摘要
目的:探究神经纤毛蛋白-1(neuropilin-1)过表达对异丙肾上腺素(isoproterenol,ISO)诱导的心衰大鼠心脏功能及心肌组织损伤的保护作用。方法:构建neuropilin-1过表达的腺病毒载体(Ad-neuropilin-1),采用逆转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)结合Western blot检测过表达效率;选取60只SD大鼠,随机分为对照(control)组、ISO组、ISO+Ad(阴性对照)组和ISO+Ad-neuropilin-1组,除control组外各组采用皮下注射ISO制成心衰大鼠模型,检测各组大鼠心脏功能,采用RT-PCR和酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测各组大鼠心损标记物、炎性因子水平,HE染色、缺口末端标记法(TdT-medated dUTP nick end labeling,TUNEL)染色观察心肌细胞凋亡情况,Western blot检测增殖细胞相关抗原Ki-67和半胱天冬酶(cysteine aspartase-3,Cas3)的表达。结果:相比control组,ISO+Ad-neuropilin-1组neuropilin-1 mRNA和蛋白表达均升高(P<0.05);相比control组,ISO组大鼠心率(heart rate,HR)、左心室射血分数(left ventricular ejection fraction,LVEF)、左室短轴缩短率(left ventricular fraction shortening,LVFS)、neuropilin-1、Ki-67蛋白表达水平均明显降低,血清心肌肌钙蛋白(cardiac troponin,c TnI)、肌酸激酶(creatine kinase,CK)、肌酸激酶同工酶(creatine kinase-MB,CKMB)、一氧化氮合酶(inducible nitric oxide synthase,i NOS)和白细胞介素-10(interleukin-10,IL-10)mRNA水平、心肌细胞凋亡率、cleaved caspase-3/Cas3水平明显升高(P<0.05);相比ISO组,ISO+Ad-neuropilin-1组HR、LVFS、LVEF、neuropilin-1、Ki-67蛋白相对表达明显升高,cTnI、CK、CK-Mb、i NOS、IL-10 mRNA及外周血含量、心肌细胞凋亡率、cleaved caspase-3/Cas3水平均明显降低(P<0.05)。结论:neuropilin-1过表达可抑制ISO诱导的心衰大鼠心肌细胞凋亡及炎症反应,改善其心脏功能。
Objective:To explore the protective effect of neuropilin-1 overexpression on cardiac function and against myocardial tissue injury in rats with isoproterenol(ISO) induced heart failure(HF). Methods:The adenovirus vector of neuropilin-1 overexpression(Ad-neuropilin-1)was constructed. The overexpression efficiency was detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blot. Sixty SD rats were enrolled and randomly divided into control group,ISO group,ISO+Ad group(negative control group)and ISO+Ad-neuropilin-1 group. Except control group,subcutaneous injection of ISO was conducted to prepare HF rats models. The cardiac function in each group was detected. The levels of myocardial damage markers and inflammatory factors in each group were detected by RT-PCR and enzyme-linked immunosorbent assay(ELISA). The apoptosis of myocardial cells was observed by HE staining and TdT-medated dUTP nick end labeling(TUNEL) staining. The expression of proliferation cells related antigen Ki-67 and cysteine aspartase-3(Cas3) was detected by Western blot. Results:Compared with control group,neuropilin-1 m RNA and protein in ISO+Ad-neuropilin-1 group were increased(P <0.05). Compared with control group,heart rate(HR),left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),and expression levels of neuropilin-1 and Ki-67 proteins were significantly decreased in ISO group,while levels of serum cardiac troponin(cTnI),creatine kinase(CK),crea tine kinase-MB(CK-MB),inducible nitric oxide synthase(iNOS) and interleukin-10(IL-10) mRNA,apoptosis rate of myocardial cells and level of cleaved caspase-3/Cas3 were significantly increased(P <0.05). Compared with ISO group,HR,LVFS,LVEF,and relative expression of neuropilin-1 and Ki-67 proteins in ISO+Ad-neuropilin-1 group were significantly increased,while cTnI,CK,CK-Mb,iNOS,content of peripheral blood IL-10 mRNA,apoptosis rate of myocardial cells,and level of cleaved caspase-3/Cas3 were significantly decreased(P<0.05). Conclusion:Neuropilin-1
作者
蒋芳萍
赵玲
吴校伟
李韫
赵天华
谢金洲
Jiang Fangping;Zhao Ling;Wu Xiaowei;Li Yun;Zhao Tianhua;Xie Jinzhou(Department of Cardiovascular Medicine,The 363th Hospital;Department of Cardiovascular Medicine,Chengdu Sixth People’s Hospital)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2021年第12期1416-1420,共5页
Journal of Chongqing Medical University
基金
四川省卫计委科研资助项目(编号:17PJ1183)。
