Effect of transforming growth factor-beta 1 on stromal cell-derived factor 1 expression in rat renal tubular epithelial cells / 中国组织工程研究

ABSTRACT

AIM: It has been proved that transforming growth factor-?1 (TGF-?1) has a close correlation with kidney fibrosis. This study investigated the effects of TGF-?1 on stromal cell-derived factor 1 (SDF-1) in rat renal tubular epithelial cells (NRK52E). METHODS: The experiment was carried out in the Division of Molecular Neurobiology, State Key Laboratory of Biotherapy, Sichuan University from March 2006 to May 2007. ①Rat renal tubular epithelial cells (NRK-52E, Monash Medical Center, Australia) were divided into normal control group, which were not treated with TGF-?1 (Cytola), and experimental group, which were subdivided into time-dependent groups NRK-52E cells were incubated with TGF-?1 at the same concentration (2 ?g/L) for 6, 12, and 24 hours, and dose-dependent groups NRK-52E cells were treated with TGF-?1 at different concentrations (2, 5, and 10 ?g/L) for 24 hours. ②The semi-quantitative calculation of SDF-1 protein expression in the cells treated with TGF-?1 at the concentration of 2 ?g/L for different time was accessed by immunocytochemistry to find the best time point. Then the variances of the mRNA and protein expression level in cells treated with TGF-?1 at different concentrations for 24 hours were detected by RT-PCR and Western-Blotting. RESULTS: ①The protein expression level of SDF-1 in NRK-52E cells treated with TGF-?1 for 12 and 24 hours was higher than for 0 hour (P 0.05). This indicated the SDF-1 expression reached a flat stage, and 24 hour was the bets time to study the dose dependence. ②Both the mRNA and protein expression level analyses demonstrated that the expression of SDF-1 in NRK-52E cells treated with 2 ?g/L TGF-?1 for 24 hours was significantly higher than that in normal control group (P