姜黄素对TGF-β₁诱导的人肾小管上皮细胞转分化影响的实验研究

【作者】 李彧； 李健； 李亚东； 杨丽霞； 王谦； 牛建昭；

【机构】 北京中医药大学；

【摘要】 目的:通过研究姜黄素（curcumin,Cur）对转化生长因子-β₁（TGF-β₁）诱导人肾小管上皮细胞（HK-2）转分化及分泌细胞外基质（ECM）成分的影响,探讨姜黄素在防治肾小管-间质纤维化方面可能的作用机制。方法:应用不同浓度 Cur 处理经 TGF-β₁诱导活化的 HK-2细胞,通过显微镜观察细胞形态改变,免疫组化技术检测α-平滑肌肌动蛋白（α-SMA）和 E-钙粘蛋白（E-cadherin）的表达:应用酶联免疫吸附法（ELISA）检测细胞培养上清Ⅰ型胶原（Col Ⅰ）、Ⅲ型胶原（Col Ⅲ）和纤连蛋白（FN）的分泌。结果:①正常 HK-2细胞表达 E-cadherin,不表达α-SMA,经 TGF-β₁刺激后细胞表型发生转变, E-cadherin 的表达明显减弱,α-SMA 表达明显增强;②不同浓度 Cur 组与单纯 TGF-β₁刺激组相比α-SMA 的表达有所减弱,而 E-cadherin 的表达增强:③不同浓度 Cur 抑制了 Col Ⅰ、 Col Ⅲ和 FN 的分泌,与单纯 TGF-β₁刺激组相比有显著性差异（p<0.05）。结论:姜黄素能抑制 TGF-β₁诱导人肾小管上皮细胞转分化及细胞外基质成分 Col Ⅰ、Col Ⅲ和 FN 的合成,在一定程度上具有防治肾小管-间质纤维化的作用。更多还原

【Abstract】 Objective:Tubular epithelial-myofibroblast transdifferentiation may be one of the important mechanisms of interstitial fibrosis in progressive renal diseases.The aim of this study is to examine effect of curcumin on transdifferentiation of human tubular epithelial cells （HK-2）and secretion ofextracellular material which were type Ⅰ collagen,type Ⅲ collagen and fibronectin in culture medium supernatant of HK-2 cells induced by transforming growth factor-β₁（TGF-β₁）.And further to explore the probability of curcumin on the prevention and treatment of renal fibrosis.Method:The human tubular epithelial cells（HK-2）were cultured for 24 hours in different conditions:①Free serum as control group;②Treated with TGF-β₁（10 ng/ml）;③Treated with curcumin at different concentration（1、5、10βmoL/L）plus TGF-β₁（10 ng/ml）.HK-2 cells morphology was observed through the microscope,and the expression of α-SMA and E-cadherin of HK-2 were assessed with indirect immunohistochemical staining.The concentration of type Ⅰ collagen,type Ⅲ collagen and FN in culture medium supernatant were detected by ELISA.Result:HK-2 cells morphology can be maintained by curcumin.The expression of α-SMA of HK-2 cultured with TGF-β₁ was much notable than the control,and significantly decreased in HK-2 cultured with TGF-β₁ plus curcumin.The expression of E-cadherin was markedly decreased in HK-2 treated with TGF-β₁,but not changed in HK-2 treated with TGF-β₁ plus curcumin.The secretion of type Ⅰ collagen,type Ⅲ collagen and FN were markedly inhibited in culture medium supernatant of HK-2 cells cultured with TGF-β₁ plus curcumin compared with only TGF-β₁（P<0.05）.Conclution:Curcumin at given concentrations may inhibit tubular epithelial-myofibroblast transdifferentiation,reduce synthesis and secretion of type Ⅰ collagen,type Ⅲ collagen and FN of HK-2 induced by TGF-β₁ in a dose-dependent manner.Our study showed that curcumin could prevent the development of renal fibrosis to a certain extent.更多还原