心脏Nmnat过表达联合耐力运动对果蝇高脂膳食诱发脂毒性心肌症的影响  

Effects of Cardiac Nmnat Overexpression Combined with Endurance Exercise on High-fat Diet Induced Lipotoxic Cardiomyopathy in Drosophila

作　　者：文登台 郑澜[1] 陆凯 刘宪楚[1] 后文其 WEN Dengtai;ZHENG Lan;LU Kai;LIU Xianchu;HOU Wenqi(Key Laboratory of Physical Fitness and Exercise Rehabilitation,Hunan Normal University Changsha,Hunan 410012,China;Department of Physical Education,Ludong University,Yantai,Shandong 264025,China)

机构地区：[1]湖南师范大学体适能与运动康复湖南省重点实验室,湖南长沙410012 [2]鲁东大学,山东烟台264025

出　　处：《首都体育学院学报》2020年第1期87-96,共10页Journal of Capital University of Physical Education and Sports

基　　金：国家自然科学基金资助项目（31671243）;国家青年基金资助项目（81600320）

摘　　要：利用UAS/hand-gal4系统构建果蝇心脏Nmnat基因特异性过表达,研究心脏Nmnat基因过表达联合耐力运动对高脂膳食诱发脂毒性心肌症的影响,并分析其分子机制。方法:雄性w^1118和NmnatUAS系果蝇分别与雌性hand-Gal4杂交,收集F1代心脏Nmnat正常表达(hand>w^1118)和过表达(hand>Nmnat)处女蝇,饲养至15 d后开始进行耐力运动(E)和高脂膳食(HFD),并分为hand>w^1118、hand>w^1118+E、hand>w^1118+HFD、hand>w^1118+HFD+E、hand>Nmnat、hand>Nmnat+E、hand>Nmnat+HFD、hand>Nmnat+HFD+E共8组,每组410只。ELISA检测心脏TAG、NAD+、SIR2蛋白去乙酰化、SOD活力和MDA水平,qRT-PCR检测心脏相关基因mRNA表达,M-mode检测心脏功能情况。结果:hand>w^1118+HFD+E果蝇心脏的Nmnat、dFoxo、bmm表达、NAD+水平、SIR2去乙酰化、SOD活力、缩短分数高于hand>w^1118+HFD(P<0.05或P<0.01),且d FAS和Col4al表达、TAG和MDA水平、心率和纤维性振颤低于hand>w^1118+HFD(P<0.05或P<0.01);hand>Nmnat果蝇心脏各指标与hand>Nmnat+HFD相比较未见显著差异(P>0.05);hand>Nmnat+HFD+E果蝇心脏的Nmnat、dFoxo、bmm表达、心脏NAD+水平、SIR2去乙酰化、SOD活力、心脏缩短分数高于hand>Nmnat+HFD(P<0.05或P<0.01),且dFAS和Col4al表达\TAG和MDA水平、心率和纤维性振颤低于hand>Nmnat+HFD(P<0.05或P<0.01)。结论:心脏Nmnat过表达和耐力运动均能抵抗果蝇高脂膳食诱发的心脏脂质过度堆积、氧化损伤、减弱心脏收缩能力和增加纤维性振颤,其分子机制与心脏Nmnat/NAD+/SIR2/dFoxo通路活性增强有关。心脏Nmnat基因过表达联合耐力运动能更好地降低脂毒性心肌症的发生概率,其机制与二者联合对心脏Nmnat/NAD+/SIR2/dFoxo通路活性上调的叠加作用有关。Objective:Constructing the specific overexpression of Nmnat gene in the heart of fruit flies using UAS/hand-gal4 system to study the effect of the overexpression of Nmnat gene in the heart combined with endurance exercise on lipid toxicity induced by high-fat diet,and to analyze the molecular mechanism.Methods:Male w^1118 and Nmnat-UAS fruit flies were respectively hybridized with female hand-gal4.Collecting the normal expression of heart Nmnat(hand>w^1118)and the overexpression(hand>Nmnat)of F1 generation female flies.After feeding for 15 days,they began to perform endurance exercise(E)and high-fat diet(HFD).They were divided into hand>w^1118,hand>w^1118+E,hand>w^1118+HFD,hand>w^1118+HFD+E,hand>Nmnat,hand>Nmnat+E,hand>Nmnat+HFD,hand>Nmnat+HFD+E,410 flies in each group.The levels of cardiac TAG,NAD+,SIR2 protein deacetylation,SOD activity and MDA were detected by ELISA.The expression of cardiac related genes by RT-PCR;M-mode detects cardiac function.Results:the expression of Nmnat,d Foxo,bmm,NAD+level,SIR2 deacetylation,SOD activity and shortened fraction of hand>w^1118+HFD+E group were significantly higher than that of hand>w^1118+HFD group(P<0.05 or P<0.01),but the expression of d FAS,Col4 al,TAG and MDA level,heart rate and fibrillation were significantly lower than that of hand>w^1118+HFD group(P<0.05 or P<0.01).There was no significant difference between the heart hand>Nmnat group and hand>Nmnat+HFD group(P>0.05).The expression of Nmnat,d Foxo,bmm,NAD+,SIR2 deacetylation,SOD activity and shortened heart fraction in the hand>Nmnat+HFD+E group were significantly higher than those in the hand>Nmnat+HFD group(P<0.05 or P<0.01),but the expression of d FAS,Col4 al,TAG and MDA levels,heart rate and fibrillation were significantly lower than those in the hand>Nmnat+HFD group(P<0.05 or P<0.01).Conclusion:both overexpression of cardiac Nmnat and endurance exercise can prevent excessive accumulation of cardiac lipids,oxidative damage,decreased cardiac contractility and increased fibrillation induced by high-fat diet.Th

关 键 词：耐力运动 高脂膳食 脂毒性心肌症 Nmnat 

分 类 号：G804.2[文化科学—运动人体科学]