Wu Sheng-hua, Lu Chao, Dong Ling, Zhou Guo-ping
Department of Pediatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Jan;22(1):37-9.
To examine whether connective tissue growth factor(CTGF) induces the production of fractalkine(FLK) by glomerular mesangial cells of rats, and explore the mechanism of signal pathway of CTGF actions.
The mRNA expression of FLK was analyzed by RT-PCR in cultured mesangial cells stimulated by CTGF. The protein of FLK in the supernatants of cells was determined by ELISA. The chemotactic effect of the supernatants on monocytes was assessed by the in vitro chemotaxis assay. The phosphorylation of p42/44 MAPK was assessed by Western blot.
Treatment of the cells with CTGF enhanced the mRNA expression of FLK and concentration of FLK in the supernatants. Pretreatment of the supernatants of CTGF-treated cells with anti-FLK antibodies partially inhibited the chemotactic effect of the supernatants on monocytes. CTGF increased the p42/44MAPK phosphorylation. Pretreatment of the cells with PD98059 or UO126, inhibitors of phosphorylated p42/44 MAKP, decreased the CTGF-induced expression of phosphorylated p42/44 MAPK and concentration of FLK in supernatants.
CTGF-induced secretion of FLK by mesangial cells was associated with the phosphorylation of p24/p44 MAPK.
研究结缔组织生长因子（CTGF）是否能诱导大鼠肾小球系膜细胞产生趋化因子（FLK），并探讨CTGF作用的信号通路机制。
采用逆转录聚合酶链反应（RT-PCR）分析CTGF刺激培养的系膜细胞中FLK的mRNA表达。用酶联免疫吸附测定（ELISA）法测定细胞上清液中FLK的蛋白含量。通过体外趋化试验评估细胞上清液对单核细胞的趋化作用。用蛋白质免疫印迹法评估p42/44丝裂原活化蛋白激酶（MAPK）的磷酸化。
用CTGF处理细胞可增强FLK的mRNA表达及细胞上清液中FLK的浓度。用抗FLK抗体预处理CTGF处理细胞的上清液可部分抑制上清液对单核细胞的趋化作用。CTGF可增加p42/44 MAPK的磷酸化。用磷酸化p42/44 MAPK的抑制剂PD98059或UO126预处理细胞，可降低CTGF诱导的磷酸化p42/44 MAPK的表达及细胞上清液中FLK的浓度。
CTGF诱导系膜细胞分泌FLK与p24/p44 MAPK的磷酸化有关。
