Effect of transforming growth factor beta-1 on expression of connective tissue growth factor in rat leptomeningeal mesothelial cells

Objective To investigate the role of transforming growth factor beta-1( TGF-β 1) on the expression of connective tissue growth factor( CTGF) in rat leptomeningeal mesothelial cells( RLMCs). Methods RLMCs cultured in vitro were divided into 4 groups:( 1) control group;( 2) 1 ng / ml TGF-β 1 group;( 3) 2 ng / ml TGF-β 1 group;( 4) 4 ng / ml TGF-β 1 group; Cultured RLMCs were incubated in TGF-β 1 at different density for 6 h,12 h and 24 h,the expression of CTGF mRNA was detected by reverse transcription-polymerase chain reaction( RT-PCR),the expression of CTGF protein was examined by Western blot analysis. Results are expressed as mean ± SD. Statistical significance during doses was determined by one factor analysis of variance( one-way ANOVA) and followed by Fishers least significant difference( LSD) test for multiple comparison. P 0. 05 was considered significant. Results RT-PCR: The dose-response of TGF-β 1 elevated the CTGF mRNA: The expression of CTGF mRNA stimulated by incubation of TGF-β 1 1 ng / ml group,2 ng / ml group,4 ng / ml group were significantly higher than that in control group( F 6h = 46. 549,F 12h = 287. 098,F 24h = 109. 202,P 0. 001),and TGF-β 1 performed in a dose-dependent manner; TGF-β 1 mostly increased CTGF mRNA after 12 h treatment. Western-blotting: Compare to control group,CTGF protein was obviously upregulated by TGF-β 1 at all the tree doses and time points( F 6h = 52. 988,F 12h = 95. 331, F 24h = 157. 107,P 0. 001). The regulation showed in a dose-dependent manner and was extremely significant at 6 h. Conclusions TGF-β 1 can activate CTGF pathway in RLMCs. TGF-β 1 can up-regulate CTGF expression in RLMCs and the effect became more and more prominant with increased cell density. CTGF may be further investigated as the target of suppressing meninges fibrosis.