【摘要】 目的探讨神经生长因子(NGF)通过c-fos影响支气管上皮细胞NK-1R的表达。方法以人支气管上皮细胞(NHBEC)为研究对象,通过阳离子脂质体将c-fos siRNA转染至细胞内,然后用NGF干预。实验设五组:对照组、NGF干预组、NGF+c-fos siRNA组、NGF+空白脂质体组、NGF+非特异性dsRNA组。转染12 h后,用NGF干预60 min,采用Western blot法测定c-fos蛋白表达,采用免疫细胞化学法和RT-PCR从蛋白和mRNA水平观察细胞中NK-1R的表达。结果与对照组比,NGF组c-fos蛋白、NK-1R蛋白和NK-1RmRNA表达均明显增加(P<0.01);与NGF组比,NGF+c-fos siRNA组中c-fos蛋白、NK-1R蛋白和NK-1RmRNA表达均明显减少(P<0.01),而NGF组、NGF+空白脂质体组、NGF+非特异性dsRNA组之间的c-fos蛋白、NK-1R蛋白和NK-1RmRNA表达无显著差异。结论 NGF可通过c-fos促进支气管上皮细胞中NK-1R的表达,介导哮喘神经源性炎症,RNAi技术可以减少NGF诱导的人支气管上皮细胞c-fos蛋白及NK-1R的表达。更多还原

【Abstract】 Objective To investigate whether the effect of nerve growth factor on NK-1R expression of bronchial epithelial cells is via c-fos. Methods Human bronchial epithelial cells( NHBEC) were taken as the research object,the c-fos siRNA was transected into cells through the positive ion fat plastid,and then the NGF intervention was carried on. The control group,NGF intervention group,NGF + c-fos siRNA group,NGF +liposomes group,NGF + non-specific dsRNA group were decided. After trans-infection for 12 h and then intervention for 60 min,the protein expression of c-fos was detected by western-blot. The protein and mRNA expression of NK-1R was determined by RT-PCR and immune cell chemical definite quality respectively.Results Compared with the control group,the expressions of c-fos protein,NK-1R protein and NK-1RmRNA were significantly increased in NGF group( P < 0. 01); compared with NGF group,the expressions of c-fos protein,NK-1R protein and NK-1RmRNA were significantly reduced in NGF+c-fos siRNA group( P< 0.01),NGF intervention group,NGF+liposomes group,NGF+ non-specific dsRNA group had no significant difference in the expression of them. Conclusion NGF stimulates the expressions of NK-1R in bronchial epithelial cells via c-fos. and then mediate neurogenic inflammation of asthma. RNAi could reduced the of c-fos protein and NK-1R induced by NGF in human bronchial epithelial cells.更多还原