二氟甲基鸟氨酸对心肌肥厚模型线粒体动力学及氧化应激的影响  

Effects of α-difluoromethylornithine on mitochondrial dynamics and oxidative stress in the model of cardiac hypertrophy

作　　者：赵宇 王璐璐 任珊 林岩[1] 张宁宁 王玉春[1] 贾伟伟 孙晓娇 ZHAO Yu;WANG Lu-lu;REN Shan;LIN Yan;ZHANG Ning-ning;WANG Yu-chun;JIA Wei-wei;SUN Xiao-jiao(Basic Medical College,Qiqihar Medical University,Qiqihar 161006,Heilongjiang Province,China)

机构地区：[1]齐齐哈尔医学院基础医学院,黑龙江齐齐哈尔161006

出　　处：《中国临床药理学杂志》2022年第14期1659-1663,共5页The Chinese Journal of Clinical Pharmacology

基　　金：黑龙江省省属本科高校基本科研业务费基金资助项目(2019-KYYWF-1226)。

摘　　要：目的探讨二氟甲基鸟氨酸(DFMO)对异丙肾上腺素(ISO)诱导的心肌肥厚模型线粒体动力学改变及对氧化应激的影响。方法将Wistar大鼠随机分为空白组、模型组和实验组,每组10只。空白组给予等量0.9%NaCl连续皮下注射1周;模型组给予5 mg·kg^(-1) ISO连续皮下注射1周复制心肌肥厚模型;实验组给予2%DFMO水溶液,连续给药10 d。比较3组大鼠的心脏指数和心肌组织谷胱甘肽过氧化物酶(GSH-Px)的活性。将H9C2心肌细胞分为空白组、模型组、实验组和对照组。空白组给予磷酸盐缓冲溶液处置;模型组给予10μmol·L^(-1) ISO作用48 h;实验组在加入10μmol·L^(-1) ISO前24 h加入1 mmol·L^(-1) DFMO处置;对照组在加入10μmol·L^(-1) ISO前24 h加入0.5 mmol·L^(-1)腐胺。用蛋白质印迹(Western blot)法检测线粒体分裂蛋白(DRP1)和线粒体融合蛋白(MFN2)蛋白的表达水平,用单管多功能检测仪测定细胞内钙离子浓度,用试剂盒测定心肌细胞线粒体超氧化物的水平。结果动物实验:空白组、模型组和实验组的心脏指数分别为(3.58±0.59),(3.74±1.49)和(3.62±0.76)mg·g^(-1),GSH-Px含量分别为(183.12±29.28),(149.60±24.24)和(173.29±23.59)U·mg^(-1),实验组的上述指标与模型组比较,差异均有统计学意义(均P<0.05)。细胞实验:空白组、模型组、实验组和对照组心肌细胞的DRP1蛋白相对表达水平分别为1,2.26±0.31,1.09±0.19和2.28±0.42,MFN2蛋白相对表达水平分别为1,0.77±0.13,1.16±0.02和0.95±0.11,细胞内钙离子浓度(荧光强度)分别为(66.10±4.54),(80.90±10.18),(61.60±5.30)和(73.00±8.06)abs·unit,实验组的上述指标与模型组比较,差异均有统计学意义(P<0.05或P<0.01)。结论DFMO对心肌肥厚有保护作用,其机制与抑制线粒体分裂和氧化应激有关。Objective To investigate the effects ofα-difluoromethylornithine(DFMO)on changes of mitochondrial dynamics and oxidative stress in cardiac hypertrophy model induced by isoproterenol(ISO).Methods Wistar rats were randomly divided into 3 groups:blank group,model group and experimental group,10 rats in each group.The rats in blank group were treated with 0.9%NaCl for 1 week;model rats were established by subcuta-neously 5 mg·kg^(-1) ISO,once a day for 1 week;the experimental group was given 2%DFMO dissolved in water for 10 days.The heart index and myocardial tissue glutathione peroxidase(GSH-Px)activity of the 4 groups of rats were compared.H9C2 cardiomyocytes were divided into blank,model,experimental and control groups.The blank group was treated with phosphate buffered saline;the model group was treated with 10μmol·L^(-1) ISO for 48 hours;the experimental group was treated with 1 mmol·L^(-1) DFMO 24 hours before the addition of 10μmol·L^(-1) ISO;the control group was treated with 0.5 mmol·L^(-1)putrescine 24 h before the addition of 10μmol·L^(-1) ISO.The protein expression levels of mitochondrial fission protein(DRP1)and mitochondrial fusion protein(MFN2)were measured by Western blot.Calcium concentration was measured by single-tube multifunctional detector.The level of mitochondrial superoxide in cardiomyocytes was detected by kit.Results Animal experiment:The heart indexes in blank,model and experimental groups were(3.58±0.59),(3.74±1.49)and(3.62±0.76)mg·g^(-1),the levels of GSH-Px were(183.12±29.28),(149.60±24.24)and(173.29±23.59)U·mg^(-1).The differences between the experimental group and the model group were statistically significant(all P<0.05).Cell experiment:The DRP1 protein expression level in the blank,model,experimental and control groups were 1,2.26±0.31,1.09±0.19 and 2.28±0.42,the MFN2 protein expression level were 1,0.77±0.13,1.16±0.02 and 0.95±0.11,the calcium concentrations in the four groups of cardiomyocytes were(66.10±4.54),(80.90±10.1),(61.60±5.30)and(73.00±8.06)abs

关 键 词：二氟甲基鸟氨酸 心肌肥厚 线粒体动力学 氧化应激 

分 类 号：R97[医药卫生—药品]