Lin Qing, Song Yanfang, Zhu Xianjin, Yang Shunliang, Zheng Jian
Department of Clinical Laboratory, Fujian University of Traditional Chinese Medicine, Fuzhou, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Feb;29(2):137-40.
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.
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.
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.
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.
探讨干扰素-γ(IFN-γ)对肾近端小管上皮细胞(HK-2)中CXCL9、CXCL10和CXCL11释放的影响,并探究其功能。
用IFN-γ刺激不同时间后,采用实时PCR分析HK-2细胞中CXCL9、CXCL10和CXCL11 mRNA的表达,并用ELISA检测上清液中CXCL9、CXCL10和CXCL11的释放量。淋巴细胞活化后,通过流式细胞术检测CXCR3的表达,同时采用趋化试验评估IFN刺激的HK-2细胞上清液对淋巴细胞的趋化作用。
与对照组相比,IFN刺激组中CXCL9、CXCL10和CXCL11 mRNA的表达在刺激后12 h显著增加,分别在48、24和24 h达到峰值。CXCL9、CXCL10和CXCL11蛋白水平在12 h开始升高,分别在72、48和72 h达到峰值。与对照组相比,活化淋巴细胞中CXCR3的表达明显增加。活化淋巴细胞被IFN刺激的HK-2细胞培养上清液募集,CXCR3抗体预处理可抑制这种趋化作用。
IFN-γ可显著上调HK-2细胞中CXCL9、CXCL10和CXCL11等趋化因子在mRNA和蛋白水平的表达。