目的 探討貝那普利對(duì)氧化低密度脂蛋白（ox-LDL）誘導(dǎo)的腎小管上皮NRK-52E細(xì)胞間質(zhì)轉(zhuǎn)化（EMT）及轉(zhuǎn)化生長(zhǎng)因子-β（TGF-β）/Smad通路的影響。方法 體外培養(yǎng)NRK-52E細(xì)胞，分為對(duì)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(xué)顯微鏡觀察各組NRK-52E細(xì)胞表型變化；免疫熒光染色法檢測(cè)EMT標(biāo)志性蛋白α-SMA、E-cadherin熒光強(qiáng)度；免疫印跡法檢測(cè)各組NRK-52E細(xì)胞中α-SMA、E-cadherin蛋白及TGF-β/Smad通路蛋白表達(dá)水平。結(jié)果 與對(duì)照組比較，ox-LDL組梭形或不規(guī)則形細(xì)胞增多，α-SMA熒光強(qiáng)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(dá)強(qiáng)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(xì)胞表型改變明顯減輕，α-SMA熒光表達(dá)強(qiáng)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(dá)強(qiáng)度及蛋白量明顯升高（P<0.05）；TGF-β1組細(xì)胞α-SMA熒光表達(dá)強(qiáng)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(dá)強(qiáng)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(xì)胞α-SMA熒光表達(dá)強(qiáng)度明顯增強(qiáng)，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(dá)強(qiáng)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(xì)胞α-SMA熒光表達(dá)強(qiáng)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(dá)強(qiáng)度及蛋白量明顯升高（P<0.05）。結(jié)論 貝那普利可逆轉(zhuǎn)ox-LDL誘導(dǎo)的腎小管上皮NRK-52E細(xì)胞間質(zhì)轉(zhuǎn)化的發(fā)生，機(jī)制可能是通過(guò)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(xué)科技攻關(guān)聯(lián)合共建項(xiàng)目（LHJG20190569）