摘要
目的研究非受体酪氨酸激酶肉瘤病毒蛋白(Src)在异丙肾上腺素(ISO)诱导的小鼠脂肪分解中的作用。方法将10周龄的雄性C57BL/6野生型小鼠分成3组:对照组、ISO组和ISO+吡唑并嘧啶类化合物(PP1)组。检测各组小鼠的附睾脂肪重量,苏木精和伊红(HE)染色观察脂肪细胞大小;实时荧光定量聚合酶链反应(qPCR)技术检测附睾脂肪组织中脂肪分解基因--脂肪组织三酰甘油脂肪酶(ATGL)及脂肪合成基因--过氧化物酶体增殖物激活受体γ(PPARγ)和脂蛋白脂肪酶(LPL)的mRNA表达。3T3-L1脂肪细胞处理分为3组:对照组、ISO组和ISO+PP1组。蛋白免疫印迹法检测不同处理条件下Src的磷酸化变化。结果动物实验结果显示,与对照组相比,ISO组小鼠的附睾脂肪重量减少,脂肪细胞大小显著降低(均为P<0.001);qPCR结果显示,与对照组相比,ISO组脂肪分解基因ATGL的mRNA表达显著增加(P<0.05),脂肪合成基因PPARγ和LPL的mRNA表达显著减少(均为P<0.01)。此外,在动物实验中我们证实了Src在脂肪分解中的作用,结果显示与ISO组比较,ISO+PP1组小鼠的附睾脂肪重量(P<0.05)及脂肪细胞大小(P<0.001)均明显增加;qPCR结果显示,与ISO组相比,ISO+PP1组脂肪分解基因ATGL的mRNA表达下降(P<0.01),脂肪合成基因PPARγ(P<0.01)和LPL的mRNA表达上升。细胞实验结果表明,ISO能显著增加Src的磷酸化(P<0.05),且可被PP1有效抑制(P<0.01)。结论在实验动物体内,ISO诱导脂肪分解依赖Src激酶活性。
Objective To study the role of non-receptor tyrosine kinase sarcoma virus protein(Src)in isoproterenol(ISO)-induced lipolysis in mice.Methods Male C57BL/6 wild-type mice aged 10 weeks were divided into 3 groups:control group,ISO group and ISO+pyrazolopyrimidine(PP1)group.Detect the weight of epididymal fat in each group of mice,and observe the size of fat cells by hematoxylin and eosin(HE)staining;Quantitative real-time polymerase chain reaction(qPCR)technology detect lipolytic genes in epididymal adipose tissue--adipose tissue triacylglycerol lipase(ATGL)and fat synthesis genes--peroxisome proliferator activation receptorγ(PPARγ)and lipoprotein lipase(LPL)mRNA expression.The 3T3-L1 adipocyte treatment was divided into 3 groups:control group,ISO group and ISO+PP1 group.Western blot was used to detect the phosphorylation changes of Src under different treatment conditions.Results The results of animal experiments showed that compared with the control group,the weight of epididymal fat in the ISO group mice was reduced,and the size of fat cells was significantly reduced(both P<0.001);the qPCR results showed that compared with the control group,the mRNA expression of the lipolytic gene ATGL in the ISO group increased significantly(P<0.05),and the mRNA expression of the fat synthesis genes PPARγand LPL decreased significantly(both P<0.01).In addition,in animal experiments,we confirmed the role of Src in lipolysis.The results showed that compared with the ISO group,the weight of epididymal fat(P<0.05)and the size of fat cells(P<0.001)of the ISO+PP1 group mice were both obviously increased.The qPCR results showed that compared with the ISO group,the mRNA expression of the lipolytic gene ATGL in the ISO+PP1 group decreased(P<0.01),and the mRNA expression of the fat synthesis genes PPARγ(P<0.01)and LPL increased.Cell experiment results showed that ISO can significantly increase the phosphorylation of Src(P<0.05),and it can be effectively inhibited by PP1(P<0.01).Conclusions In experimental animals,ISO-induced lipolys
作者
程扣辉
姜允奇
王文景
李文奇
李子健
马鑫
冯磊
Cheng Kouhui;Jiang Yunqi;Wang Wenjing;Li Wenqi;Li Zijian;Ma Xin;Feng Lei(School of Pharmacy and Wuxi School of Medicine,Jiangnan University,Wuxi 214122,China;Department of Cardiology and Institute of Vascular Medicine,Peking University Third Hospital,Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides,Ministry of He,Key Laboratory of Molecular Cardiovascular Science,Ministry of Education,Beijing Key Laboratory of Cardiovascular Receptors Research,Beijing 100191,China;Wuxi School of Medicine,Jiangnan University,Wuxi 214122,China)
出处
《中国心血管杂志》
2021年第6期569-574,共6页
Chinese Journal of Cardiovascular Medicine
基金
国家自然科学基金(91939301、81820108031)。
