摘要
Abstract
BACKGROUND:In the pathological state, a variety of cels can be involved in the tubular epithelial-mesenchymal transition into myofibroblasts mediated by transforming growth factor-β1 (TGF-β1), thereby accelerating the progress of renal tubular interstitial fibrosis. <br> OBJECTIVE:To explore whether the γ-secretase inhibitor DAPT, specific inhibitor of Notch1 receptor can effectively block, completely or partialy reverse the tubular epithelial-mesenchymal transition induced by TGF-β1. <br> METHODS: Normal human kidney epithelial cel lines (HK-2) culturedin vitro were used to establishin vitro model of tubular epithelial-mesenchymal transition, and then divided into blank control group, 10μg/L TGF-β1 group, 10 μg/L TGF-β1+5 μmol/L DAPT inhibited group, 10 μg/L TGF-β1+5 μmol/L DAPT partialy delayed group, 10 μg/L TGF-β1+5 μmol/L DAPT delayed group. After 12, 24, 48 and 72 hours, the HK-2 morphologic changes were observed by an inverted phase contrast microscope; the expressions of a-smooth muscle actin and E-cadherin at mRNA and protein levels were examined respectively by immunohistochemistry and RT-PCR. <br> RESULTS AND CONCLUSION: (1) Compared with the blank control group, the mRNA and protein expressions of a-smooth muscle actin and E-cadherin were respectively increased (P < 0.05) and reduced (P < 0.05) significantly at 12, 24, 48 and 72 hours after intervention. (2) There was no difference in the mRNA and protein expression of a-smooth muscle actin and E-cadherin between the blank control group and TGF-β1+DAPT inhibited group (P > 0.05). (3) In the TGF-β1+DAPT partialy delayed group, the mRNA and protein expressions of a-smooth muscle actin were increased at 12 hours (P < 0.05), and then gradualy decreased (P < 0.05); the expression of E-cadherin protein began to decrease at 24 hours (P < 0.05), and then increased gradualy; the mRNA expression of E-cadherin was similar in the TGF-β1+DAPT partialy delayed group and blank control group at different time points after intervention; the mRNA and protein expressions of a-smooth muscle actin and E-cadherin showed no difference from the blank control group at 72 hours after intervention (P > 0.05). (4) Compared with the blank control group, the expressions of a-smooth muscle actin and E-cadherin were respectively increased (P < 0.05) and reduced (P < 0.05) significantly after intervention in the TGF-β1+DAPT delayed group, but there was no difference in the expression of E-cadherin at 72 hours after intervention between the two groups. These findings indicate that DAPT can partialy but not completely block and reverse the tubular epithelial-mesenchymal transition by TGF-β1.关键词
组织构建/组织工程/细胞转分化/转化生长因子β1/Notch1受体/γ-分泌酶抑制剂DAPT/E-钙粘连素/α-平滑肌肌动蛋白Key words
Subject headings:transforming growth factor beta1/actins/epithelial-mesenchymal transition分类
医药卫生
