热休克蛋白A12B介导COX-2表达调控心肌缺血再灌注损伤中心肌细胞线粒体自噬的作用研究  

Study on the role of heat shock protein A12B mediated COX-2 expression in regulating mitochondrial autophagy of myocardial cells in myocardial ischemia-reperfusion injury

作　　者：王书君 谭慧琼 袁杰[3] WANG Shu-jun;TAN Hui-qiong;YUAN Jie(Department of Geriatric Medicine,First Affiliated Hospital of Hainan Medical College,Haikou Hainan 570102,China;Department of Cardiovascular Medicine,Fuwai Hospital,Chinese Academy of Medical Sciences,Beijing 100037,China;Department of Cardiology,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi Xinjiang 830001,China)

机构地区：[1]海南医学院第一附属医院老年医学科,海南海口570102 [2]中国医学科学院阜外医院心血管内科,北京100037 [3]新疆维吾尔自治区人民医院心内科,新疆乌鲁木齐830001

出　　处：《局解手术学杂志》2022年第9期759-765,共7页Journal of Regional Anatomy and Operative Surgery

基　　金：海南省卫生健康行业科研项目(20A200405)。

摘　　要：目的 探讨热休克蛋白A12B(HSPA12B)对心肌缺血再灌注(MI/R)损伤中心肌细胞线粒体自噬的影响及可能的分子机制。方法 将40只SD大鼠随机分为假手术组、模型组、p LVX-NC组和p LVX-HSPA12B组,每组10只。其中,假手术组大鼠仅暴露心脏,不进行MI/R处理;模型组大鼠给予心肌内注射0.9%氯化钠注射液20μL,7 d后构建MI/R模型;p LVX-NC组大鼠给予心肌内注射20μL p LVX-NC慢病毒液,7 d后构建MI/R模型;p LVX-HSPA12B组大鼠给予心肌内注射20μL p LVX-HSPA12B慢病毒液,7 d后构建MI/R模型。ELISA检测各组大鼠血清肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)含量,HE染色观察各组大鼠心肌组织病理学形态,TUNEL染色检测各组大鼠心肌组织内细胞凋亡情况,免疫组织化学染色检测各组大鼠心肌组织内环氧化酶-2(COX-2)和微管相关蛋白1轻链3β(LC3β)表达,Western blot检测各组大鼠心肌组织内LC3Ⅱ/LC3Ⅰ比值及Parkin、PINK1蛋白的表达水平。结果 与假手术组比较,模型组大鼠血清中CK-MB和LDH的含量均升高,心肌组织损伤较为严重,心肌组织内TUNEL阳性细胞率升高,COX-2阳性表达率上升,而LC3β阳性表达率下降,LC3Ⅱ/LC3Ⅰ比值下降,Parkin、PINK1蛋白的表达水平均下调(P <0.05);与模型组和p LVX-NC组比较,p LVX-HSPA12B组大鼠血清中CK-MB和LDH的含量均降低,心肌组织损伤现象明显减轻,心肌组织内TUNEL阳性细胞率降低,COX-2阳性表达率下降,同时LC3β阳性表达率上升,LC3Ⅱ/LC3Ⅰ比值升高,Parkin、PINK1蛋白的表达水平均上调(P <0.05)。结论 HSPA12B能够通过增加心肌细胞线粒体自噬途径来缓解MI/R模型大鼠心肌损伤,其作用机制可能与抑制COX-2表达有关。Objective To investigate the effect of heat shock protein A12B( HSPA12B) on mitochondrial autophagy of myocardial cells in myocardial ischemia-reperfusion( MI/R) injury and its possible molecular mechanism. Methods A total of 40 SD rats were randomly divided into the sham operation group,the model group,the p LVX-NC group and the p LVX-HSPA12B group,with 10 rats in each group.Among them,rats in the sham operation group only exposed the heart without MI/R treatment,rats in the model group were given intramyocardial injection of 20 μL of 0. 9% sodium chloride injection and then the MI/R model was constructed after 7 days;rats in the p LVX-NC group were given intramyocardial injection of 20 μL p LVX-NC lentivirus and then the MI/R model was constructed after 7 days;rats in the p LVXHSPA12B group were given intramyocardial injection of 20 μL p LVX-HSPA12B lentivirus and then the MI/R model was constructed after7 days. ELISA was used to determine the levels of serum creatine kinase isoenzymes( CK-MB) and lactate dehydrogenase( LDH) of rats in each group,HE staining was used to observe the pathological morphology of myocardial tissues of rats in each group,TUNEL staining was used to detect the cell apoptosis in myocardial tissues of rats in each group,the immunohistochemical staining was used to detect the expressions of cyclooxygenase-2( COX-2) and microtubule-associated protein 1 light chain 3 beta( LC3β) in myocardial tissues of rats in each group,Western blot was used to determine the ratio of LC3Ⅱ/LC3Ⅰ and the expression levels of Parkin and PINK1 proteins in myocardial tissues of rats in each group. Results Compared with the sham operation group,the levels of serum CK-MB and LDH in the model group increased,the damage of myocardial tissues was more severe,the TUNEL positive cell rate in the myocardial tissues increased,and the COX-2 positive expression rate increased,while the LC3β positive expression rate decreased,the ratio of LC3 Ⅱ/LC3 Ⅰ decreased,and the expression levels of Parkin and PINK1 prot

关 键 词：心肌缺血再灌注 热休克蛋白A12B 环氧化酶-2 线粒体 自噬 

分 类 号：R542.2[医药卫生—心血管疾病]