【摘要】 目的：探讨发酵红参总皂苷（FRGTS）对高糖诱导下肾小管上皮细胞发生上皮细胞-间充质转化（EMT）的抑制作用，并阐明其作用机制。方法：将大鼠肾小管上皮NRK-52E细胞分为正常对照组(5.5 mmol·L^-1D-葡萄糖)、高糖组(30.0 mmol·L^-1D-葡萄糖)、沉默信息调节因子1（SIRT1）抑制剂EX527组(30.0 mmol·L^-1D-葡萄糖+10.0μmol·L^-1.EX527)、 FRGTS组(30.0 mmol·L^-1D-葡萄糖+25 mg·L^-1FRGTS)和EX527+FRGTS组(30.0 mmol·L^-1D-葡萄糖+10μmol·L^-1EX527+25 mg·L^-1FRGTS)。采用免疫荧光法检测各组细胞中E-钙黏蛋白（E-cadherin）和α-平滑肌肌动蛋白（α-SMA）蛋白表达水平，实时荧光定量PCR （RT-qPCR）法检测各组细胞中E-cadherin、α-SMA和SIRT1 mRNA表达水平，ELISA法检测培养上清液中Ⅰ型胶原（ColⅠ）水平，Western blotting法检测各组细胞中SIRT1、转化生长因子β1 （TGF-β1）和Smad3蛋白表达水平。结结果果：高糖培养48 h后，与正常对照组比较，高糖组NRK-52E细胞中E-cadherin蛋白表达水平和mRNA表达水平降低（P<0.01）,α-SMA蛋白表达水平和mRNA表达水平升高（P<0.01）,NRK-52E细胞上清液中ColⅠ水平升高（P<0.01）,SIRT1 mRNA和蛋白表达水平均明显降低（P<0.01）,TGF-β1和Smad3蛋白表达水平升高（P<0.01）；与高糖组比较，FRGTS组NRK-52E细胞中E-cadherin mRNA表达水平升高（P<0.01）,α-SMA mRNA表达水平降低（P<0.05）,NRK-52E细胞上清液中ColⅠ水平降低（P<0.05）, SIRT1 mRNA和蛋白表达水平均升高（P<0.05或P<0.01）,TGF-β1和Smad3蛋白表达水平降低（P<0.05或P<0.01）；与FRGTS组比较，EX527+FRGTS组NRK-52E细胞中E-cadherin mRNA表达水平明显降低（P<0.01）,α-SMA mRNA表达水平明显升高（P<0.05）, NRK-52E细胞上清液中ColⅠ水平升高（P<0.05）, SIRT1 mRNA和蛋白表达水平明显降低（P<0.01）, TGF-β 1和Smad3蛋白表达水平明显升高（P<0.01）。结结论论：FRGTS可通过上调SIRT1表达，进而抑制TGF-β1/Smad信号通路，有效抑制肾小管上皮细胞发生EMT。更多还原

【Abstract】 Objective:To investigate the inhibitory effect of fermented red ginseng total saponins（FRGTS）on the epithelial-mesenchymal transition（EMT）of renal tubular epithelial cells induced by high glucose,and to clarify its mechanisms. Methods:The rat renal tubular epithelial cells NRK-52E were divided into normal control group(5. 5 mmol·L^-1D-glucose),high glucose group(30. 0 mmol·L^-1D-glucose),silent information regulator 1（SIRT1）inhibitor EX527 group(30. 0 mmol·L^-1D-glucose+10 μmol·L^-1EX527),FRGTS group(30. 0 mmol·L^-1D-glucose+25 mg·L^-1FRGTS)and EX527+FRGTS group(30. 0 mmol·L^-1Immunofluorescence method was used to detect the expression levels of E-cadherin and α-smooth muscle actin（α-SMA） protein in the cells in various groups. Real-time fluorescence quantitative PCR（RTqPCR） method was used to detect the expression levels of E-cadherin,α-SMA and SIRT1 mRNA,ELISA method was used to detect the levels of collagen type Ⅰ （Col Ⅰ） in culture supernatant,and Western blotting method was used to detect the expression levels of SIRT1,transforming growth factor-β1（TGF-β1）and Smad3 proteins in the cells in various groups. Results:After 48 h of high glucose culture,compared with normal control group,the expression levels of E-cadherin mRNA and protein in the NRK-52E cells in high glucose group were decreased（P<0. 01）,the expression levels of α-SMA mRNA and protein were increased（P<0. 01）,the level of Col Ⅰ in culture supernatant was increased（P<0. 01）,the expression levels of SIRT1 mRNA and protein were significantly decreased（P<0. 01）,and the expression levels of TGF-β1 and Smad3 proteins were increased（P<0. 01）. Compared with high glucose group,the expression level of E-cadherin mRNA in the NRK-52E cells in FRGTS group was increased（P<0. 01）,the expression level of α-SMA mRNA was decreased（P<0. 05）,the level of Col Ⅰ in culture supernatant was decreased（P<0. 05）,the expression levels of SIRT1 mRNA and protein were increased（P<0. 05 or P<0. 01）,and the expression levels of TGF-β1 and Smad3 proteins were decreased（P<0. 05 or P<0. 01）. Compared with FRGTS group,the expression level of E-cadherin mRNA in the cells in EX527+FRGTS group was significantly decreased（P<0. 01）,the expression level of α-SMA mRNA was increased（P<0. 05）,the level of Col Ⅰ in culture supernatant was increased（P<0. 05）,the expression levels of SIRT1 mRNA and protein were significantly deceased（P<0. 01）,and the expression levels of TGF-β1 and Smad3 were significantly increased（P<0. 01）. Conclusion:FRGTS can upregulate the expression of SIRT1 and then inhibits TGF-β1/Smad signaling pathway,which effectively inhibits the EMT of renal tubular epithelial cells.更多还原