茶黄素对新生大鼠心肌缺血保护作用的机制研究  

Protective Effect of Theaflavin on Myocardial Ischemia in Neonatal Rats

作　　者：韩旭[1] 朱萍[1] 柳旎 吴宗跃[1] 杨静[1] HAN Xu;ZHU Ping;LIU Ni;WU Zongyue;YANG Jing(Nanyang Central Hospital,Nanyang 473001,Henan,China)

机构地区：[1]南阳市中心医院,河南南阳473001

出　　处：《中西医结合心脑血管病杂志》2023年第19期3521-3526,3539,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的:探讨茶黄素对新生大鼠心肌缺血保护作用的机制研究。方法:7日龄无特定病原体(SPF)新生Sprague-Dawley(SD)大鼠25只随机分为对照组、异丙肾上腺素(ISO)诱导组、茶黄素12.5 mg/kg组、茶黄素25.0 mg/kg组和茶黄素50.0 mg/kg组,每组5只。茶黄素12.5 mg/kg组、茶黄素25.0mg/kg组和茶黄素50.0 mg/kg组分别腹腔注射茶黄素12.5、25.0、50.0 mg/kg预处理7 d,每日1次。除对照组外,其余各组新生大鼠在预处理最后2 d皮下注射ISO 85 mg/kg构建心肌缺血模型。心脏超声检测心率(HR)、左室壁相对厚度(LVWT)、左室收缩末期容积(LVESV)和左室射血分数(LVEF);苏木精-伊红(HE)染色检测心脏组织损伤程度;酶联免疫吸附试验(ELISA)检测肌钙蛋白I(cTnI)、肌酸激酶同工酶(CK-MB)、肌红蛋白(Mb)、丙二醛(MDA)、超氧化物歧化酶(SOD)和髓过氧化物酶(MPO)的蛋白相对表达量;免疫组化检测Caspase-3阳性表达量;蛋白免疫印迹法(Western Blot)检测Cleaved Caspase-9、Bcl-2、Bax、鼠双微体2基因(MDM2)、p53、磷酸化的核因子E2相关因子2(p-Nrf2)、血红素加氧酶1(HO-1)和NADPH奎宁氧化还原酶1(NQO1)的蛋白相对表达量。结果:HE染色结果显示,与对照组比较,ISO诱导组大鼠心肌间隙增宽,心肌细胞严重肿胀,附着炎性细胞增加,结构变得模糊;在茶黄素治疗后,与ISO诱导组比较,以上现象均得到缓解。与对照组比较,ISO诱导组HR、LVWT和LVEF明显降低,LVESV明显升高(P<0.05或P<0.01);cTnI、CK-MB、Mb、MDA和MPO明显升高,SOD明显降低(P<0.05或P<0.01);Caspase-3阳性表达量、Cleaved Caspase-9/Caspase-9和p53的蛋白相对表达量明显上调,Bcl-2/Bax、MDM2、p-Nrf2/Nrf2、HO-1和NQO1的蛋白相对表达量明显下调(P<0.05)。与ISO诱导组比较,茶黄素25.0 mg/kg组、茶黄素50.0 mg/kg组HR、LVWT和LVEF明显升高,LVESV明显降低(P<0.05或P<0.01);cTnI、CK-MB、Mb、MDA和MPO明显降低,SOD明显升高(P<0.05或P<0.01);Caspase-3阳性表达量、CleavObjective:To explore the protective effect of theaflavin on myocardial ischemia in neonatal rats.Methods:Twenty-five 7-day-old Sprague-Dawley(SD)rats free of specific pathogens(SPF)were randomly divided into control group,isoproterenol(ISO)-inducted group,theaflavin 12.5 mg/kg group,theaflavin 25.0 mg/kg group and theaflavin 50.0 mg/kg group,with 5 in each group.Theaflavin 12.5 mg/kg group,theaflavin 25.0 mg/kg group and theaflavin 50.0 mg/kg group,were pretreated with intraperitoneal injection of theaflavin 12.5,25.0 and 50.0 mg/kg,once a day,for 7 days,respectively.The myocardial ischemia model was established by subcutaneous injection of ISO 85 mg/kg at the last 2 days of pretreatment.Heart rate(HR),left ventricular wall relative thickness(LVWT),left ventricular end-systolic volume(LVESV),and left ventricular ejection fraction(LVEF)were measured by echocardiography.Hematoxylin-eosin(HE)staining was used to detect the degree of cardiac tissue injury.The relative expression levels of troponin I(cTnI),creatine kinase isoenzyme(CK-MB),myoglobin(Mb),malonaldehyde(MDA),superoxide dismutase(SOD),and myeloperoxidase(MPO)were detected by enzyme-linked immunosorbent assay(ELISA).The positive expression of Caspase-3 was detected by immunohistochemistry.The relative protein expression levels of Cleaved Caspase-9,Bcl-2,Bax,MDM2,p53,phosphorylated nuclear factor E2-related factor 2(p-Nrf2),heme oxygenase-1(HO-1),and NADPH quinine oxidoreductase 1(NQO1)were detected by Western Blot.Results:HE staining results showed that compared with the control group,the myocardial space in the ISO-induced group widened,myocardial cells severely swollen,the attached inflammatory cells increased,and the structure became blurred.After theaflavin treatment,the above phenomena alleviated compared with ISO-induced group.Compared with the control group,HR,LVWT,and LVEF in ISO-induced group significantly decreased,while LVESV significantly increased(P<0.05 or P<0.01);cTnI,CK-MB,Mb,MDA,and MPO significantly increased,while SOD significantly decrea

关 键 词：心肌缺血 茶黄素 氧化应激 凋亡 核因子E2相关因子2/血红素加氧酶1/NADPH奎宁氧化还原酶1信号通路 实验研究 

分 类 号：TS201.4[轻工技术与工程—食品科学]