尿激酶对大鼠胸膜成纤维细胞分泌TGF-β_1、VEGF、PAI-1的影响

The influence of urokinase on pleural fibroblasts secreting transforming growth factor β_1,vascular endothelial growth factor and plasminogen activator inhibitor-1 in rats

目的探讨尿激酶(urokinase,UK)对大鼠胸膜的成纤维细胞(fibroblasts,FB)分泌转化生长因子-β1(transforming growth factor beta-1,TGF-β1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、纤溶酶原激活物抑制剂(plasminogen activator inhibitor-1,PAI-1)的影响。方法取健康雄性SD大鼠胸膜,培养、纯化并鉴定FB。设对照组和实验组。对照组分2组:对照0组由不含胸膜FB的40个样本组成,加入无血清RPMI 1640培养液;对照1组由40个胸膜FB样本组成,加入无血清RPMI 1640培养液,培养24h后,用酶联免疫吸附法测定上清液中TGF-β1、PAI-1、VEGF的含量;实验组分为4组,共40个样本,分别加入5000、10000、20000和30000I U/ml的UK,培养24h后取上清液,测定其中TGF-β1、VEGF、PAI-1量。结果对照0组未检测出TGF-β1、PAI-1、VEGF;对照组1胸膜FB分泌TGF-β1、VEGF、PAI-1的量为(3 135.205±390.975)pg/mL;(22.09±7.48)ng/ml;(1.8775±0.39)ng/ml;加入5000～30000I U/ml尿激酶的试验组,TGF-β1、VEGF、PAI-1的浓度与对照1组相比,有明显下降(P<0.05)。结论胸膜成纤维细胞能分泌TGF-β1、PAI-1、VEGF;而尿激酶能抑制其分泌。

Objective To study the secretion of transforming growth factorβ1 （TGF-β1）, vascular endothelial growth factor （VEGF） and plasminogen activator inhibitor-1 （PAI-1）by fibroblasts （FB） and the influence of urokinase（UK） on the secretion. Methods Fibroblasts obtained from the pleura of fine male rats were cultured, purified and identified, and then were divided into control group 0, control group 1 and experimental group. There were 40 samples without fibroblasts in RPMI 1640 medium in control group 0 and 40 samples with many fibroblasts in RPMI 1640 medium in control group 1. 5 000,10 000,20 000 and 30000 IU/ml of urokinase were added into the 40 samples in four experimental groups, respectively. The samples each group were cultured for 24 hours, and then the level of TGF-β1, VEGF and PAI-1 in cultured supernatant fluids were measured by enzyme linked immunosorbent assay（ELISA）. Results TGF-β1, VEGF and PAI-1 were not detected in control group 0. The concentration of TGF-β1 ,VEGF and PAl-1 in control group 1 were （3135. 205 ±390. 975）pg/ml, （22.09±7.48）ng/ml, （1.8775±0.39） ng/ml, respectively. They decreased obviously in four experimental groups compared with control groups. Conclusions Pleural Fibroblasts could secrete TGF-β1, VEGF and PAI-1. Urokinase could inhibit their secretion.