Intervention of siRNA to the effect of gelatinase on the expression of ICAM-1 in human renal tubular epithelial cells
- VernacularTitle:siRNA干扰明胶酶对人肾小管细胞ICAM-1表达的影响
- Author:
Yan MEI
;
Quan HONG
;
Xueguang ZHANG
- Publication Type:Journal Article
- Keywords:
matrix metalloproteinases;
intercellular adhesion molecule-1;
RNA interference
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To inhibit the expression of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9(MMP-9) in human kidney tubular epithelial cell(HKC) with specific small interference RNA(siRNA),then to observe whether the expression of intercellular adhesion molecule-1(ICAM-1) on HKC could be influenced.Methods HKCs were transfected with MMP-2,MMP-9 siRNA or irrespective siRNA respectively,untransfected HKC was considered as blank control.Four groups of HKCs were stimulated with 100nmol/L phorbol myristate acetate(PMA) for 18h.Total RNA or protein of cells were extracted with Trizol and RIPA cell lysate respectively,the mRNA expression levels of MMP-2,MMP-9 and ICAM-1 were detected by real time PCR,the protein expression levels of above indexes were detected by western blotting and immunofluorescence combined with confocal laser microscopy.Results The specific siRNA could inhibit the expression of MMP-2 and MMP-9 in HKCs effectively.Immunofluorescence combined with confocal assay demonstrated that while HKCs was stimulated with PMA,the expression of ICAM-1 was up-regulated,which was significantly up-regulated in the group transfected with MMP-9 siRNA(P0.05).Conclusions The inhibitory expression of MMP-9 of HKC may up-regulate the expression of ICAM-1,implying that,besides taking part in the degradation of extracellular matrix,MMP-9 may throw an influence on the proceeding of renal fibrosis by inflammatory pathway.
