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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 effects of IFN-γ on the secretion of chemokines CXCL9, CXCL10 and CXCL11 in proximal human proximal tubular epithelial cells (HK-2) and the function of chemokines. Methods The expression of CXCL9, CXCL10 and CXCL11 mRNA was detected by real-time PCR after treated with IFN-γ for different time. The levels of CXCL9, CXCL10 and CXCL11 in the cell culture supernatant were detected by ELISA . The expression of CXCR3 on lymphocytes was detected by flow cytometry. The chemotactic effect of IFN-γ on the lymphocytes was detected by chemotaxis assay. Results After induced by IFN-γ for 12 h, the chemotactic factors of HK-2 cells began to increase. The expression of CXCL9 mRNA peaked at 48 h. The expression of CXCL10 and CXCL11 mRNA peaked at 24 h. At the protein level, HK-2 cells started to secrete chemokine proteins after induced by IFN-γ for 12 h. The secretion of CXCL9 and CXCL11 peaked at 72 h in IFN-γ-treated HK-2 cells. CXCL10 secreted at 48 h Peak. CXCR3 expression on the surface of activated lymphocytes is significantly increased compared to freshly isolated lymphocytes. Effect of IFN-γ HK-2 cell culture supernatants have obvious chemotactic effect on activated lymphocytes and can be blocked by anti-CXCR3 antibody. Conclusion IFN-γ can upregulate the expression of CXCL9, CXCL10 and CXCL11 and the secretion of protein in renal tubular epithelial cells.