摘要
目的研究柚皮素预处理对大鼠缺血再灌注(I/R)心肌氧化损伤的作用及其分子机制。方法32只成年雄性SD大鼠分为假手术组(SHAM组)、I/R组、柚皮素组、柚皮素+LY294002组(NL组)。柚皮素组及NL组腹腔注射柚皮素(100 mg·kg^(-1)·d^(-1)),连续7 d后,NL组在结扎冠状动脉前30 min腹腔注射磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)通道抑制剂LY294002(0.3 mg/kg),除SHAM组,其余3组大鼠均进行冠状动脉结扎30 min再灌注120 min,造模成功后处死各组大鼠,苏木素-伊红(HE)染色观察心肌组织病理变化,TUNEL法检测心肌组织凋亡情况,使用试剂盒检测血清乳酸脱氢酶(LDH)、肌酸激酶(CK)、超氧化物歧化酶(SOD)、丙二醛(MDA)、活性氧(ROS)水平,Western blot测定心肌组织AKT、磷酸化蛋白激酶B(p-AKT)、醌氧化还原酶1(NQO1)、核因子E相关因子2(Nrf2)、血红素氧合酶1(HO-1)的表达水平,逆转录PCR(RT-PCR)测定Nrf2、HO-1转录水平。结果各组AKT表达水平比较,差异无统计学意义(P>0.05)。与SHAM组比较,I/R组LDH、CK、心肌细胞凋亡率、MDA、ROS水平升高,而SOD活性降低(P<0.05)。与I/R组比较,柚皮素组LDH、CK、心肌细胞凋亡率、MDA、ROS水平降低,而SOD活性、p-AKT、Nrf2、HO-1、NQO1表达水平升高(P<0.05)。与柚皮素组比较,NL组LDH、CK、心肌细胞凋亡率、MDA、ROS水平升高,而SOD活性、p-AKT、Nrf2、HO-1、NQO1表达水平降低(P<0.05)。结论柚皮素调节PI3K/AKT/Nrf2通路减轻心肌I/R氧化损伤。
Objective To investigate the protective effect and molecular mechanism of naringenin preconditioning on oxidative stress injury of myocardial ischemia reperfusion(I/R)in rats.Methods 32 adult male SD rats were randomly divided into the SHAM group,the I/R group,the naringenin group and the naringenin+LY294002 group(the NL group).The naringenin group and the NL group were intraperitoneally injected naringenin(100 mg·kg^(-1)·d^(-1))for 7 consecutive days,the NL group was intraperitoneally injected phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)channel inhibitor LY294002(0.3 mg/kg)30 minutes before coronary artery ligation.Except for the SHAM group,in the other three groups the rats were all subjected to coronary artery ligation for 30 minutes and reperfusion after 120 minutes,the rats of each group were sacrificed after successfully modeling.The pathological changes of the myocardial tissues were observed with HE staining,the apoptosis of myocardial tissues was detected by TUNEL,lactate dehydrogenase(LDH),creatine kinase(CK),superoxide dismutase(SOD),malondialdehyde(MDA),reactive oxygen species(ROS)contents were detected by kits.Western blot measures the protein levels of AKT,phosphorylated protein kinase B(p-AKT),quinone oxidoreductase 1(NQO1),nuclear factor E related factor 2(Nrf2),heme oxygenase 1(HO-1)in heart tissues.The transcription levels of Nrf2 and HO-1 were measured by reverse transcription PCR(RT-PCR)technology.Results There was no significant difference in the expression level of AKT in each group(P>0.05).Compared with the SHAM group,LDH,CK,myocardial cell apoptosis rate,MDA and ROS levels in the I/R group increased,while SOD activity decreased(P<0.05).Compared with the I/R group,the levels of LDH,CK,cardiomyocyte apoptosis,MDA and ROS in the naringenin group decreased,while SOD activity,p-AKT,Nrf2,HO-1 and NQO1 expression levels increased(P<0.05).Compared with the naringenin group,the levels of LDH,CK,cardiomyocyte apoptosis,MDA and ROS in the NL group increased,while SOD activity,p-AKT,Nrf2,HO-1 and NQO1 expression levels decreased(P<0.05).Conclusion Naringenin regulated PI3K/AKT/Nrf2 pathway to alleviate the oxidative of myocardial ischemia-reperfusion injury.
作者
王欢
李家富
兰卓
徐楠
程晓洪
冯健
WANG Huan;LI Jiafu;LAN Zhuo;XU Nan;CHENG Xiaohong;FENG Jian(Department of Cardiology,the Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan 646000,China)
出处
《重庆医学》
CAS
2021年第18期3066-3072,共7页
Chongqing medicine
基金
国家自然科学基金项目(31300946)
四川省2011项目-四川省心血管疾病防治协同创新中心基金项目(xtcx2019-14)
四川省泸州市-川医大联合课题[2015LZCYD-S03(7/7)]。
作者简介
王欢(1994—),在读硕士研究生,主要从事冠心病基础及临床治疗研究;通信作者:李家富,E-mail:ljf198@126.com。
