目的 探討貝那普利對氧化低密度脂蛋白（ox-LDL）誘導的腎小管上皮NRK-52E細胞間質(zhì)轉化（EMT）及轉化生長因子-β（TGF-β）/Smad通路的影響。方法 體外培養(yǎng)NRK-52E細胞，分為對照組、ox-LDL組（50μg/mL ox-LDL）、貝那普利組（50 μg/mL ox-LDL＋10 μmol/L貝那普利）、TGF-β1組（50μg/mL ox-LDL＋5 ng/mL TGF-β1）、貝那普利＋TGF-β1組（50 μg/mL ox-LDL＋5 ng/mL TGF-β1＋10 μmol/L貝那普利）。高倍光學顯微鏡觀察各組NRK-52E細胞表型變化；免疫熒光染色法檢測EMT標志性蛋白α-SMA、E-cadherin熒光強度；免疫印跡法檢測各組NRK-52E細胞中α-SMA、E-cadherin蛋白及TGF-β/Smad通路蛋白表達水平。結果 與對照組比較，ox-LDL組梭形或不規(guī)則形細胞增多，α-SMA熒光強度及α-SMA、TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白量顯著升高（P<0.05）；E-cadherin熒光表達強度減弱，蛋白量明顯降低（P<0.05）。與ox-LDL組比較，貝那普利組細胞表型改變明顯減輕，α-SMA熒光表達強度明顯減弱，α-SMA、TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白量顯著降低（P<0.05），E-cadherin熒光表達強度及蛋白量明顯升高（P<0.05）；TGF-β1組細胞α-SMA熒光表達強度及α-SMA、TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白量顯著升高（P<0.05），E-cadherin熒光表達強度及蛋白量明顯降低（P<0.05）。與貝那普利組比較，貝那普利＋TGF-β1組細胞α-SMA熒光表達強度明顯增強，α-SMA、TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白量顯著升高（P<0.05），E-cadherin熒光表達強度及蛋白量明顯降低（P<0.05）。與TGF-β1組比較，貝那普利＋TGF-β1組細胞α-SMA熒光表達強度及α-SMA、TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白量顯著降低（P<0.05），E-cadherin熒光表達強度及蛋白量明顯升高（P<0.05）。結論 貝那普利可逆轉ox-LDL誘導的腎小管上皮NRK-52E細胞間質(zhì)轉化的發(fā)生，機制可能是通過抑制TGF-β/Smad通路活化而發(fā)揮作用。

Objective To investigate the effects of benazepril on ox-LDL-induced epithelial mesenchymal transition (EMT) and TGF-β/Smad pathway in renal tubular epithelial NRK-52E cells. Methods NRK-52E cells were cultured in vitro and divided into control group, ox-LDL group (50 μg/mL ox-LDL), benazepril group (50 μg/mL ox-LDL + 10 μmol/L benazepril), TGF-β1 group (50 μg/mL ox-LDL + 5 ng/mL TGF-β1), and benazepril + TGF-β1 group (50 μg/mL ox-LDL + 5 ng/mL TGF-β1 + 10 μmol/L benazepril). The phenotypic changes of NRK-52E cells were observed by high power optical microscope, the fluorescence intensity of α-SMA and E-cadherin was detected by immunofluorescence staining, Western blot was used to detect the expression of α-SMA, E-cadherin and TGF-β/Smad pathway protein in NRK-52E cells. Results Compared with the control group, the number of spindle or irregular cells in ox-LDL group was more, the fluorescence intensity of α-SMA, α-SMA, TGF-β1, p-Smad2/Smad2, p-Smad3/Smad3 protein were significantly higher (P < 0.05), the fluorescence intensity and protein quantity of E-cadherin were significantly weaker and lower respectively (P < 0.05). Compared with those in ox-LDL group, the phenotypic changes of benazepril group were significantly reduced, the fluorescence intensity of α-SMA was significantly weaker, the protein levels of α-SMA, TGF-β1, p-Smad2/Smad2, and p-Smad3/Smad3 were significantly lower (P < 0.05), the fluorescence intensity and protein quantity of E-cadherin were significantly stronger and higher respectively (P < 0.05). In the TGF-β1 group, the fluorescence intensity of α-SMA, α-SMA, TGF-β1, p-Smad2/Smad2, p-Smad3/Smad3 protein were significantly higher (P < 0.05), the fluorescence intensity and protein quantity of E-cadherin were significantly weaker and lower respectively (P < 0.05). Compared with benazepril group, the fluorescence intensity of α-SMA in benazepril + TGF-β group was significantly stronger, the protein levels of α-SMA, TGF-β1, p-Smad2/Smad2, and p-Smad3/Smad3 were significantly higher (P < 0.05), the fluorescence intensity and protein quantity of E-cadherin were significantly weaker and lower respectively (P < 0.05). Compared with those in TGF-β1 group, the fluorescence intensity of α-SMA, α-SMA, TGF-β1, p-Smad2/Smad2, p-Smad3/Smad3 protein in the benazepril + TGF-β1 group were significantly lower (P < 0.05), the fluorescence intensity and protein quantity of E-cadherin were significantly stronger and higher respectively (P < 0.05). Conclusion Benazepril can reverse ox-LDL-induced EMT in NRK-52E cells, it may play a role by inhibiting the activation of TGF-β/Smad pathway.

R965

河南省醫(yī)學科技攻關聯(lián)合共建項目（LHJG20190569）