摘要
目的通过观察高糖环境下给予硫辛酰胺(dl alpha lipoamide,ALM)干预后对细胞氧化应激水平、转化生长因子β激活激酶1(TAK1)及核转录共抑制因子(SnoN)蛋白表达的影响,探讨ALM减轻高糖诱导诱导的肾小管纤维化病变的可能机制。方法将体外培养的肾小管上皮细胞(NRK 52E)分为3组:正常糖培养组(NG)、高糖组培养组(HG)、高糖培养条件下加入ALM刺激组(ALM);采用细胞免疫荧光及Western Blot技术,进行细胞鉴定;氧化应激试剂盒检测体外培养的各组肾小管上皮细胞(NRK 52E cells)氧化应激水平;Western Blot技术检测高糖环境下NRK 52E细胞中,硫辛酰胺对TAK1、SnoN蛋白表达的影响;流式细胞技术和Western Blot技术检测ALM对大鼠肾小管上皮细胞纤维化过程的抑制作用。结果所培养细胞为肾小管上皮细胞并对高糖刺激敏感;高糖条件下给予ALM干预后,T SOD活性增强,MDA水平减少,且TAK1、p TAK1(Thr184/187)及CollagenⅢ蛋白表达水平减少,SnoN及钙黏蛋白(E cadherin)蛋白水平的表达上调。结论高糖环境下给予ALM干预后,肾小管上皮细胞的氧化应激水平降低,伴随TAK1的表达减少和SnoN蛋白表达增加,从而抑制肾小管上皮细胞向间质细胞转分化的发生及细胞外基质沉积,最终延缓肾小管上皮细胞纤维化的发生发展。
Objective To investigatee the effect of dl alpha lipoamide(ALM)on the expression of trans forming growth factor beta activated kinase 1(TAK1)and Ski related novel protein N(SnoN)in renal tubular epi thelial cells under high glucose(HG)treatment,and to explore the role and potential mechanism of ALM in allevi ating the oxidative stress injury.Methods Cell immunofluorescence and Western Blot assay were used to identify renal tubular epithelial cells.Oxidative stress kit was used to detect the level of oxidative stress and the ability of li poamide against oxidative stress in renal tubular epithelial cells(NRK 52E cells).The effects of lipoamide on the stability of TAK1 and SnoN proteins in NRK 52E cells under high glucose were detected.Meanwhile,flowcytome try and Western Blot assay were used to study the inhibitory effect of ALM on rat renal tubular epithelial cell fibro sis.Results The cultured renal tubular epithelial cells were sensitive to hyperglycemic stimulation.The oxidative stress levels were reduced after ALM intervention under high glucose treatment,protein levels of TAK1,p TAK1(Thr184/187)and CollagenIII were decreased.The expression of SnoN and E cadherin protein was up regulated.Conclusion After intervention with ALM and high glucose,the oxidative stress level in renal tubular epithelial cells decreased,accompanied with the decrease of TAK1 expression and the increase of SnoN protein expression,contributing to inhibiting the occurrence of tubular epithelial cells to mesenchymal cells transdifferentiation and the deposition of extracellular matrix,as well as delaying the occurrence and development of renal tubular epithelial fibrosis.
作者
毛彦稳
张小欢
刘慧铭
王圆圆
汤磊
郭兵
MAO Yanwen;ZHANG Xiaohuan;LIU Huiming;WANG Yuanyuan;TANG Lei;GUO Bing(State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Provinical Key Labo ratory of Pharmaceutics,Guizhou Medical University,Guiyang 550025,China)
出处
《实用医学杂志》
CAS
北大核心
2019年第19期2993-2997,3003,共6页
The Journal of Practical Medicine
基金
国家自然科学基金资助项目（编号：81760131）
贵州省科技支撑计划项目（编号：黔科合支撑[2017]2837）
贵阳市科技局基金项目（编号：筑科合同[2017]30-26号）
作者简介
通信作者:郭兵E mail:guobingbs@126.com。
