脂联素对血管紧张素Ⅱ诱导的小鼠心脏成纤维细胞增殖及ERK1/2信号的影响  

Effects of adiponectin on angiotensin-Ⅱ-induced mouse cardiac fibroblast proliferation and ERK1/2 signaling

作　　者：刘立波 刘志坚 陈丽娜 刘晓红 李秀昌 LIU Libo;LIU Zhijian;CHEN Lina;LIU Xiaohong;LI Xiuchang(Department of Cardiology,The Second Affiliated Hospital of Shandong First Medical University,Taian 271000,China;不详)

机构地区：[1]山东第一医科大学第二附属医院心脏中心,山东泰安271000 [2]山东第一医科大学研究生部

出　　处：《山东医药》2021年第31期47-50,共4页Shandong Medical Journal

摘　　要：目的探讨脂联素(APN)对血管紧张素Ⅱ(AngⅡ)诱导的小鼠心脏成纤维细胞(CF)增殖及ERK1/2信号的影响。方法取日龄1~3 d的C57乳鼠心室肌组织,采用冷胰酶联合Ⅰ型胶原酶消化法分离细胞,贴壁差速法获得纯化小鼠CF,用形态学及免疫细胞化学技术进行鉴定。取2~4代CF随机分为5组,对照组用无血清的DMEM培养基培养;AngⅡ组用含1×10^(-7)mol/L AngⅡ的DMEM培养基培养;APN低、中、高剂量组先分别用1、5、25μg/mL APN干预1 h,然后加含1×10^(-7)mol/L AngⅡ的DMEM培养基培养。均培养24 h后收集细胞。用MTT法检测CF增殖能力(OD值);用Western blotting法检测CF内细胞外调节蛋白激酶1/2(ERK1/2)、磷酸化的细胞外调节蛋白激酶1/2(p-ERK1/2)蛋白表达,并计算p-ERK/ERK。结果成功分离小鼠心脏CF,经鉴定细胞纯度接近100%。与对照组比较,AngⅡ组OD值高(P<0.05);与AngⅡ组比较,各APN组OD值低(P均<0.05);各APN组OD值比较差异无统计学意义(P均>0.05)。各组ERK1/2蛋白相对表达量比较差异无统计学意义(P均>0.05);与对照组比较,AngⅡ组p-ERK蛋白相对表达量、p-ERK/ERK高(P均<0.05);与AngⅡ组比较,APN高剂量组p-ERK蛋白相对表达量、p-ERK/ERK低(P均<0.05)。结论脂联素可抑制AngⅡ诱导的小鼠心脏CF增殖及ERK1/2信号激活。Objective To observe the effects of adiponectin(APN) on angiotensin-Ⅱ(Ang Ⅱ)-induced cardiac fibroblasts(CFs) proliferation and extracellular signal-regulated kinase 1/2(ERK1/2) activation.Methods Cardiac ventricles were separated from 1 to 3-day-old C57 mice.Pure primary mouse cardiac fibroblasts were obtained after cardiomyocytes were separated by trypsin-collagenase type I digestion and differential adhesion.Morphological and immunocytochemical technique was used to identify the CFs.CFs of two to four generations were randomly divided into five groups.CFs in the control group were cultured with serum-free DMEM medium.CFs in the Ang Ⅱ group were cultured with DMEM medium containing 10^(-7) mol/L Ang Ⅱ.CFs in the low-dose,medium-dose and high-dose APN groups were treated with 1,5 and 25 μg/mL APN for 1 h,respectively,and then were cultured with DMEM medium containing 10^(-7) mol/L Ang Ⅱ.The cells were collected after 24 h culture.The proliferation ability(OD) of CFs was measured by MTT,the expression of ERK1/2 and phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2) in CFs was detected by Western blotting,and the p-ERK/ERK ratio was calculated.Results CFs were successfully isolated from mouse heart,and the cell purity was close to 100% by immunofluorescence combined with morphological identification.Compared with the control group,the OD value of the Ang Ⅱ group increased(P<0.05);compared with the Ang Ⅱ group,OD values of the APN groups were lower(all P<0.05);there was no statistically significant difference in OD value among APN groups(P>0.05).There was no statistically significant difference in the relative expression of ERK1/2 protein among all groups(P>0.05).Compared with the control group,the relative expression of p-ERK protein and the ratio of p-ERK/ERK were higher in the Ang Ⅱ group(all P<0.05).Compared with the Ang Ⅱ group,p-ERK protein relative expression level and the ratio of p-ERK/ERK were lower in the high-dose APN group(all P<0.05).Conclusion Adiponectin inhibits t

关 键 词：脂联素 血管紧张素Ⅱ 心脏成纤维细胞 细胞增殖 细胞外调节蛋白激酶1/2 

分 类 号：R542.2[医药卫生—心血管疾病]