双氢青蒿素通过抑制蛋白激酶B和NF-κκB P65磷酸化减轻缺血再灌注导致的小鼠心肌损伤  

Dihydroartemisinin alleviates myocardial damage of mice with myocardial ischemia/reperfusion injury by inhibiting phosphorylation of protein kinase B and NF-κκB P65

作　　者：崔勤涛[1] 王学惠[2] 刘永强[1] 刘晓晨[1] 段长恩 CUI Qin-tao;WANG Xue-hui;LIU Yong-qiang;LIU Xiao-chen;DUAN Chang-en(Department of Cardiovascular Surgery,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453100,China;Department of Cardiology,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453100,China)

机构地区：[1]新乡医学院第一附属医院心血管外科,河南新乡453100 [2]新乡医学院第一附属医院心血管内科,河南新乡453100

出　　处：《中国药理学与毒理学杂志》2021年第4期259-264,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：河南省2018年科技发展计划项目(1821023-10182)。

摘　　要：目的探讨双氢青蒿素(DHA)对心肌缺血再灌注(I/R)小鼠心肌氧化应激损伤作用及其机制。方法采用左冠状动脉前降支结扎法制备心肌I/R小鼠模型。结扎前20 min,模型+DHA 12.5,25.0和50.0 mg·kg^(-1)组小鼠分别ig给予相应剂量DHA,假手术组和I/R模型组ig给予等量生理盐水,每天1次,连续7 d。检测并记录小鼠心率(HR)、左心室射血分数(LVEF)和左室收缩末期容积(LVESV);用试剂盒测定心肌组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;ELISA检测小鼠血清中肌红蛋白(Mb)、心肌肌钙蛋白I(cTnI)和肌酸激酶同工酶(CK-MB)蛋白表达水平;HE染色观测心肌组织损伤;末端标记法(TUNEL)染色观测心肌细胞凋亡;Western印迹法检测心肌组织中Bax和Bcl-2蛋白表达水平及Akt和NF-κB P65磷酸化水平。结果心功能超声检测结果显示,与I/R模型组相比,I/R+DHA 25.0和50.0 mg·kg^(-1)组小鼠HR和LVEF明显升高(P<0.05,P<0.01),LVESV明显降低(P<0.05,P<0.01)。氧化应激检测结果显示,与I/R模型组比较,I/R+DHA 25.0和50.0 mg·kg^(-1)组小鼠心肌细胞中SOD活性显著升高(均P<0.05),MDA含量显著降低(均P<0.05)。ELISA检测结果显示,与I/R模型组比较,I/R+DHA 25.0和50.0 mg·kg^(-1)组小鼠血清中Mb,cTnI和CK-MB含量均显著降低(P<0.05,P<0.01)。HE染色结果显示,I/R模型组小鼠心肌组织结构紊乱,可见大量炎性浸润细胞;I/R+DHA 25.0和50.0 mg·kg^(-1)组小鼠心肌组织形态正常,未见炎性浸润细胞。TUNEL检测结果显示,I/R模型组小鼠心肌组织中可见大量凋亡细胞,I/R+DHA 25.0和50.0 mg·kg^(-1)组小鼠心肌组织中仅见极少凋亡细胞。Western印迹结果显示,与I/R模型组相比,I/R+DHA 25.0和50.0 mg·kg^(-1)组小鼠心肌组织中Bax/Bcl-2比值、Akt和NF-κB P65磷酸化水平均显著降低(P<0.05,P<0.01)。结论DHA可缓解心肌I/R小鼠氧化应激和心肌损伤,其机制可能与抑制Akt和NF-κB P65磷酸化有关。OBJECTIVE To investigate the effects and mechanisms of dihydroartemisinin(DHA)on oxidative stress injury in myocardial ischemia/reperfusion(I/R)mice.METHODS Myocardial I/R mouse model was established by ligating the left anterior descending coronary artery.20 min before that,the mice in the model+DHA 12.5,25.0 and 50.0 mg·kg^(-1) groups were ig given corresponding doses of DHA respectively,and the sham group and I/R model group were given the same amount of saline once a day for 7 d.Heart rate(HR),left ventricular ejection fraction(LVEF)and left ventricular end-systolic volume(LVESV)in each group were detected and recorded.The activity of superoxide dismutase(SOD)and contents of malondiadehyde(MDA)were determined by kits.The expression levels of myoglobin(Mb),cardiac troponin I(c Tn I)and creatine kinase isoenzyme(CK-MB)protein in serum of mice were measured by ELISA.Hematoxylin-eosin(HE)and end labeling(TUNEL)staining were used to observe the pathological damage and apoptosis of myocardial tissues.Western blotting was used to detect the protein expression levels of Bax and Bcl-2 as well as the phosphorylation levels of Akt and NF-κB P65 in myocardial tissues.RESULTS The results of cardiac function ultrasound examination showed that compared with the I/R model group,HR and LVEF of mice in the I/R+DHA 25.0 and 50.0 mg·kg^(-1) groups were significantly increased(P<0.05,P<0.01),while the LVESV was significantly reduced(P<0.05,P<0.01).The oxidative stress test results showed that compared with the I/R model group,the SOD content in the myocardial cells of mice in the I/R+DHA 25.0 and 50.0 mg·kg^(-1) groups was significantly increased(P<0.05),while the MDA content was significantly decreased(P<0.05).ELISA test results showed that compared with the I/R model group,the serum levels of Mb,cTnI and CK-MB in the I/R+DHA 25.0 and50.0 mg·kg^(-1) groups were significantly reduced(P<0.05,P<0.01).The results of HE staining showed that the myocardial tissue structure of mice in the I/R model group was disorderly,and a large

关 键 词：双氢青蒿素 缺血再灌注 心肌损伤 氧化应激 

分 类 号：R966[医药卫生—药理学]