摘要
目的探讨乙型肝炎病毒x基因（HBX）转染人近端肾小管上皮细胞（HK-2）后对其凋亡及免疫指标的影响。方法体外培养HK-2。用分子克隆的方法构建pcDNA3．1-myc-HBX质粒，采用脂质体转染法瞬时转染HK-2细胞。实时定量PCR和Western印迹法检测细胞中Toll样受体4（TLR4）的变化，流式细胞术检测细胞凋亡情况及细胞表面人组织相容复合物（MHC）-Ⅱ、CD40的变化；ELISA法检测细胞培养上清中IL-4、IFN-γ水平。结果与对照组相比，转染pcDNA3．1-myc-HBX质粒组凋亡细胞数量显著增多（P〈0．05）；TLR4的表达及其表面MHC-Ⅱ、CD40表达均显著上调（均P〈0．05）；其上清液中IFN-γ水平增加（P〈0．05），但IL-4水平降低（P〈0．05）。结论HBX质粒转染HK-2细胞后可引起细胞凋亡增加，促进细胞表面免疫分子的表达，最终导致细胞损伤及免疫微环境紊乱。
Objective To investigate the effect of hepatitis B virus X （HBX） gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line （HK- 2）. Methods The eukaryotic vector peDNA3.1- mye- HBX containing HBX gone was transiently transfected into HK-2 cells by lipofeetamine mediation. Untransfeeted HK-2 cells and those transfected with empty vector were used as controls. The TLR4 expression was detected by real-time PCR and Western blotting. The apoptosis of cells and expression of MHC- Ⅱ and CD40 were detected by flow cytometry, and the contents of IL-4 and IFN-γ in the supernatant were detected by ELISA. Results Compared with control groups, the number of apoptotic cells was significantly increased in the HBX transfection group （P 〈 0.05）, and the expressions of TLR4, MHC- Ⅱand CD40 were also significantly increased in the HBX transfection group （all P〈0.05）. IFN-γ level in the supernatant of HBX transfection group was higher （P 〈 0.05）, but IL-4 level was lower as compared to control groups （all P 〈 0.05）. Conclusions Over-expression of HBX gene may induce apoptosis of HK-2 cells and up- regulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2013年第1期50-54,共5页
Chinese Journal of Nephrology
基金
国家自然科学基金（81170672）
