过表达BMAL1对脓毒症小鼠心肌损伤的保护作用及机制研究-东南大学学报医学版

过表达BMAL1对脓毒症小鼠心肌损伤的保护作用及机制研究

单位：新疆维吾尔自治区人民医院重症医学科, 新疆乌鲁木齐 830001

关键词：脓毒症心肌损伤心肌细胞大脑和肌肉芳香烃受体核转运蛋白样蛋白1 线粒体自噬小鼠

分类号：R-332; R631

出版年·卷·期（页码）：2024·43·第一期（40-46）

摘要：

目的: 探究生物钟基因大脑和肌肉芳香烃受体核转运蛋白样蛋白1(BMAL1)过表达对脓毒症小鼠心肌损伤中线粒体自噬途径的影响。方法: 将40只小鼠随机分为模型对照组、模型组、阴性对照(NC)组、BMAL1组,每组10只。超声心动图检测小鼠心功能指标左室短轴缩短率(LVFS)、左室射血分数(LVEF)、左室舒张末期内径(LVIDd)、左室收缩末期内径(LVIDs),ELISA检测血清肌酸激酶同工酶(CK-MB)、肌钙蛋白I(cTnI)水平,HE染色观察心肌组织病理变化,TUNEL染色检测心肌细胞凋亡,透射电镜观察心肌线粒体超微结构变化,Western blotting测定心肌组织中微管相关蛋白1轻链3(LC3)、BH3域自噬蛋白(Beclin1)、PTEN诱导激酶(Pink1)、E3泛素-连接酶活性帕金森病蛋白2(Parkin)、电压依赖性阴离子通道蛋白1(VDAC1)表达水平。结果: 与模型组和NC组比较,BMAL1组小鼠LVFS、LVEF升高(P<0.05),LVIDd、LVIDs减小(P<0.05),血清中CK-MB、cTnI降低(P<0.05),心肌组织病变得到改善,TUNEL阳性细胞率减少(P<0.05),线粒体损伤减轻,心肌组织中LC3Ⅱ/LC3Ⅰ值升高(P<0.05),Beclin1、Pink1、Parkin、VDAC1蛋白表达上调(P<0.05)。结论: BMAL1过表达能够明显改善脓毒症小鼠心功能,减轻心肌损伤,该作用可能与其促进线粒体自噬有关。

Objective: To explore the effect of circadian clock gene aromatic hydrocarbon receptor nuclear transport-like protein 1(BMAL1) overexpression on mitochondrial autophagy pathway in myocardial injury of sepsis mice. Methods: Forty mice were randomly divided into model control group,model group,negative control(NC) group and BMAL1 group,with 10 mice in each group. Left ventricular short-axis shortening rate(LVFS),left ventricular ejection fraction(LVEF),left ventricular end-diastolic diameter(LVIDd),left ventricular end-systolic diameter(LVIDs) were detected by echocardiography in mice,serum creatine kinase isoenzyme(CK-MB) and troponin I(cTnI) were detected by ELISA,pathological changes of myocardial tissue were observed by HE staining,myocardial apoptosis was detected by TUNEL staining,the ultrastructural changes of myocardial mitochondria were observed by transmission electron microscopy,the expression levels of microtubule-associated protein 1 light chain 3(LC3),BH3-domain autophagy protein(Beclin1),PTEN-induced kinase(Pink1),E3 ubiquitin-ligase active Parkinson's disease protein 2(Parkin) and voltage-dependent anion channel protein 1(VDAC1) in myocardial tissue were determined by Western blotting. Results: Compared with model group and NC group,LVFS and LVEF in BMAL1 group were increased(P<0.05),LVIDd and LVIDs were decreased(P<0.05),serum CK-MB and cTnI were decreased(P<0.05),myocardial tissue lesions were improved,the rate of TUNEL positive cells was decreased(P<0.05),the damage to mitochondria was reduced,LC3Ⅱ/LC3Ⅰ ratio in myocardial tissue was further increased(P<0.05),and Beclin1,Pink1,Parkin and VDAC1 protein expression levels were further up-regulated(P<0.05).Conclusion: Overexpression of BMAL1 can significantly improve cardiac function and alleviate myocardial injury in sepsis mice,which may be related to the promotion of mitochondrial autophagy.

