Transforming growth factor beta 1 induces the transdifferentiation of human renal tubular epithelial cells: the influence of Notch1 receptor specific inhibitor

doi:10.3969/j.issn.2095-4344.2014.51.012

Abstract

Abstract:

BACKGROUND: In the pathological state, a variety of cells 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.

OBJECTIVE: To explore whether the γ-secretase inhibitor DAPT, specific inhibitor of Notch1 receptor can effectively block, completely or partially reverse the tubular epithelial-mesenchymal transition induced by TGF-β1.

METHODS: Normal human kidney epithelial cell lines (HK-2) cultured in vitro were used to establish in vitro model of tubular epithelial-mesenchymal transition, and then divided into blank control group, 10 μg/L TGF-β1 group, 10 μg/L TGF-β1+5 μmol/L DAPT inhibited group, 10 μg/L TGF-β1+5 μmol/L DAPT partially 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.

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 partially delayed group, the mRNA and protein expressions of a-smooth muscle actin were increased at 12 hours (P < 0.05), and then gradually decreased (P < 0.05); the expression of E-cadherin protein began to decrease at 24 hours (P < 0.05), and then increased gradually; the mRNA expression of E-cadherin was similar in the TGF-β1+DAPT partially 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 partially but not completely block and reverse the tubular epithelial-mesenchymal transition by TGF-β1.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: transforming growth factor beta1, actins, epithelial-mesenchymal transition

CLC Number:

R318

Zeng Hong, Zhou Yi, Yao Guo-yuan. Transforming growth factor beta 1 induces the transdifferentiation of human renal tubular epithelial cells: the influence of Notch1 receptor specific inhibitor[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(51): 8261-8268.

Figures/Tables 7

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