德国小蠊粗提物对人支气管上皮细胞蛋白酶活化受体2表达及血管内皮生长因子分泌的影响

Effects of German cockroach extract on expression of PAR-2 and secresion of VEGF from HBE-16

目的观察德国小蠊粗提物(GCE)对人支气管上皮(HBE-16)细胞蛋白酶活化受体2(PAR-2)表达及血管内皮生长因子(VEGF)分泌的影响。方法将HBE-16细胞随机分为正常对照组和实验组。实验组细胞分别暴露于3个不同浓度(25、50和75μg/ml)的GCE,每组加或不加TNF-α(3ng/ml)24h。同时在3个不同浓度GCE+TNF-α组中加入PAR-2的激动剂SLIGKV-NH2。RT-PCR、免疫荧光法检测HBE-16细胞PAR-2表达。ELISA检测各组细胞上清液VEGF水平。结果随着GCE浓度的增加,HBE-16PAR-2的表达呈浓度依赖性上调(P<0.05)。GCE+TNF-α组较GCE组细胞上清液VEGF水平增加(P<0.01),GCE+TNF-α+SLIGKV组较GCE+TNF-α组增加(P<0.05)。结论 GCE呈浓度依赖性上调HBE-16细胞PAR-2表达。TNF-α可协同GCE刺激HBE-16细胞分泌VEGF。

Objective To investigate the effects of German cockroach extract（GCE）on the expression of protease-activated receptor 2（PAR-2）and secresion of vascular endothelial growth factor（VEGF）from human bronchial epithelium（HBE）-16 cells.Methods HBE-16 cells in group T were routinely cultured and divided into two groups of C（control cells）and T（treated group）.HBE-16 cells in group T were treated with different concentrations（25,50 and 75μg/ml）of GCE,in which SLIGKV-NH2（an agonist of PAR-2）with or without TNF-α was added.RT-PCR and immunofluresence were used to detect the expression of PAR-2in HBE-16 cells.VEGF level in supernatant was detected by ELISA.Results The expression of PAR-2 was increased in a concentration-dependent manner with the increase of GCE（P〈0.05）.VEGF levels in supernatant were higher in groups of GCE＋TNF-αthan those in groups of GCE（P〈0.01）,which were higher in groups of GCE＋TNF-α＋SLIGKV-NH2 than those in groups of GCE＋TNF-α（P〈0.05）.Conclusion GCE may upregulate the expression of PAR-2in a concentration-dependent manner in HBE-16 cells.TNF-αmay enhance the secresion of VEGF in HBE-16 cells stimulated by GCE.