摘要
目的 研究干扰肥胖小鼠心肌CPT1b的表达对心肌活性氧簇（ROS）的影响.方法 4周龄的雄性C57小鼠,随机分为三组,分别为正常对照组（N-mock）、肥胖对照组（O-mock）及肥胖干预组（O-CPT1b）,对肥胖组小鼠进行高脂饮食造肥胖模型.6周龄时,向干预组小鼠心肌注射靶向CPT1b（O-CPT1b）或靶向无关基因（N-mock、O-mock）的重组慢病毒.继续喂养10周后,取小鼠心肌,Western blot检测CPT1b的蛋白表达量;行油红O染色检测心肌组织中性脂质含量;使用冷冻切片染色法及流式细胞术检测心肌细胞中ROS.结果 RT-PCR和Western blot结果显示,携带CPT1b基因的短发夹RNA显著下调了O-CPT1b小鼠心肌组织中CPT1b的表达;肥胖引起心肌组织内中性脂质蓄积增多,下调CPT1b的表达增加了心肌内中性脂质的含量;高脂饮食还导致心肌ROS的含量显著增加,而下调CPT1b的表达可以改善甚至逆转这一进程.结论 CPT1b在肥胖小鼠心肌的ROS生成增多中起重要作用,下调小鼠心肌组织的CPT1b表达或许会通过减轻心肌氧化应激而改善肥胖性心脏重构.
Objective To study the effect of interfering with the expression of CPT1b in myocardial tis- sues of obese mice on reactive oxygen species metabolism. Methods Four -week-old male C57 mice were ran- domly divided into three groups： normal control group （N-mock）, obese control group （O-mock） and obesity in- tervention group （O-CPT1b）. Mice were subjected to a high-fat diet to induce obesity models. At 6 weeks of age, mice in the intervention group were injected with a recombinant lentivirus targeting CPT1b （O-CPT1b） or an un- targeted gene （N-mock, O-mock）. After 10 weeks of continuous feeding, the left ventricular tissues of the mice was obtained, the mRNA and protein expression of CPT1b was detected by RT-PCR and Western blot, the fat con- tent in myocardium was detected by oil red O staining, and the myocardial cells were detected by frozen section staining and flow cytometry. Results RT-PCR and Western blot showed that the short hairpin RNA（shRNA） car- rying CPT1b gene significantly down-regulated the expression of CPT1b in myocardial tissue of O-CPT1b mice; Obesity increased the accumulation of fat in myocardial tissue, down-regulating the expression of CPT1b can in- crease myocardial fat content; high-fat diet also led to a significant increase in myocardial ROS content, and down-regulation of CPT1b expression can ameliorate or even reverse this process. Conclusion CPT1b plays an important role in the increase of ROS production caused by high fatty acid metabolism. Down-regulation ofCPT1b expression in myocardium may ameliorate myocardial oxidative stress.
作者
罗强
张逸杰
包明威
关红菁
LUO Qiang;ZHANG Yi-jie;BAO Ming-wei(Department of Cardiology,Renmin Hospital of Wuhan Uni-versity,Cardiovascular Research Institute of Wuhan University,Hubei Key Laboratory of Cardiology,Wuhan 430060,China)
出处
《中国心血管病研究》
CAS
2018年第8期761-766,共6页
Chinese Journal of Cardiovascular Research
基金
国家自然科学基金青年科学基金资助项目(项目编号:81500668)
国家自然科学基金资助项目(面上项目)(项目编号:81570460)
湖北省卫生计生科研基金(项目编号:WJ2017Z003)
