IFN-γ诱导的肾小管上皮细胞CXCL9、CXCL10和CXCL11的表达  

Expressions of CXCL9,CXCL10 and CXCL11 in renal tubular epithelial cells induced by IFN-γ

作　　者：林青[1] 宋艳芳[1] 祝先进[2] 杨顺良[3] 郑健[4] 

机构地区：[1]福建中医药大学附属人民医院检验科,福建福州350004 [2]福建医科大学附属协和医院检验科,福建福州350001 [3]南京军区福州总医院泌尿外科,福建福州350025 [4]福建中医药大学,福建福州350001

出　　处：《细胞与分子免疫学杂志》2013年第2期137-140,145,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：福建省医学创新课题(2011-CX-28);福建省教育厅课题(JB11066);福建中医药大学校管课题(XB2011020)

摘　　要：目的探讨IFN-γ对人近端肾小管上皮细胞(HK-2)趋化因子CXCL9、CXCL10和CXCL11分泌的影响以及趋化因子的功能。方法 IFN-γ分别作用HK-2细胞不同时间后,用real-time PCR检测其CXCL9、CXCL10和CXCL11 mRNA的表达,以ELISA检测细胞培养上清中分泌趋化因子CXCL9、CXCL10和CXCL11的蛋白水平,用流式细胞术检测淋巴细胞表面CXCR3表达情况,通过趋化实验检测IFN-γ作用HK-2细胞培养上清对淋巴细胞的趋化作用。结果 IFN-γ作用12 h后,HK-2细胞分泌趋化因子开始升高,CXCL9 mRNA的表达在48 h达到高峰,CXCL10、CXCL11 mRNA的表达在24 h达到高峰。在蛋白水平上,HK-2细胞经IFN-γ诱导12 h后,开始分泌趋化因子蛋白,CXCL9、CXCL11的分泌在IFN-γ作用HK-2细胞72 h达到高峰,CXCL10的分泌在48 h达到高峰。与新鲜分离的淋巴细胞相比活化的淋巴细胞表面的CXCR3表达明显升高。IFN-γ作用HK-2细胞培养上清对活化的淋巴细胞具有明显的趋化作用且能被抗CXCR3抗体所阻断。结论 IFN-γ能够明显上调肾小管上皮细胞趋化因子CXCL9、CXCL10和CXCL11 mRNA表达和蛋白的分泌。Objective To investigate the effect of interferon-γ（IFN-γ） on the releases of CXCL9,CXCL10 and CXCL11 in renal proximal tubular epithelial cells（HK-2） and explore their functions.Methods After stimulated by IFN-γ for different time,the HK-2 cells were analyzed by real-time PCR to detect the expressions of CXCL9,CXCL10 and CXCL11 mRNA,and were tested by ELISA to quantify the releases of CXCL9,CXCL10 and CXCL11 in supernatants.After activation of lymphocytes,cells were examined by flow cytometry to determine the CXCR3 expression,while chemotaxis assay was applied to evaluate the chemotactic effect of the supernatants of IFN-stimulated HK-2 cells on lymphocytes.Results Compared with the control group,the expressions of CXCL9,CXCL10 and CXCL11 mRNA in the IFN-stimulated group were significantly increased 12 h after stimulation,and peaked at 48,24 and 24 h,respectively.The levels of CXCL9,CXCL10 and CXCL11 proteins began to rise at 12 h and reached the peak at 72,48 and 72 h,respectively.Compared with the control group,the expression of CXCR3 was markedly increased in the activated lymphocytes.The activated lymphocytes were recruited by the culture supernatants of IFN-stimulated HK-2 cells and the pretreatment of CXCR3 antibody inhibited this chemotactic effect.Conclusion IFN-γ can significantly up-regulate the expressions of chemokines such as CXCL9,CXCL10 and CXCL11 at both mRNA and protein levels in HK-2 cells.

关 键 词：干扰素-Γ CXCL9 CXCL10 CXCL11 人肾小管上皮细胞 

分 类 号：R737.31[医药卫生—肿瘤]