大黄素对TGF-β1诱导的NRK-49F细胞增殖的影响

Effects of Emodin on Proliferation of NRK-49F Cells Induced by TGF-β1

目的:研究大黄素对TGF-β1诱导的肾成纤维细胞株(NRK-49F)增殖以及降低FN(纤连蛋白),Col-I(I型胶原蛋白),Smad2/3蛋白及基因表达的机制。方法:采用CCK8法检测不同浓度大黄素对NRK-49F细胞增殖情况的影响;终浓度为20、40、80μmol/L的大黄素处理NRK-49F细胞30min后,模型组和观察组均以TGF-β1以5ng/mL的终浓度刺激24h,并收集细胞,蛋白质免疫印迹技术(Westernblot)检测FN,Col-I,Smad2/3蛋白表达水平,实时荧光定量PCR(Real-timePCR)检测FN,Col-I,Smad2/3mRNA表达。结果:模型组FN,Col-I,Smad2/3蛋白和基因的表达明显增加,大黄素含药血清组能抑制NRK-49F细胞增殖,且抑制FN,Col-I,Smad2/3蛋白和的表达(P<0.05)。大黄素浓度越高,抑制作用越明显,成一定的量效关系。结论:大黄素干预后,可改善肾纤维化程度,保护NRK-49F细胞模型损伤。

Objective: To investigate the mechanism of emodin on the proliferation of TGF-β1 induced renal fibroblasts(NRK-49F)and the reduction of protein and gene expressions of FN(fibronectin),Col-I(typeⅠcollagen)and Smad2/3. Methods: The effects of different concentrations of emodin on the proliferation of NRK-49F cells were detected by CCK8 assay.NRK-49F cells were treated with emodin at a final concentrations of 20,40,80 μmol/L for 30 min.Both the model group and the treatment group were stimulated with TGF-β1 at a final concentration of 5 ng/mL for 24 h,and the cells were collected.The expression levels of FN,Col-I and Smad2/3 protein were detected by Western blot,and the expressions of FN,Col-I and Smad2/3 mRNA were detected by real-time PCR. Results: The expression of FN,Col-I,Smad2/3 protein and gene in the model group was significantly increased.The emodin-containing serum group inhibited the proliferation of NRK-49F cells and inhibited the expression of FN,Col-I,Smad2/3 protein( P <0.05).The higher the concentration of emodin was,the more obvious the inhibition was.Emodin concentration and inhibition were dose-effect relationships. Conclusion: After emodin intervention,it can improve the degree of renal fibrosis and protect the NRK-49F cell model injury.