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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 observe the effect of crude extract of Blattella germanica (GCE) on the expression of PAR-2 and the secretion of vascular endothelial growth factor (VEGF) in human bronchial epithelial cells (HBE-16). Methods HBE-16 cells were randomly divided into normal control group and experimental group. The experimental group of cells were exposed to three different concentrations (25, 50 and 75μg / ml) of GCE, with or without TNF-α (3ng / ml) 24h. At the same time, 3 different concentrations of GCE + TNF-α group PAR-2 agonist SLIGKV-NH2. The expression of PAR-2 in HBE-16 cells was detected by RT-PCR and immunofluorescence. The level of VEGF in supernatant of each group was detected by ELISA. Results With the increase of GCE concentration, HBE-16PAR-2 expression was increased in a concentration-dependent manner (P <0.05). The level of VEGF in GCE + TNF-α group was higher than that in GCE group (P <0.01), GCE + TNF-α + SLIGKV group was higher than GCE + TNF-α group (P <0.05). Conclusion GCE up-regulates PAR-2 expression in HBE-16 cells in a concentration-dependent manner. TNF-α can cooperate with GCE to stimulate HBE-16 cells to secrete VEGF.