姜黄素对大鼠肾间质成纤维细胞中小分子RNA-21及纤维化相关因子表达的影响

Effects of curcumin on the expression of microRNA-21 and fibrosis-related factors in rat renal interstitial fibroblast cells

目的探讨姜黄素对转化生长因子β1(TGF-β1)诱导的大鼠肾间质成纤维细胞(NRK49F)中小分子RNA-21(miR-21)及纤维化相关因子表达的影响。方法将NRK49F细胞分为正常组、模型组、姜黄素组。模型组以TGF-β1(10μg/L)诱导NRK49F细胞活化,姜黄素组以高、中、低浓度(20、10、5μmol/L)干预24 h和72 h。免疫荧光方法及高内涵成像分析设备检测并分析NRK49F细胞中α-平滑肌肌动蛋白(α-SMA)表达的变化;RT-PCR方法检测NRK49F细胞中miR-21及纤维化相关因子COL1A1、COL3A1、α-SMA、Smad3 mRNA表达的变化。结果 TGF-β1作用于NRK49F细胞72 h后,能明显上调细胞中α-SMA蛋白的表达(P<0.01);与模型组相比,姜黄素各组的α-SMA蛋白表达明显下调(P<0.01、P<0.01、P<0.05)。TGF-β1作用于NRK49F细胞24 h后,能明显上调细胞中miR-21、COL1A1、COL3A1、α-SMA、Smad3 mRNA的表达(P<0.01);与模型组相比,姜黄素高、中浓度组可明显抑制miR-21、COL1A1、COL3A1、α-SMA、Smad3 mRNA的表达(P<0.01或P<0.05);姜黄素低浓度组可明显抑制COL1A1、COL3A1、α-SMA mRNA的表达(P<0.01),对miR-21、smad3 mRNA的表达有下调的趋势。结论姜黄素能够有效抑制TGF-β1诱导的NRK49F细胞中纤维化相关因子的基因及蛋白表达,其作用与下调miR-21的表达相关。

Objective To investigate the effects of curcumin on expression of microRNA-21 and fibrosisrelated factors induced by transforming growth factor-beta 1（ TGF-β1） in the renal interstitial fibroblast cells（ NRK49 F） of rats. Methods NRK49 F cells were divided into normal group,model group,and curcumin groups. NRK49 F cells in the model group were activated by TGF-β1（ 10μg/L）. Cells in the curcumin groups were treated with curcumin at high,medium,and low concentration（ 20,10,5μmol/L respectively） for 24 and 72 hours. The protein expression of α-SMA were measured by using immunofluorescence cell staining and cell analyzer. The expression of micro RNA-21 and mRNA of fibrosis-related factors（ COL1 A1,COL1 A2,COL3 A1,ACTA2,Smad3） were measured with RT-PCR. Results After treated with TGF-β1 for 72 hours,the protein expression level of α-SMA increased significantly（ P〈0. 01）; Compared with TGF-β1-treated group, the protein expression level of α-SMA decreased significantly in the curcumin groups（ 20,10,5 μmol/L）（ P〈0. 01,P〈0. 01,P〈0. 05）. After treated with TGF-β1 for 24 hours,the expression level of miR-21,COL1 A1,COL3 A1,α-SMA,and Smad3 mRNA increased significantly（ P〈0. 01）. Compared with TGF-β1-treated group,the expression level of miR-21,COL1 a1,COL3 a1,α-SMA,and Smad3 mRNA decreased significantly in the curcumin groups（ 20,10 μmol/L）（ P〈0. 01,P〈0. 05）; Compared with TGF-β1-treated group,the expression level of COL1 A1,COL3 A1,and α-SMA mRNA decreased significantly（ P〈0. 01） and the expression level of miR-21 and smad3 mRNA showed a downward trend in the 5 μmol/L curcumin group. Conclusion Curcumin down-regulated fibrosis-related gene and protein expression induced by TGF-β1 in rat renal interstitial fibroblasts,which may be related to its down-regulating the expression of miR-21 genes.