Effect of rhein on expression of thrombospondin-1and transforming growth factorβ_1 in NRK52E cells stimulated by high glucose

Objective:To investigate the effect of rhein on expression of thrombospondin-1 and transforming growth factor β1 in NRK52E cells stimulated by high glucose..Methods:NRK52E cells were stimulated by different concentrations of rhein(10、20、40ug/ml) in the presence of high glucose(30mmol/L),untreated NRK52E cells were acted as normal control group,NRK52E cells were stimulated by high glucose(30mmol/L) were acted as positive group.Real Time PCR was used to detect the expression of TSP1mRNA and TGF-β1 mRNA.Western blot analysis was used to detect the expression of TSP1 and TGF-β1 protein.ELISA was used to detect the levels of total and active TGF-β1.Results:The expression of TSP1 mRNA(3.34±0.18 vs 1.12±0.13;P20.05)and TGF-β1 mRNA(3.52±0.22 vs 1.24±0.09;P20.05)were up-regulated obviously when stimulated by high glucose(30mmol/L).The protein expression of TSP1(0.5184±0.0736 vs 0.1204±0.0376;P20.01)and TGF-β1(0.5698±0.0324 vs 0.1032±0.0322;P20.01)were also up-regulated simultaneously when stimulated by high glucose(30mmol/L).The expression of TSP1 and TGF-β1 incuced by high glucose(30mmol/L)were down-regulated markedly with 20ug/ml and 40ug/ml of rhein in mRNA and protein levels(P20.05,P20.01).And rhein(20ug/ml and 40ug/ml) could reduce the levels of total and active TGF-β1(P20.05,P20.01).Conclusion: Rhein can inhibit the expression of TSP1 and TGF-β1 in mRNA and protein levels and down-regulate the levels of total and active TGF-β1.