Wnt-5a/JNK信号通路参与PM2.5诱导的人支气管上皮细胞系16HBE分泌IL-6和TNF-α

Wnt-5a/JNK is involved in PM2.5-induced expression of IL-6 and TNF-α of human bronchial epithelial cell line 16HBE

目的探讨PM2.5是否通过Wnt-5a/JNK信号通路诱导人支气管上皮细胞系16HBE中IL-6和TNF-α的分泌。方法本研究分为大鼠和16HBE细胞两部分。具体如下:将大鼠分为对照组和吸入机动车尾气组(浓度1.5 mg/m2,2 h/d,1月);将16HBE细胞分为对照组、PM2.5组(20μg/mL)、阴性对照siRNA+PM2.5组、Wnt-5a siRNA+PM2.5组、Wnt-5a siRNA+对照组、JNK抑制剂SP600125+PM2.5组、SP600125+对照组。采用免疫组化检测肺组织p-JNK和p-c-Jun表达水平;Western blot检测细胞中Wnt-5a、JNK、c-Jun、p-JNK和p-c-Jun表达水平;ELISA检测IL-6和TNF-α的分泌。结果与大鼠对照组相比,机动车尾气组的p-JNK和p-c-Jun表达水平显著增加(P<0.05);与16HBE细胞对照组相比,PM2.5组的Wnt-5a、p-JNK、p-c-Jun、IL-6和TNF-α的表达水平显著增加(P<0.05);与PM2.5组相比,Wnt-5a siRNA+PM2.5组Wnt-5a、p-JNK、p-c-Jun、IL-6和TNF-α的表达水平显著减少(P<0.05),SP600125+PM2.5组p-c-Jun、IL-6和TNF-α的表达水平显著减少(P<0.05)。结论Wnt-5a/JNK信号通路参与了PM2.5诱导的人支气管上皮细胞系分泌IL-6和TNF-α。

Objective To explore whether PM2.5 induces the secretion of IL-6 and TNF-αin 16HBE via the Wnt-5a/JNK signaling pathway.Methods The research was implemented by rats and 16HBE cells.Rats were divided into control group and motor vehicle exhaust(MVE)group(concentration 1.5 mg/m2,2 h/day);16HBE cells were divided into control group,PM2.5(20μg/mL)group,control siRNA+PM2.5 group,Wnt-5a siRNA+PM2.5 group,Wnt-5a siRNA+control group,JNK inhibitor SP600125+PM2.5 group and SP600125+control group.The expressions of p-JNK and p-c-Jun in lung tissues were examined by immune-histochemistry method.The expressions of Wnt-5a,JNK,c-Jun,p-JNK and p-c-Jun were measured by Western blot.The secretions of IL-6 and TNF-αwere assessed by ELISA.Results Compared with the control rat group,MVE group exhibited significantly up-regulated the expression of p-JNK and p-c-Jun(P<0.05);Compared with the 16HBE cell control group,PM2.5 group significantly up-regulated the expressions of Wnt-5a,p-JNK and p-c-Jun and the secretions of IL-6 and TNF-α(P<0.05);Compared with the PM2.5 group,Wnt-5a siRNA+PM2.5 group blocked these effects(P<0.05).Furthermore,SP600125+PM2.5 group showed a significantly decreased expression of p-c-Jun and the secretion of IL-6 and TNF-αby 16HBE cells(P<0.05).Conclusions PM2.5 induces the secretions of IL-6 and TNF-αby 16HBE cells via Wnt-5a/JNK signaling pathway.