二甲双胍对TGF-β_(1)诱导的大鼠肺泡上皮Ⅱ型细胞间质转分化的影响  

Effects of metformin on epithelial-mesenchymal transition of rat alveolar epithelial type Ⅱ cells induced by TGF-β_(1)

作　　者：郝伟[1] 左东泽 张俊秀[2] 蒋莉莉[2] 熊莺[2] 杨解人[2] HAO Wei;ZUO Dongze;ZHANG Junxiu;JIANG Lili;XIONG Ying;YANG Jieren(Kinesiology Experimental Training Center,Wannan Medical College,Wuhu 241000,Anhui,China;Department of Pharmacology,Wannan Medical College,Wuhu 241000,Anhui,China)

机构地区：[1]皖南医学院机能学实验实训中心,安徽芜湖241000 [2]皖南医学院药理学教研室,安徽芜湖241000

出　　处：《中国临床药理学与治疗学》2021年第7期744-752,共9页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：安徽省高校优秀青年人才支持计划重点项目资助(gxyqZD2016170);2016年皖南医学院校中青年科研基金资助(WK201602)。

摘　　要：目的:观察二甲双胍(metformin,Met)对大鼠肺泡上皮Ⅱ型细胞(rat alveolar epithelial type Ⅱ cells,RLE-6TN)间质转分化(endothelial-mesenchymal transition,EMT)的影响及其可能的机制。方法:实验分为6组:Control组;转化生长因子-β_(1)(transforming growth factor-β_(1),TGF-β_(1))组:细胞用TGF-β_(1)(5 ng/mL)处理48 h;TGF-β_(1)+Met(1、10、100μmol/L)组:细胞先用Met(1、10、100μmol/L)预处理1 h,再用TGF-β_(1)(5 ng/mL)处理48 h;Met(100μmol/L)组:细胞用Met(100μmol/L)处理48 h。CCK-8法检测细胞增殖。RT-PCR检测细胞α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、波形蛋白(vimentin)、E钙粘附蛋白(E-Cadherin)、紧密连接蛋白-1(zonulaoccludens,ZO-1)、I型胶原(collagen I)、Ⅲ型胶原(collagen Ⅲ)的mRNA表达。Western blotting检测α-SMA、vimentin、E-Cadherin、ZO-1、collagen I、collagen Ⅲ的蛋白表达及Smad2/3和细胞外调节蛋白激酶1/2(extracellular regulated protein kinases 1/2,ERK1/2)的蛋白磷酸化水平。结果:与TGF-β_(1)组比,二甲双胍(10、100μmol/L)可显著抑制TGF-β_(1)诱导的RLE-6TN细胞增殖(P<0.05或P<0.01),降低α-SMA、vimentin、collagen I、collagen Ⅲ的mRNA和蛋白表达及Smad2/3和ERK1/2的蛋白磷酸化水平(P<0.05或P<0.01),增加E-cadherin和ZO-1的mRNA和蛋白表达(P<0.05或P<0.01)。结论:二甲双胍(10、100μmol/L)对TGF-β_(1)诱导的RLE-6TN细胞EMT具有一定的抑制作用,其机制可能与抑制Smad2/3和ERK1/2的磷酸化有关。AIM:To observe the effect of metformin(Met)on the endothelial-mesenchymal transition(EMT)of rat alveolar epithelial type Ⅱ cells and its mechanism.METHODS:The RLE-6TN cells were divided into 6 groups as follows:Control group;transforming growth factor-β_(1)(TGF-β_(1))group in which cells were incubated with TGF-β_(1)(5 ng/mL)for 48 h;TGF-β_(1)+Met(1,10,100μmol/L)group in which cells were pre-treated with Met(1,10,100μmol/L)for 1 h,and then subjected to TGF-β_(1)(5 ng/mL)for 48 h;Met(100μmol/L)group in which cells were incubated with Met(100μmol/L)for 48 h.CCK-8 method was used to detect the cell proliferation.RT-PCR detectedα-smooth muscle actin(α-SMA),vimentin,E-Cadherin,tight junction protein-1(ZO-1),collagen I and collagen Ⅲ mRNA expression.Western blotting was used to detect the protein expression ofα-SMA,vimentin,E-Cadherin,ZO-1,collagen I,collagen Ⅲ and levels of phosphorylated Smad2/3 and extracellular regulated protein kinases(ERK1/2).RESULTS:Metformin significantly inhibited the proliferation of RLE-6TN cells induced by TGF-β_(1)(P<0.05 or P<0.01),reduced the mRNA and protein expression ofα-SMA,vimentin,collagen I,collagen Ⅲ,and decreased the protein phosphorylation of Smad2/3 and ERK1/2 Level(P<0.05 or P<0.01)when compared with the TGF-β_(1) group.Conversely,metformin treatment increased the mRNA and protein expression of E-cadherin and ZO-1(P<0.05 or P<0.01)compared with the TGF-β_(1) group.CONCLUSION:Metformin(10,100μmol/L)exhibits certain inhibitory effect on the EMT of RLE-6TN cells induced by TGF-β_(1),and its beneficial effects may be associated with the inhibition of the phosphorylation of Smad2/3 and ERK1/2.

关 键 词：二甲双胍 肺纤维化 内皮-间质转化 转化生长因子-β_(1) 

分 类 号：R965.2[医药卫生—药理学]