各组MC中α-SMA表达方式及表达量变化。结果培养24h后高糖组MC增殖程度较对照组显著增加（P〈0．01），渥曼青霉素与uPA组可明显抑制细胞增殖（P〈0．01）。高糖刺激Akt活性，渥曼青霉素与uPA组Akt活性均较高糖组显著降低（均P〈0．01）。高糖组MC培养24h后，p27^kipl蛋白表达较对照组显著减少（P〈0．01）；高糖＋渥曼青霉素组、高糖＋uPA组p27^kipl蛋白表达均较高糖组显著增加（均P〈0．01）；各组CDK2蛋白表达无明显变化。高糖组MC培养24h后，胞质中仪．SMA表达较对照组显著增加（P〈O．01），并在核周出现聚集；高糖＋渥曼青霉素组、高糖＋uPA组α—SMA表达量均较高糖组显著减少（均P〈0．01），其分布与对照组无显著差异。结论uPA可能通过抑制Akt信号分子活性，上调p27^kipl表达，拮抗高糖所致MC增殖与表型转化。Objective To explore the effects and mechanisms of urokinase-type plasminogen activator （uPA） on high glucose-induced rat mesangial cells proliferation and phenotype transformation. Methods Rat mesangial cells were cultured and incubated in media containing either 5 mmol/L D-glucose or 30 mmol/L D-glucose with or without addition of wortmannin, or uPA （105 U/L） for different time periods. At the end of the incubation period, mesangial cells proliferation was assessed by MTT assay and flow cytometric analysis. Cyelin-dependent kinase 2 （CDK2） and p27^kipl expression and activation of Akt were evaluated by Western blotting and Akt kinase assay respectively. Furthermore, the expression and distribution of α-SMA were detected with laser confoeal microscopy. Results MTT assay and flow eytometrie analysis demonstrated that high glucose induced mesangial cells proliferation （P〈0.05） and an incresed proportion of cells in G2/M＋S stage after 24 h incubation （P〈0.01）, which were attenuated by uPA or wortmannin （P〈0.01）. High glucose induced the enhance of Akt activity after 3 h （P〈0.05）, and the effect was inhibited by wortmannin or uPA （P〈0.01）. High glucose did not alter CDK2 expression （P〉0.05）, but significantly inhibited p27^kipl expression （P〈0.05）, which was attenuated by wortmannin or uPA （P〈0.01）. High glucose induced the up-regulation of α-SMA expression and perinucleus location in mesangial cells after 24 h （P〈0.01）, which were alleviated by wortmannin or uPA （P〈0.01）. Conclusion uPA up-regulates p27^kipl expression and counteracts high glucose-induced mesangial cells proliferation and phenotype transformation via blocking PI3K-Akt signaling pathway.

关 键 词：尿激酶型纤溶酶原激活物 系膜细胞 增殖 信号转导 

分 类 号：R363[医药卫生—病理学]