肝细胞生长因子对转化生长因子诱导肾小管上皮细胞转分化的影响  

Effects of Hepatocyte Growth Factor on TGF-β1 Triggered Tubular Epithelial-Myofibroblast Transdifferentiation by CTGF in vitro

作　　者：钟慧[1] 樊均明[1] 李孜[1] 刘芳[1] 杨立川[1] 冀玲[1] 沙朝晖[1] 马行一[1] 

机构地区：[1]四川大学华西医院肾内科,成都610041

出　　处：《四川大学学报（医学版）》2005年第5期653-656,691,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30170437);国家教委留学回国人员启动基金(C299005)资助

摘　　要：目的应用转化生长因子β1(TGF-β1)诱导肾小管上皮细胞肌纤维母细胞转分化,研究肝细胞生长因子(HGF)对肾小管上皮细胞的表型重塑的作用和机制。方法在正常大鼠肾小管上皮细胞(NRK52E细胞株)培养液中分别加入TGF-β1及HGF,应用倒置相差显微镜、扫描电镜观察细胞形态变化;免疫组织化学方法检测肌纤维母细胞特异性标志物α-平滑肌肌动蛋白(α-SMA)的表达;ELISA法分析测定培养细胞上清液中纤维连结蛋白(FN)的含量;RT-PCR检测结缔组织生长因子(CTGF)mRNA表达。结果1TGF-β1诱导肾小管上皮细胞从原有典型的上皮细胞形态转变为长梭形肌成纤维细胞样形态;α-SMA的表达、上清液FN含量、细胞CTGFmRNA表达明显增加(P<0.05)。2单纯HGF对细胞形态学、α-SMA的表达及FN含量与空白对照组无明显影响;不同浓度HGF组的CTGFmRNA表达较空白对照组均增加(P<0.05)。3HGF能抑制TGF-β1诱导的NRK52E细胞形态学改变;α-SMA的表达、FN水平均比诱导组TGF-β1明显下降(P<0.05);HGF抑制了TGF-β1诱导的CT-GFmRNA表达(P<0.05)。结论HGF能够负性调控TGF-β1诱导的肾小管上皮细胞肌纤维母细胞转分化作用,减少FN分泌;HGF对TEMT的负性调节作用可能是通过减少CTGF表达来实现的。Objective To observe the effects of hepatocyte growth factor （HGF） on TGF-β1 triggered tubular epithelial-myofibroblast transdifferentiation （TEMT） and on the expression of connective tissue growth factor （CTGF）. Methods The morphology of transdifferentiate tubular cells was observed using phase-contrast microscopy and scanning electron microscopy, α-SMA was assessed by immunohistochemistry and semiquantified by mean intergrated opitical density （IOD）. The level of fibronectin （FN） in the culture supernatant was measured by ELISA. CTGF mRNA expression was examined by RT-PCR. Results The TGF-β1-induced TEMT characterized by expression of α-SMA was shown by immunohistochemistry. TGF-β1 was also shown to stimulate the secretion of FN in cultured supernatant and the CTGF mRNA expression of NRK52E cells. There was no statistically significant difference between HGF-treated groups and control group in the result of α-SMA immunostaining and the level of FN, except that CTGF mRNA expression was slightly increased in the HGF-treated groups. The addition of HGF inhibited the TGF-β1-induced TEMT, the secretion of FN, and the CTGF expression of NRK52E cells, there was a significant correlation between the expression of CTGF and the expression of α-SMA. Conclusion HGF could block TEMT and FN secretion triggered by TGF-β1, which implies that HGF could participate in renal interstitial fibrosis as a negative regulator. The negative regulation of transdifferentiation of HGF may be partially achieved by attenuation of CTGF expression.

关 键 词：肝细胞生长因子结缔组织生长因子 转分化 肾间质纤维化 转化生长因子Β1 

分 类 号：R692.6[医药卫生—泌尿科学]