吗啡预处理对缺氧/复氧诱发的心肌细胞线粒体动力学变化的影响  

Effects of morphine pretreatment on mitochondrial dynamics change induced by hypoxia/reoxygenation in myocardial cells

作　　者：高亚云 王蕴琦 张熙 贡美娜 逄晓倩 田炜 徐菁蔓 GAO Ya-yun;WANG Yun-qi;ZHANG Xi;GONG Mei-na;PANG Xiao-qian;TIAN Wei;XU Jing-man(School of Public Health,North China University of Science and Technology,Tangshan 063210,Hebei Pirovince,China;Laboratory Animal Center,North China University of Science and Technology,Tangshan 063210,Hebei Pirovince,China;Hebei Key Laboratory of Organ Fibrosis,Tangshan 063210,Hebei Pirovince,China)

机构地区：[1]华北理工大学,公共卫生学院,河北唐山063210 [2]华北理工大学,实验动物中心,河北唐山063210 [3]河北省器官纤维化重点实验室,河北唐山063210

出　　处：《中国临床药理学杂志》2024年第14期2023-2027,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨吗啡(Mor)对缺氧/复氧(H/R)诱发的心肌H9c2细胞线粒体动力学变化的影响。方法将心肌H9c2细胞分为空白组(正常培养)、模型组(H/R处理)、对照组(5μmol·L^(-1) Mor处理)、实验组(H/R+5μmol·L^(-1) Mor处理)。用透射电子显微镜(TEM)观察线粒体的形态,用活性氧(ROS)、四甲基罗丹明乙酯(TMRE)、线粒体复合体Ⅰ和Ⅲ活性检测试剂盒分别检测ROS含量、线粒体膜电位(MMP)、复合体Ⅰ和Ⅲ活性,用蛋白质印迹法检测线粒体动力学相关蛋白GTP酶动力蛋白相关蛋白1(Drp1)、细胞色素氧化酶Ⅳ亚型(COXⅣ)和线粒体外膜转位酶20(TOM20)蛋白的表达水平。结果实验组细胞核膜光滑完整,线粒体大小一致,嵴排列整齐,空泡化减少,线粒体基质电子密度增加,自噬体数量减少。空白组、模型组、对照组和实验组细胞的ROS含量分别为1.03±0.04、1.53±0.10、1.06±0.06和1.10±0.11,MMP分别为1.00±0.15、0.80±0.16、1.06±0.19和1.00±0.19,复合体Ⅰ活性分别为1.00±0.08、2.28±0.82、1.05±0.26和1.13±0.37,复合体Ⅲ活性分别为1.00±0.09、2.13±0.38、0.83±0.22和0.96±0.11,Drp1蛋白相对表达水平分别为1.00±0.14、1.27±0.07、0.97±0.21和0.93±0.17,线粒体裂变蛋白1(Fis1)蛋白相对表达水平分别为1.00±0.16、1.33±0.18、1.17±0.25和0.99±0.05,COXⅣ蛋白相对表达水平分别为1.00±0.25、0.62±0.08、0.79±0.26和0.97±0.16,TOM20蛋白相对表达水平分别为1.00±0.13、0.67±0.15、0.75±0.13和0.89±0.05。模型组的上述指标与空白组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01,P<0.001,P<0.0001);实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01,P<0.001,P<0.0001)。结论吗啡可能通过抑制心肌线粒体自噬和分裂,以及降低呼吸链复合体Ⅰ和Ⅲ的活性减轻氧化应激损伤,进而维持线粒体动力学稳态,减轻H/R诱发的心肌细胞损伤。Objective The aim of the study was to investigate how morphine(Mor)effects mitochondrial dynamics change of H9c2 induced by hypoxia/reoxygenation(H/R).Methods Myocardial H9c2 cells were divided into blank group(without treatment),model group(H/R treatment),control group(5μmol·L^(-1)Mor treatment)and experimental group(H/R+5μmol·L^(-1)Mor treatment).The content of reactive oxygen species(ROS),mitochondrial membrane potential(MMP),and complex ofⅠandⅢactivity were detected using ROS tetramethylrhodamine ethyl ester(TMRE),and mitochondrial complex ofⅠandⅢactivity detection kits,respectively.The morphology of mitochondria and lysosomes was observed by transmission electron microscope electron microscopy(TEM);Western blot was used to detect the expression of GTPase kinetic protein 1(Drp1),cytochrome c oxidaseⅣ(COXⅣ)and transporters of the outer mitochondrial membrane(TOM20).Results The nuclear membrane was smooth and complete;the mitochondrial size was consistent;the crest arrangement was neat;vacuolization was reduced or even disappeared;the mitochondrial matrix electron density was increased;the number of autophagosomes was decreased in the experimental group.The contents of ROS in blank group,model group,control group and experimental group were 1.03±0.04,1.53±0.10,1.06±0.06 and 1.10±0.11;MMP were1.00±0.15,0.80±0.16,1.06±0.19 and 1.00±0.19;the activities of complex ofⅠwere 1.00±0.08,2.28±0.82,1.05±0.26 and 1.13±0.37;the activities of complex ofⅢwere 1.00±0.09,2.13±0.38,0.83±0.22 and 0.96±0.11;the expression of Drp1 protein were 1.00±0.14,1.27±0.07,0.97±0.21 and0.93±0.17;the expression of fission protein 1(Fis1)protein were 1.00±0.16,1.33±0.18,1.17±0.25 and0.99±0.05;the expression of COXⅣprotein were 1.00±0.25,0.62±0.08,0.79±0.26 and 0.97±0.16;the expression of TOM20 protein were 1.00±0.13,0.67±0.15,0.75±0.13 and 0.89±0.05.The above indexes of model group were significantly different from those of blank group(P<0.05,P<0.01,P<0.001,P<0.0001).The above indexes of ex

关 键 词：吗啡 心肌缺氧/复氧损伤 线粒体动力学 活性氧 

分 类 号：R971.2[医药卫生—药品]