摘要
基于丝裂原活化蛋白激酶P38(P38 MAPK)/c-Jun氨基末端激酶(JNK)信号通路探讨二甲双胍对脓毒性心肌病的保护机制。本文为动物实验研究设计,于2023年1—12月在中南大学湘雅医院完成。采用随机数字表法将48只8周龄雌性C57BL/6小鼠分为4组:A组(对照组)、B组(模型组)、C组(模型+盐酸曲美他嗪)、D组(模型+二甲双胍组),每组12只。B、C、D组均以LPS(15 mg/kg)腹腔注射构建脓毒性心肌病小鼠模型,建模24 h后,A组、B组予以腹腔注射等量生理盐水,C组予以20 mg/kg盐酸曲美他嗪灌胃,D组予以腹腔注射二甲双胍200 mg/kg,均连续干预14 d。采用超声成像系统检测心功能,采用TUNEL法检测心肌组织细胞凋亡率;采用实时荧光定量聚合酶链反应(RT-qPCR)检测小鼠心肌组织中P38 MAPK/JNK信号通路JNK、P38 MAPK mRNA水平;采用酶联免疫吸附试验(ELISA)法测定各组小鼠血浆肌酸激酶同工酶(CK-MB)、脑利钠肽(BNP)、肿瘤坏死因子α(TNF-α)、白介素6(IL-6)含量;采用蛋白质印迹(Western blot)检测心肌组织中蛋白激酶C Eplison亚型(PKCε)、小窝蛋白-3(Cav-3)的蛋白表达。结果显示,与A组左心室射血分数(LVEF)(79.51±6.62)%、左室短轴缩短率(FS)(45.66±4.13)、细胞凋亡率(4.34±0.36)%、JNK(0.96±0.06)、P38 MAPK(1.01±0.03)、CK-MB(2.37±0.13)μg/L、BNP(21.36±3.47)ng/L、TNF-α(176.22±19.24)ng/L、IL-6(35.43±3.84)ng/L、PKCε表达(1.98±0.26)、Cav-3表达(1.04±0.03)相比,B、C、D组细胞凋亡率(28.22±4.49、22.45±3.69、15.88±3.27)%、JNK(1.68±0.11、1.32±0.18、1.13±0.14)、P38 MAPK(2.47±0.71、1.77±0.35、1.49±0.05)、CK-MB(16.55±2.16、12.63±1.98、5.27±0.61)μg/L、BNP(48.92±5.67、33.78±4.11、27.55±3.84)ng/L、TNF-α(463.71±24.81、335.71±36.71、214.78±22.53)ng/L、IL-6(78.57±6.36、63.71±5.66、52.47±5.47)ng/L表达均升高,而LVEF(49.38±5.27、55.47±5.03、62.26±5.14)%、FS(24.36±2.17、30.43±3.29、33.57±2.72)%、PKCε表达(1.33±0.21、1.54±0.23、1
Exploring the protective mechanism of metformin against septic cardiomyopathy based on the mitogen-activated protein kinase P38(P38 MAPK)/c-Jun amino-terminal kinase(JNK)signaling pathway.This paper is an experimental animal study design,which was completed from January to December 2023 at the Xiangya Hospital,Central South University.Forty-eight 8-week-old female C57BL/6 mice were divided into four groups:group A(control group),group B(model group),group C(model+trimetazidine hydrochloride),and group D(model+metformin group),with 12 mice in each group,by using a randomized numeric table method.Groups B,C,and D were injected intraperitoneally with LPS(15 mg/kg)to construct a septic cardiomyopathy mouse model.24 h after modeling,Groups A and B were injected intraperitoneally with an equal amount of saline,Group C was given 20 mg/kg trimetazidine hydrochloride by gavage,and Group D was injected with metformin 200 mg/kg intraperitoneally,and all of them were subjected to consecutive interventions for 14 d.The results were summarized in the following table.Ultrasound imaging system was used to detect cardiac function,and TUNEL method was used to detect apoptosis rate of myocardial tissues;real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the levels of mRNA of JNK,P38 MAPK of P38 MAPK signaling pathway in the myocardial tissues of mice;Plasma creatine kinase isoenzyme(CK-MB),brain natriuretic peptide(BNP),tumor necrosis factor alpha(TNF-α),and interleukin 6(IL-6)levels were measured by enzyme-linked immunosorbent assay(ELISA)in all groups of mice;and protein kinase C,and protein kinase C levels were measured by protein blotting in cardiac muscle tissue.Eplison isoform(PKCε),and Cavity protein-3(Cav-3)protein expression in myocardial tissues.The results showed that compared with group A,left ventricular ejection fraction(LVEF)(79.51±6.62)%,left ventricular short-axis shortening(FS)(45.66±4.13),apoptosis rate(4.34±0.36)%,JNK(0.96±0.06),P38 MAPK(1.01±0.03),CK-MB(2.37±0.13)μ
作者
李莉
廖杨
刘志勇
Li Li;Liao Yang;Liu Zhiyong(Department of Critical Care Medicine,Xiangya Hospital,Central South University,National Clinical Research Center for Geriatric Disorders(Xiangya Hospital),Changsha 410008,China)
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2024年第10期1567-1572,共6页
Chinese Journal of Preventive Medicine
基金
湖南省自然科学基金青年项目(2021JJ41001)。
