雷公藤内酯醇干预高糖诱导足细胞转分化作用机制的研究  

Effect of triptolide on epithelial to mesenchymal transition(EMT) of mouse podocytes induced by high glucose in vitro

作　　者：叶迅[1] 侯鹏超[1] 朱伶俐[1] 洪郁芝[1] 

机构地区：[1]杭州市中医院内分泌代谢科,310007

出　　处：《浙江医学》2015年第17期1428-1431,1459,I0003,共6页Zhejiang Medical Journal

基　　金：浙江省中医药科学研究基金计划(A类)(2012ZA097);杭州市科委医疗卫生及重点专科专病科研攻关专项(20130733Q16)

摘　　要：目的观察不同浓度雷公藤内酯醇(TP)体外干预高糖环境下小鼠足细胞上皮细胞向间充质细胞转分化(EMT)及TGF-β/Smad信号通路的调节作用,探讨其保护足细胞损伤的可能机制。方法将培养成熟的小鼠足细胞随机分为25mmol/L葡萄糖培养液干预组(高糖组),5mmol/L葡萄糖培养液干预组(对照组)和在25mmol/L葡萄糖培养液中分别加入浓度为8、16、32ng/ml的TP干预组(低、中、高TP组)。体外培养48h后,倒置显微镜下观察足细胞形态变化,采用RT-PCR和Western-blot技术分别检测各组足细胞NEPH1、desmin、TGF-β_1和Smad7的mRNA和蛋白表达变化。结果高糖组和高、中、低TP组的小鼠足细胞NEPH1和Smad7 mRNA和蛋白的表达水平均较对照组低,desmin和TGF-β_1的mRNA和蛋白的表达水平均较对照组高,差异有统计学意义(P<0.05或0.01)。高、中、低TP组足细胞NEPH_1和Smad7的mRNA和蛋白的表达水平均高于高糖组,desmin和TGF-β_1的mRNA和蛋白的表达水平均低于高糖组,差异均有统计学意义(P<0.05或0.01);中TP组NEPH1和Smad7的mRNA和蛋白的表达水平均高于低TP组和高TP组,desmin和TGF-β_1的mRNA和蛋白的表达水平均低于低TP组和高TP组(均P<0.01)。结论TP可能通过抑制TGF-β/Smad信号通路缓解高糖诱导的足细胞转分化。Objective To investigate the effect of triptolide（TP） on epithelial to mesenchymal transition（EMT） of mouse podocytes induced by high glucose in vitro. Methods Matured mouse podocytes were cultured and randomly divided into control group（5mmol/L D-glucose）, high glucose group（25mmol/L D-glucose） and TP-treated groups（25mmol/L D-glucose with 8ng/ml, 16ng/ml or 32ng/ml triptolide）. Morphological changes of podocytes were observed with inverted microscope, the ex- pressions of NEPH1, desmin, TGF- β, Smad7 mRNA and proteins were detected by RT-PCR and Western blot, respectively. Results Matured podocytes incubated with high glucose for 24h were converted from arborized into cobblestone-like cells, while the cobblestone-like cells converted back to arborized cells after cultured with different doses of triptolide in varying degrees. Compared with the control group, the expression of NEPH1 and Smad7 cultured in high glucose was significantly decreased（P〈 0.01）; meanwhile, the expression of desmin and TGF-β1 was significantly increased in comparison with those of control group （P〈0.01）. The inhibited expressions of NEPH1 and Smad7 were significantly up-regulated in all TP groups in varying degrees （P〈0.05 or P〈0.01）, while those of desmin and TGF-β1 were significantly down-regulated compared with high glucose group （P〈0.05 or P〈0.01）; the changes of expression were more markedly in 16 ng/ml triptolide group （P〈0.01）. Conclusion The transdifferentiation of mouse podocytes cultured in high glucose can be attenuated by triptolide via inhibiting the TGF- β/Smad signal pathway.

关 键 词：雷公藤内酯醇 糖尿病肾病 足细胞 转分化 

分 类 号：R285.5[医药卫生—中药学]