参考文献：

[1] ZHOU B,ZHANG J,CHEN Y,et al.Puerarin protects against sepsis-induced myocardial injury through AMPK-mediated ferroptosis signaling[J].Aging(Albany NY),2022,14(8):3617-3632.
[2] WENG L,XU Y,YIN P,et al.China critical care clinical trials group(CCCCTG).National incidence and mortality of hospitalized sepsis in China[J].Crit Care,2023,27(1):84.
[3] BI C F,LIU J,YANG L S,et al.Research progress on the mechanism of sepsis induced myocardial injury[J].J Inflamm Res,2022,15:4275-4290.
[4] KHALID N,PATEL P D,ALGHAREEB R,et al.The effect of sepsis on myocardial function:a review of pathophysiology,diagnostic criteria,and treatment[J].Cureus,2022,14(6):e26178.
[5] PUELACHER C,GUALANDRO D M,GLARNER N,et al.Long-term outcomes of perioperative myocardial infarction/injury after non-cardiac surgery[J].Eur Heart J,2023,27:ehac798.
[6] RAY S,VALEKUNJA U K,STANGHERLIN A,et al.Circadian rhythms in the absence of the clock gene Bmal1[J].Science,2020,367(6479):800-806.
[7] LIN H,JI F,LIN K Q,et al.LPS-aggravated ferroptosis via disrupting circadian rhythm by Bmal1/AKT/p53 in sepsis-induced myocardial injury[J].Inflammation,2023,13:37046145.
[8] SJAASTAD F V,JENSEN I J,BERTON R R,et al.Inducing experimental polymicrobial sepsis by cecal ligation and puncture[J].Curr Protoc Immunol,2020,131(1):e110.
[9] 孙蒙奇,姚续,王辉,等.脓毒性心肌病增加脓毒症患者的全因院内病死率[J].中日友好医院学报,2023,37(2):81-84,102.
[10] FRENCKEN J F,VAN SMEDEN M,VAN DE GROEP K,et al.Etiology of myocardial injury in critically ill patients with sepsis:a cohort study[J].Ann Am Thorac Soc,2022,19(5):773-780.
[11] LIANG Q,XU H,LIU M,et al.Postnatal deletion of Bmal1 in cardiomyocyte promotes pressure overload induced cardiac remodeling in mice[J].J Am Heart Assoc,2022,11(13):e025021.
[12] DAKUP P P,PORTER K I,GAJULA R P,et al.The circadian clock protects against ionizing radiation-induced cardiotoxicity[J].FASEB J,2020,34(2):3347-3358.
[13] QIU Z,MING H,LEI S,et al.Roles of HDAC3-orchestrated circadian clock gene oscillations in diabetic rats following myocardial ischaemia/reperfusion injury[J].Cell Death Dis,2021,12(1):43.
[14] DEMINE S,RENARD P,ARNOULD T.Mitochondrial uncoupling:a key controller of biological processes in physiology and diseases[J].Cells,2019,8(8):795.
[15] LI A,GAO M,LIU B,et al.Mitochondrial autophagy:molecular mechanisms and implications for cardiovascular disease[J].Cell Death Dis,2022,13(5):444.
[16] POZNYAK A V,NIKIFOROV N G,WU W K,et al.Autophagy and mitophagy as essential components of atherosclerosis[J].Cells,2021,10(2):443.
[17] 刘润民,汪雅乔,吴克寒,等.NLRP3炎症体的活化与心肌缺血再灌注损伤和线粒体自噬[J].现代医学,2023,51(7):1006-1012.
[18] 李开,饶莉.线粒体自噬的分子生物学过程及其在心脏疾病中的作用[J].心血管病学进展,2022,43(3):222-224,228.
[19] PEÑA-MARTINEZ C,RICKMAN A D,HECKMANN B L.Beyond autophagy:LC3-associated phagocytosis and endocytosis[J].Sci Adv,2022,8(43):eabn1702.
[20] TRAN S,FAIRLIE W D,LEE E F.BECLIN1:protein structure,function and regulation[J].Cells,2021,10(6):1522.
[21] EIYAMA A,OKAMOTO K.PINK1/Parkin-mediated mitophagy in mammalian cells[J].Curr Opin Cell Biol,2015,33:95-101.
[22] WU Y,JIANG T,HUA J,et al.PINK1/Parkin-mediated mitophagy in cardiovascular disease:from pathogenesis to novel therapy[J].Int J Cardiol,2022,361:61-69.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录