文章摘要
虞天一,赵晨卉,何风霞,刘玉,宫雅娟,赵聃,邱文,张婧,王迎伟.Sublytic C5b⁃9刺激上调KLF4促进肾小球系膜细胞生成IL⁃23的作用[J].南京医科大学学报,2018,(10):1331~1336,1387
Sublytic C5b⁃9刺激上调KLF4促进肾小球系膜细胞生成IL⁃23的作用
Sublytic C5b ⁃ 9 induces IL ⁃ 23 production of rat glomerular mesangial cells via up ⁃regulated KLF4 expression
投稿时间:2018-02-24  
DOI:10.7655/NYDXBNS20181001
中文关键词: Sublytic C5b⁃9  肾小球系膜细胞(GMC)  KLF4  IL⁃23
英文关键词: sublytic C5b⁃9  glomerular mesangial cells(GMC)  KLF4  IL⁃23
基金项目:国家自然科学基金(31500701,81471626);江苏省自然科学青年基金(BK20140910)
作者单位
虞天一 南京医科大学免疫学系江苏南京211166
东南大学附属中大医院妇产科江苏南京210009 
赵晨卉 南京医科大学第一附属医院肿瘤科江苏南京210029 
何风霞 南京医科大学免疫学系江苏南京211166
南京医科大学第二附属医院病理科江苏南京210003 
刘玉 南京医科大学免疫学系江苏南京211166 
宫雅娟 南京医科大学免疫学系江苏南京211166 
赵聃 南京医科大学免疫学系江苏南京211166 
邱文 南京医科大学免疫学系江苏南京211166 
张婧 南京医科大学免疫学系江苏南京211166 
王迎伟 南京医科大学免疫学系江苏南京211166 
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中文摘要:
      目的:研究转录因子KLF4调控sublytic C5b?9刺激肾小球系膜细胞(glomerular mesangial cell,GMC)诱导促炎因子白介素?23(IL?23)生成的作用。方法:构建KLF4的短发夹状小干扰RNA(shKLF4)及过表达质粒(pIRES2?KLF4)。将pIRES2?KLF4或shKLF4 转染GMC后再行sublytic C5b?9刺激,用qPCR、Western blot和ELISA法检查沉默或过表达KLF4基因后对GMC产生IL?23的影响。此外,构建IL?23基因近端启动子全长质粒,行荧光素酶报告基因实验测定沉默或过表达KLF4基因后对IL?23启动子活性的影响。结果:①Sublytic C5b?9刺激GMC后能明显促进IL?23的生成,而沉默KLF4基因后,由sublytic C5b?9诱导GMC产生的IL?23明显减少,但过表达KLF4后IL?23的水平则显著增加。②Sublytic C5b?9刺激GMC能显著上调IL?23的启动子活性,而沉默KLF4基因后由sublytic C5b?9诱导的IL?23启动子活性明显降低,但过表达KLF4后又能显著升高IL?23的启动子活性。结论:Sublytic C5b?9刺激诱导IL?23的生成可通过其刺激上调转录因子KLF4的表达而实现。
英文摘要:
      Objective:To explore the regulatory role of KLF4,as a transcription factor,in the production of pro?inflammation cytokine IL?23 gene in rat glomerular mesangial cells(GMC)induced by sublytic C5b?9. Methods:The plasmids of KLF4 short hairpin RNA(shKLF4)and KLF4 overexpression(pIRES2?KLF4)were first generated and transfected rat GMC respectively,then the cells were treated with or without sublytic C5b?9. The roles of silencing KLF4 gene in IL?23 synthesis induced by sublytic C5b?9 or overexpressing KLF4 gene in IL?23 production were detected by qPCR,Western blot and ELISA. Moreover,the plasmid of IL?23 promoter(full?length,FL)was also constructed. After co?transfection,the activity of IL?23 promoter with or without sublytic C5b?9 after KLF4 gene knockdown or overexpression was measured by luciferase reporter assay. Results:①Sublytic C5b?9 could obviously elevate IL?23 production in rat GMC,and after KLF4 gene knochdown with shKLF4,the increase of IL?23 expression upon sublytic C5b?9 was remarkably diminished,but KLF4 overexpression in the GMC could markedly elevate IL?23 synthesis. ②The IL?23 promoter activity incubated with sublytic C5b?9 was significantly up?regulated. Meanwhile,after KLF4 gene knockdown,the IL?23 promoter activity in the GMC stimulated by sublytic C5b?9 was obviously declined,but KLF4 overexpression greatly increased IL?23 promoter activivy. Conclusion:Sublytic C5b?9 could induce IL?23 production of GMCs through the increase of KLF4 expression.
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