摘要
目的探讨SFRP1是否通过抑制Wnt/β-catenin通路促进心肌细胞增殖.方法选取c57BL/6雄性小鼠18只,出生后1d、7d及28d的c57BL/6小鼠各6只分为3组,分离和提取心肌组织,检测心肌组织中SFRP1的mRNA及蛋白表达水平.利用出生后1d的乳鼠心脏培养小鼠原代心肌细胞,予SFRP1-shRNA慢病毒转染细胞,免疫荧光检测心肌细胞增殖标志物Ki67和PH3表达变化,Westernblot检测胞浆β-catenin蛋白表达变化.结果SFRP1 mRNA和蛋白水平在小鼠出生后逐渐下降(P<0.05).转染SFRP1-shRNA慢病毒后,SFRP1蛋白表达水平均下降(P<0.05),其中shRNA-2干扰效果最好.与空载组相比,shRNA组细胞Ki67和PH3表达下降,同时胞浆β-catenin蛋白表达水平升高(P<0.05).结论SFRP1通过抑制Wnt/β-catenin信号通路促进心肌细胞增殖.
Objective To investigate whether SFRP1 promotes cardiomyocyte proliferation by inhibiting the Wnt/β-catenin pathway.Methods Six c57 BL/6 mice were selected which were 1/7/28-day old, respectively.The myocardial tissue was isolated and extracted, and the mRNA and protein expression levels of SFRP1 in myocardial tissue were detected.Primary mouse cardiomyocytes were cultured and identified from 1-day old mice.Cells were transfected with SFRP1-shRNA lentivirus.The expression proliferation markers of cardiomyocytes(Ki67,PH3) were detected by immunofluorescence, and the expression levels of β-catenin protein in cytoplasm was detected by WB.Results The mRNA and protein expression of SFRP1 decreased gradually after birth(P<0.05).The expression levels of SFRP1 protein was decreased after transfered with SFRP1-shRNA lentivirus, and the interference effect of shRNA-2 was the best.Compared with vector group, the expression of Ki67 and PH3 were decreased in shRNA group, and the expression of β-catenin protein in cytoplasm was increased(P<0.01).Conclusion SFRP1 promotes cardiomyocyte proliferation by inhibiting the Wnt/β-catenin pathway.
作者
傅明炜
王热华
林锋
郭延松
FU Mingwei;WANG Rehua;LIN Feng;GUO Yansong(Department of Cardiology,Fujian Provincial Hospital,Provincial Clinical College of Fujian Medical University,Fuzhou,Fujian 350001,China)
出处
《福建医药杂志》
CAS
2022年第6期117-120,共4页
Fujian Medical Journal
基金
福建省卫生健康科研人才培养项目(2019-1-8)。
