摘要
目的观察核因子E2相关因子2(nuclear factor-erythroid 2-related factor 2,Nrf2)在白藜芦醇(resveratrol,Res)预处理减轻糖尿病大鼠心肌缺血/再灌注损伤(myocardial ischemia/reperfusion injury,MI/RI)中的作用。方法清洁级健康成年雄性SD大鼠,体重200~220 g,4~5月龄,采用先喂养高脂饲料4周后,一次性腹腔注射链脲佐菌(streptozotocin,STZ)30 mg/kg的方法制备糖尿病大鼠模型。取糖尿病模型制备成功的大鼠30只,采用随机数字表法分为3组(每组10只):假手术组(Sham组)、心肌缺血/再灌注组(MI/R组)和心肌缺血/再灌注+Res组(MI/R+Res组)。采用结扎冠状动脉左前降支30 min,恢复灌注120 min的方法制备大鼠MI/RI模型。MI/R+Res组于MI/RI模型建立前7 d腹腔注射Res 15 mg/kg,1次/d,连续7 d。Sham组与MI/R组腹腔注射等容量二甲基亚砜(dimethyl sulfoxide,DMSO)与PBS的混合液。B超测量左心室射血分数(left ventricular ejection fraction,LVEF)以及左室短轴缩短率(fractional shortening,FS)。于再灌注120 min时处死5只大鼠,用2,3,5氯化三苯基四氮唑(2,3,5-Triphenyl-2H-tetrazolium chloride,TTC)染色法测定心肌梗死面积;另再处死5只大鼠,经腹主动脉取血测心肌损伤标志物乳酸脱氢酶(lactate dehydrogenase,LDH)和肌酸激酶同工酶(creatine kinase isoenzyme,CK-MB)的浓度,并取心肌组织,采用ELISA法检测心肌组织的超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)及活性氧自由基(reactive oxygen species,ROS)的含量,免疫组织化学法观察Nrf2的含量,Western blot法测定心肌组织Nrf2、细胞核-核因子E2相关因子2(n-nuclear factor-erythroid 2-related factor 2,n-Nrf2)和血红素加氧酶-1(heme oxygenase-1,HO-1)的表达。结果与Sham组比较,MI/R组与MI/R+Res组的LDH和CK-MB浓度、ROS及MDA含量均显著增高,LVEF和FS、SOD活性显著降低,心肌梗死面积增加,Nrf2、n-Nrf2与HO-1蛋白表达下调(P均<0.05);与MI/R组比较,MI/R+Res组的LDH和CK-MB浓度、ROS及MDA含量均显著降低,LVEF和FS、SOD活性显著增加,心肌梗死面积减少,Nrf2、n-Nrf2与HO-1蛋白表达上调(P均<0.05)。结论RES预处理减轻糖尿病大鼠MI/RI的作用可能与激活Nrf2有关。
Objective To observe the effect of nuclear factor-erythroid 2-related factor 2(Nrf2)in diabetic rats preconditioning with resveratrol(Res)against myocardial ischemia/reperfusion injury(MI/RI).Methods Healthy adult male SD rats weighing 200-220 g and aged from 4 to 5 months were fed with high-fat diet for 4 weeks and then intraperitoneally injected with streptozotocin(STZ)30 mg/kg to establish a diabetic rat model.Thirty successfully modeled rats were divided into three groups according to a random number table(n=10):a sham operation group(a Sham group),a myocardial ischemia/reperfusion group(an MI/R group)and a myocardial ischemia/reperfusion+resveratrol group(an MI/R+Res group).A rat model of MI/RI was established by ligation of the left anterior descending coronary artery for 30 min before reperfusion for 120 min.In the MI/R+Res group,15 mg/kg Res was intraperitoneally injected 7 d before establishment of the MI/RI model,once a day for 7 consecutive days.The Sham and MI/R groups were intraperitoneally injected with an equal volume of dimethyl sulfoxide(DMSO)and PBS mixture.Left ventricular ejection fraction(LVEF)and left ventricular short axis shortening(FS)were measured by B-mode ultrasonography.Five rats were sacrificed 120 min after reperfusion.Myocardial infarction size was measured by 2,3,5-triphenyl-2H-tetrazolium chloride(TTC)staining.Another five rats were sacrificed to collect blood samples from the abdominal aorta for measurement of lactate dehydrogenase(LDH),a marker of myocardial injury,and creatine kinase isoenzyme(CK-MB).Their myocardial tissues were collected to determine the levels of superoxide dismutase(SOD),malondialdehyde(MDA)and reactive oxygen species(ROS)by enzyme-linked immunosorbent assay(ELISA).The contents of Nrf2 were observed by immunohistochemistry.The expression of Nrf2,n-Nrf2 and heme oxygenase-1(HO-1)were determined by Western blot.Results Compared with those in the Sham group,LDH,CK-MB concentration,ROS and MDA content in the MI/R and MI/R+Res groups significantly increased while LVEF,FS,and SOD activity in the MI/R and MI/R+Res groups significantly decreased.Meanwhile,the myocardial infarction sizes in the MI/R and MI/R+Res groups were increased,and the expression of Nrf2,n-Nrf2 and HO-1 protein was down-regulated(all P<0.05).Compared with those in the MI/R group,LDH,CK-MB concentration,ROS and MDA content significantly decreased in the MI/R+Res group,while LVEF,FS and SOD activity significantly increased in the MI/R+Res group.Also,in the MI/R+Res group,the myocardial infarction size decreased while the expression of Nrf2,n-Nrf2 and HO-1 protein increased(all P<0.05).Conclusions Resveratrol preconditioning relieves myocardial injury in diabetic rats with ischemia/reperfusion,which may attribute to activation of Nrf2.
作者
徐桂萍
胡东
王晓丽
赵萱付
鹃何虹
Xu Guiping;Hu Dong;Wang Xiaoli;Zhao Xuan;Fu Juan;He Hong(Department of Anesthesiology,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)
出处
《国际麻醉学与复苏杂志》
CAS
2020年第1期11-17,共7页
International Journal of Anesthesiology and Resuscitation
基金
新疆维吾尔自治区自然科学基金(2018D01C110)。
作者简介
通信作者:徐桂萍,Email:XGPSYL@126.com。
