黄芪通过c-met调控TGF-β_1诱导的肾小管上皮细胞转分化  

Astragalus Mongholicus May Regulate Rat Renal Tubular Cell Transdifferrentiation Induced by TGF-β_1 through C-met Dependent Pathway

作　　者：肖敏[1] 李会娟[1] 谢席胜[1] 樊均明[1] 

机构地区：[1]四川大学华西医院肾脏内科,成都610041

出　　处：《中国中西医结合肾病杂志》2008年第9期764-768,I0001,共6页Chinese Journal of Integrated Traditional and Western Nephrology

摘　　要：目的:探讨黄芪对TGF-β1诱导的肾小管上皮细胞转分化及细胞外基质分泌的作用及机制。方法:体外培养正常大鼠肾小管上皮细胞(NRK52E),应用倒置相差显微镜观察NRK52E细胞形态学变化;免疫组织化学染色法及实时荧光定量PCR法检测α-平滑肌肌动蛋白(α-SMA),肝细胞生长因子HGF受体(c-met)的表达;ELISA法定量检测细胞上清液中胶原Ⅰ(Col-Ⅰ),胶原Ⅲ(Col-Ⅲ)和纤维黏连蛋白(FN)的水平。结果:TGF-β1可诱导肾小管上皮细胞肌成纤维细胞转分化(TEMT),TGF-β1诱导组细胞肥大、拉长,呈长梭形,α-SMA表达明显增强,Col-Ⅰ、Col-Ⅲ和FN分泌增加(P<0.05)。加入不同浓度黄芪后,细胞形态接近正常肾小管上皮细胞形态,α-SMA表达、Col-Ⅰ、Col-Ⅲ和FN分泌均较TGF-β1诱导组明显抑制(P<0.05),c-met表达较TGF-β1诱导组增加(P<0.05)且呈剂量依赖性。结论:TGF-β1可以诱导肾小管上皮细胞肌成纤维细胞转分化,增加细胞外基质成分Col-Ⅰ、Col-Ⅲ和FN的分泌;黄芪能够抑制TGF-β1诱导的NRK52E细胞转分化以及细胞外基质的分泌;黄芪抑制细胞转分化的机制可能与其增强c-met的表达有关。Objective：To observe the effect and mechanism of Astragalus mongholicus on tubular epithelial - myofibmblast transdifferentiation and the secretion of extra cellular matrix （ECM） induced by TGF β1- Methods： Normal rats＇ tubular epithelial cells NRK52E were cultured in vitro, their morphological changes were observed by inverted phase contrast microscope, a - smooth muscle actin （a-SMA） and the receptor of hepatocyte growth factor（c- met）proteins and genes expression were assessed by irnmunohistochemistry and realtime quantitative polymerase chain reaction. The collagen I , collagen Ⅲ and fibronectin was measured by ELISA. Results：TGF- β1 could induce tubular epithelial - myofibroblast transdifferentiation, showing hypertrophy of cells elongated and fusiforrn - shaped, with significantly enhanced expression of a - SMA and increased secretion of Col - I , Col -Ⅲ and FN（ P 〈 0.05）. After adding of Astragalus mongholieus at different concentrations, the morphology of cells restored close to normal phenotype, with the expression of a- SMA and the secretion of Col - i , Col-Ⅲ and FN significantly inhibited （P 〈 0.05） and the expression of c - met significantly increased （ P 〈 0.05 ） in dose - dependent manner. Conclusion： TGF - β1 can induce tubular ep- ithelial - myofibroblast transdifferentiation and increase the level of collagen I , collagen Ⅲ and fibronectin. Astragalus mongholicus could inhibit transdifferentiation of NRK52E induced by TGF - β1 and decrease the secretion of collagen I , collagen Ⅲ and fibronectin. The effect of Astragalus mongholicus on transdifferentiation may be associated with the increased expression of c met.

关 键 词：黄芪 转化生长因子-β1 肾小管上皮细胞转分化 Α-平滑肌肌动蛋白 肝细胞生长因子受体 胶原Ⅰ胶原Ⅲ 纤维黏连蛋白 

分 类 号：R285.5[医药卫生—中药学]