Effects of E3 ubiquitin-protein ligase c-casitas B-lineage lymphoma on phenotypic transformation of rat primary cardiac fibroblasts after angiotensin Ⅱ stimulation
LIU Mei-li
LIU Dan
CHENG Xiao-li
XING Rui-nan
TIAN Xiao-xiang
YAN Cheng-hui
Abstract：Objective To investigate the effect of E3 ubiquitin ligase c-casitas B-lineage lymphoma (c-CBL) on angiotensinⅡ (AngⅡ) induced mice after the original generation of myocardial fibroblasts phenotypic transition.Methods Cells can be divided into the control group (not stimulated), AngⅡstimulated group (different concentrations of AngⅡstimulates cell for 24 hours), fluorescence quantitative (polymerase chain reacion, PCR) and Western blot test c-CBL andαsmooth muscle actin (αSMA) expression.Application of small interfering RNA transfection technique to establish a lower expression c-CBL (si-CBL) cells, the cells into si-control group (small interference RNA contrast transfection for 24 hours), si-CBL group (c-CBL small interference RNA transfection for 24 hours), si-control+AngⅡgroup (small interference RNA controls transfection after 24 hours to give AngⅡstimulus for 24 hours) and siCBL+AngⅡgroup (c-CBL small interference RNA transfection after 24 hours to give AngⅡstimulus for 24 hours). Application of fluorescence quantitative PCR and Western blot test each cell c-CBL, αSMA and collagen 1 expression, application of immunofluorescence staining detection collagen 1 groups of cells.ResultsTheαSMA、c-CBL gene mRNA and protein AngⅡgroup were significantly higher than the control group (P＜0.05). The mRNA and protein expression levels of c-CBL gene in the si-CBL group were significantly lower than those in the si-control group (P＜0.05). The mRNA and protein expression levels of c-CBL gene in the si-CBL group were significantly lower than those in the si-control group (P＜0.05). AngⅡstimulation, si-control+AngⅡgroup and si-CBL+AngⅡαSMA gene mRNA and protein expression of si-control group and si-CBL group was obviously increased;αSMA gene mRNA and protein of si-CBL+AngⅡgroup were significantly lower than those of si-control+AngⅡgroup (P＜0.05). After AngⅡstimulation, αSMA gene mRNA and protein expression of si-control+AngⅡgroup and si-CBL+AngⅡgroup were increased more obviously than those in the si-control group and si-CBL group;theαSMA gene mRNA and protein expression of si-control+AngⅡgroup and si-CBL+AngⅡof si-control group and si-CBL group was obviously increased;αSMA gene mRNA and protein of si-CBL+AngⅡgroup were significantly lower than those of si-control+AngⅡgroup (P＜0.05). ConclusionThe c-CBL could affect the phenotypic transformation of rat primary cardiac fibroblasts after AngⅡstimulation.
Keywords：E3 ubiquitin-protein ligasec-casitas B-lineage lymphomaAngiotensin ⅡCardiac fibroblastsPhenotypic transformation
Publication Date：2019-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 5-9 )