外源性结缔组织生长因子对肾小管上皮细胞的转分化和胶原合成效应

Transdifferentiation and collagen-synthesis effects of exogenous connective tissue growth factor on renal tubular epithelial cells in vitro

摘要:

目的：大量研究表明内源性结缔组织生长因子(CTGF)在肾小管间质纤维化(TIF)进程中发挥了重要作用,为进一步阐明外源性CTGF的作用机制,该实验探讨了重组人结缔组织生长因子(rhCTGF)刺激对人肾小管上皮细胞(HK2)转分化和胶原合成的作用。方法：rhCTGF 5 ng/mL刺激HK2细胞,用倒置显微镜观察细胞形态学变化,同时在刺激后0,3,6,12,24,48 h各点收集细胞,RT-PCR检测上皮细胞钙粘蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、胶原Iα1(Col Iα1)和胶原IVα1(Col IVα1)mRNA表达变化。结果：rhCTGF 5 ng/mL刺激使HK2细胞形态由椭圆形转为梭形,下调E-cadherin mRNA表达,上调-αSMA mRNA表达,但对Col Iα1和Col IVα1mRNA表达没有影响。结论：外源性CTGF能导致HK2细胞转分化,但不能促进其胶原合成。

Abstract:

OBJECTIVE: To investigate the effects of recombinal human connective tissue growth factor(rhCTGF) stimulation on epithelial-myofibroblast transdifferentiation (EMT) and collagen-synthesis in human renal tubular epithelial cell line(HK2) in vitro. METHODS: The cultured HK2 cells were stimulated with rhCTGF of 5 ng/mL.The morphological changes were observed under an inverted microscope. The cells were collected at 0, 3, 6, 12, 24 and 48 hrs after rhCTGF stimulation. The expression of E-cadherin, α-smooth muscle actin (α-SMA), collagen Iα_1 (Col Iα_1) and collagen IVα_1(Col IVα_1) mRNAs were detected by RT-PCR. RESULTS: rhCTGF stimulation changed the HK2 cell appearance from oval to fusiform, down-regulated the E-cadherin mRNA expression and up-regulated α-SMA mRNA expression, but had no effects the Col Iα_1 and Col IVα_1 mRNA expression. CONCLUSIONS: Exogenous CTGF can mediate the EMT but has no collagen-synthesis effects on HK2 cells.