Omega-6型多不饱和脂肪酸增加急性心肌缺血损伤“易感性”的机制研究  

Mechanism of omege-6 PUFA increasing the susceptibility to acute myocardial ischemia injury

作　　者：刘江涵子 马晓慧 袁天慧[5] 孙万阳[1,2,3] 李怡芳[1,2,3] 栗原博[1,2,3] 何蓉蓉[1,2,3] LIU Jiang-han-zi;MA Xiao-hui;YUAN Tian-hui;SUN Wan-yang;LI Yi-fang;HIROSHI Kurihara;HE Rong-rong(Guangdong Engineering Research Center of Chinese Medicine and Disease Susceptibility,Jinan University,Guangzhou 510632,China;Institute of Traditional Chinese Medicine and Natural Products,College of Pharmacy,Jinan University,Guangzhou 510632,China;Guangdong Provincial Key Laboratory of Pharmacodynamic Substances of Traditional Chinese Medicine and Innovative Drugs,Jinan University,Guangzhou 510632,China;Institute of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830054,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]暨南大学,广东省疾病易感性及中医药研发工程技术研究中心,广东广州510632 [2]暨南大学药学院,中药及天然药物研究所,广东广州510632 [3]暨南大学,广东省中药药效物质基础及创新药物研究重点实验室,广东广州510632 [4]新疆医科大学中医学院,新疆乌鲁木齐830054 [5]广州中医药大学第一附属医院,广东广州510405

出　　处：《药学学报》2022年第6期1657-1663,共7页Acta Pharmaceutica Sinica

基　　金：国家重点研发计划项目(2017YFC1700404);国家自然科学基金资助项目(82125038,81904068);珠江学者计划项目(GDUPS2019);珠江创新团队项目(2017BT01Y036)。

摘　　要：脂质代谢紊乱是冠心病的常见病理特征。虽然增加不饱和脂肪酸(PUFA)的摄入一直以来被认为可以有效预防冠心病,但是越来越多的临床试验却否定了该观点。PUFA在冠心病中的具体作用机制尚不明确。本实验通过建立omega-6型PUFA负荷急性心肌缺血的动物模型,来探讨PUFA对心肌缺血损伤的影响及其潜在机制。实验方案及程序均符合动物使用和护理的伦理原则,并已获暨南大学动物实验伦理委员会批准。实验采用亚油酸(LA)连续灌胃14天后,大剂量腹腔注射异丙肾上腺素(ISO)诱导急性心肌缺血以建立动物模型。24 h后,采用小动物超声影像系统检测心脏功能。随后,采集血清及心肌组织样本进行心肌酶、磷脂组学分析、试剂盒含量测定和Western blot检测。心脏功能结果表明,与单纯LA组比较,LA负荷ISO小鼠的左心室射血分数(EF%)和左心室缩短分数(FS%)显著降低,天门冬氨酸转移酶(AST)、肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)显著增加。磷脂组学分析表明,与ISO组比较,LA负荷ISO小鼠心肌组织中的PUFA显著增加,且氧化型磷脂酰乙醇胺(ox-PE)的含量变化最为显著。还原型辅酶Ⅱ(NADPH)、谷胱甘肽(GSH)、丙二醛(MDA)试剂盒测定结果表明,与单纯LA组比较,LA负荷ISO小鼠心肌组织中的GSH和NADPH的含量显著降低,ox-PE的产物MDA显著增加。Western blot结果显示,花生四烯酸12/15-脂氧合酶(ALOX15)蛋白表达随缺血造模显著增加,亚油酸负荷后4-羟基壬烯醛(4-HNE)的表达显著增加。本研究结果表明,ALOX15介导的ox-PE产生可能是LA摄入过量加剧心肌缺血损伤“易感性”的关键机制。本实验为PUFA是否可作为一种有效预防冠心病的代替疗法策略提供实验依据,更为冠心病的早期预防策略提供新的干预靶标。The abnormal lipids metabolism is a critical pathological feature of coronary heart disease(CHD).Additional supplemental intake of polyunsaturated fatty acid(PUFA)has long been considered to be an effective strategy for preventing CHD,but more and more clinical trials have denied this view.Still,it is ambiguity for the specific mechanism of PUFA in CHD.The experimental programs are compliant with ethical principles for animal use and have been approved by the Animal Experiment Ethics Committee of Jinan University.In the present study,we established an animal model by intake of omega-6 PUFA combined acute myocardial ischemia to explore the mechanism of CHD.Intragastric administration of linoleic acid(LA)for 14 days,intraperitoneal injection of isoprenaline(ISO)was applied to induce acute myocardial ischemia for the animal model establishment.The animal ultrasound imaging system was used to detect cardiac function in vivo after ISO injection for 24 h.Serum and heart tissue samples were collected for the myocardial enzyme,phospholipidomics analysis and molecular biological detection.Compared to the LA group,the cardiac function showed that the left ventricular ejection fraction(EF%)and the left ventricular shortening fraction(FS%)decreased,aspaetate aminotransferase(AST),creatine kinase isoenzyme(CK-MB),and lactate dehydrogenase(LDH)increased in the LA+ISO mice.Compared to the ISO group,the phospholipidomics analysis showed that the PUFAs significantly were raised in the LA+ISO myocardium,and the content of oxidized phosphatidylethanolamine(ox-PE)changed most remarkable.Compared with the ISO group,the molecular biology detection showed that glutathione(GSH)and nicotinamide adenine dinucleotide phosphate(NADPH)were depleted,the end-products of ox-PE were increased,and the level of arachidonic acid 12/15-lipoxygenase(ALOX15)protein expression increased obviously.We suggest that ALOX15 mediated phospholipid peroxidation might be the critical mechanism of LA increased the susceptibility of myocardial ischemia injury.Thi

关 键 词：不饱和脂肪酸 心肌缺血 易感性 花生四烯酸12/15-脂氧合酶 脂质过氧化 

分 类 号：R966[医药卫生—药理学]