中国中药杂志

2019, v.44(24) 5457-5464

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

雷公藤甲素抑制NLRP3炎症小体活化改善高糖诱导的足细胞上皮-间充质转分化
Triptolide inhibits NLRP3 inflammasome activation and ameliorates podocyte epithelial-mesenchymal transition induced by high glucose

吴薇;刘不悔;万毅刚;孙伟;刘莹露;王文文;房其军;涂玥;YEE Hong-yun;袁灿灿;万子玥;
WU Wei;LIU Bu-hui;WAN Yi-gang;SUN Wei;LIU Ying-lu;WANG Wen-wen;FANG Qi-jun;TU Yue;YEE Hong-yun;YUAN Can-can;WAN Zi-yue;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital Clinical College of Chinese Medicine and Western Medicine,Nanjing University of Chinese Medicine;Department of Nephrology,the Affiliated Hospital of Nanjing University of Chinese

摘要(Abstract):

基于糖尿病肾病(diabetic kidney disease,DKD)肾组织Nod样受体蛋白3(Nod-like receptor protein 3 inflammasome,NLRP3)炎症小体活化的调控机制而探讨雷公藤有效成分——雷公藤甲素(triptolide,TP)在体外改善高糖(high glucose,HG)诱导足细胞上皮-间充质转分化(epithelial-mesenchymal transition,EMT)的作用。将体外培养的小鼠永生化足细胞分为正常(the normal,N)组、HG组、低剂量TP(the low dose of TP,L-TP)组、高剂量TP(the high dose of TP,H-TP)组以及甘露醇(the mannitol,MNT)组。分别进行不同的干预,即N组加入5 mmol·L~(-1)D-葡萄糖(D-glucose,DG),HG组加入30 mmol·L~(-1)HG,L-TP组加入30 mmol·L~(-1)HG+5μg·L~(-1)TP,H-TP组加入30 mmol·L~(-1)HG+10μg·L~(-1)TP,MNT组加入5 mmol·L~(-1)DG+24. 5mmol·L~(-1)MNT。在干预后的不同时间点(24,48,72 h),首先,观察HG或TP对足细胞增殖活性的影响;其次,检测足细胞上皮细胞标志性分子nephrin和ZO-1、间充质细胞标志性分子Ⅰ型胶原(collagenⅠ)和纤维连接蛋白(fibronectin,FN)的蛋白表达水平;最后,检测足细胞NLRP3炎症小体活化关键信号分子NLRP3,凋亡相关斑点样蛋白(apoptosis-associated speck-like protien,ASC)以及下游效应蛋白caspase-1、白介素(interleutin,IL)-1β、IL-18的蛋白表达水平。结果表明,对于体外培养的足细胞,HG能引起nephrin,ZO-1蛋白表达水平下调,collagenⅠ,FN蛋白表达水平上调,诱导其发生EMT;还能引起NLRP3,ASC以及caspase-1,IL-1β,IL-18蛋白表达水平上调,诱导其NLRP3炎症小体活化。此外,L-TP或H-TP与HG联合干预足细胞能恢复nephrin,ZO-1蛋白表达水平,抑制collagenⅠ,FN蛋白表达水平,改善EMT;还能下调NLRP3,ASC蛋白表达水平,抑制NLRP3炎症小体活化,并减少其下游效应分子caspase-1,IL-1β,IL-18蛋白表达水平。总之,HG在体外能激活NLRP3炎症小体而诱导足细胞EMT; TP在适当的剂量范围内能抑制NLRP3炎症小体活性而改善足细胞EMT,这可能是TP保护DKD足细胞炎症性损伤的关键分子机制之一。
The aim of this paper was to explore the effects of triptolide( TP),the effective component of Tripterygium wilfordii on improving podocyte epithelial-mesenchymal transition( EMT) induced by high glucose( HG),based on the regulative mechanisms of Nod-like receptor protein 3( NLRP 3) inflammasome in the kidney of diabetic kidney disease( DKD). The immortalized podocytes of mice in vitro were divided into the normal( N) group,the HG( HG) group,the low dose of TP( L-TP) group,the high dose of TP( HTP) group and the mannitol( MNT) group,and treated by the different measures,respectively. More specifically,the podocytes in each group were separately treated by D-glucose( DG,5 mmol·L~(-1)) or HG( 30 mmol·L~(-1)) or HG( 30 mmol·L~(-1)) + TP( 5 μg·L~(-1))or HG( 30 mmol·L~(-1)) + TP( 10 μg·L~(-1)) or DG( 5 mmol·L~(-1)) + MNT( 24. 5 mmol·L~(-1)). After the treatment of HG or TP at 24,48 and 72 h,firstly,the activation of podocyte proliferation was investigated. Secondly,the protein expression levels of the epithelial markers in podocytes such as nephrin and ZO-1,the mesenchymal markers such as collagen Ⅰ and fibronectin( FN) were detected,respectively. Finally,the protein expression levels of NLRP3 and apoptosis-associated speck-like protein( ASC) as the key signaling molecules of NLRP3 inflammasome activation,as well as the downstream effector proteins including caspase-1,interleutin( IL)-1β and IL-18 were examined,severally. The results indicated that,for the cultured podocytes in vitro,HG could cause the low protein expression levels of nephrin and ZO-1,induce the high protein expression levels of collagen Ⅰ and FN and trigger podocyte EMT. Also HG could cause the high protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 and induce NLRP3 inflammasome activation. On the other hand,the co-treatment of TP( L-TP or H-TP) and HG for podocytes could recover the protein expression levels of nephrin and ZO-1,inhibit the protein expression levels of collagen Ⅰ and FN and ameliorate podocyte EMT. Also the co-treatment of TP( L-TP or H-TP) and HG could down-regulate the protein expression levels of NLRP3 and ASC,inhibit NLRP3 inflammasome activation and reduce the protein expression levels of the downstream effector molecules including caspase-1,IL-1β and IL-18. On the whole,HG could activate NLRP3 inflammasome and induce podocyte EMT in vitro. TP at the appropriate dose range could inhibit NLRP3 inflammasome activation and ameliorate podocyte EMT,which may be one of the critical molecular mechanisms of TP protecting againstpodocyte inflammatory injury in DKD.

关键词(KeyWords): 糖尿病肾病;雷公藤甲素;足细胞;NLRP3炎症小体活化;上皮-间充质转分化
diabetic nephropathy;triptolide;podocyte;NLRP3 inflammasome activation;epithelial-mesenchymal transition

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(81573903,81774245);国家自然科学基金青年基金项目(81603675);; 江苏省自然科学基金青年基金项目(BK20161046);; 江苏省中医药科技发展计划项目(YB201937);; 南京市医学科技发展基金项目(QRX17042)

作者(Author): 吴薇;刘不悔;万毅刚;孙伟;刘莹露;王文文;房其军;涂玥;YEE Hong-yun;袁灿灿;万子玥;
WU Wei;LIU Bu-hui;WAN Yi-gang;SUN Wei;LIU Ying-lu;WANG Wen-wen;FANG Qi-jun;TU Yue;YEE Hong-yun;YUAN Can-can;WAN Zi-yue;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital Clinical College of Chinese Medicine and Western Medicine,Nanjing University of Chinese Medicine;Department of Nephrology,the Affiliated Hospital of Nanjing University of Chinese

Email:

DOI:

参考文献(References):

扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享