玉米肽Leu-Pro-Phe对酒精诱导C2C12细胞能量代谢障碍与氧化应激的影响  

Effects of Corn Peptide Leu-Pro-Phe on Ethanol-Induced Mitochondrial Energy Metabolism Impairment and Oxidative Stress in C2C12 Cells

作　　者：余宗盛 黎新江 陈小娟 栗原博 毛新亮 YU Zongsheng;LI Xinjiang;CHEN Xiaojuan;KURIHARA Hiroshi;MAO Xinliang(Pefect(GuangDong)Co.,Ltd.,Zhongshan,Guangdong 528400,China;Perfect Life Sciences Research Institute Co.,Ltd.,Zhongshan,Guangdong 528400,China)

机构地区：[1]完美(广东)日用品有限公司,广东中山528400 [2]广东完美生命健康科技研究院有限公司,广东中山528400

出　　处：《今日药学》2023年第10期733-739,共7页Pharmacy Today

基　　金：广东省重点领域研发计划项目(2022B0202030001)。

摘　　要：目的探究玉米肽Leu-Pro-Phe(LPF)对酒精诱导C2C12细胞线粒体能量代谢障碍与氧化应激的影响。方法将C2C12细胞诱导分化为肌管后,给予乙醇(300 mmol·L^(-1))作用6 h构建酒精应激模型;同时给予LPF(10、20、40μmol·L^(-1))评价其对细胞的保护作用。采用ATP试剂盒与呼吸链复合物试剂盒检测C2C12细胞内的ATP的水平以及5种呼吸链复合酶的活性;采用罗丹明123染色检测线粒体的膜电位(mitochondrial membrane potential,MMP)。采用H2DCFDA探针检测细胞内活性氧(reactive oxygen species,ROS)的水平;采用试剂盒检测细胞内丙二醛(malondialdehyde,MDA)与谷胱甘肽(glutathione,GSH)的含量以及超氧化物歧化酶(superoxide dismutase,SOD)与过氧化氢酶(catalase,CAT)的活性。采用线性回归分析细胞内ROS的水平与ATP含量的相关性。结果LPF可以显著改善酒精诱导的线粒体能量代谢异常,增加线粒体呼吸链复合物的酶活性,促进细胞产生ATP,恢复线粒体膜电位。LPF能够缓解酒精诱导的氧化应激,降低细胞内ROS与MDA的水平,增强SOD与CAT的活性,增加GSH的水平。此外,LPF对ROS的抑制作用与对ATP的促进作用具有相关性(r=0.7595,P<0.0001)。结论LPF对酒精引起的线粒体损伤和能量代谢障碍具有改善作用,其机制与抑制氧化应激有关。OBJECTIVE To investigate the effects of corn peptide Leu-Pro-Phe(LPF)on ethanol-induced mitochondrial energy metabolism impairment and oxidative stress in C2C12 cells.METHODS The C2C12 cells were were induced to differentiate into muscle ducts and treated with ethanol(300 mmol·L^(-1))for 6 hours to establish ethanol stress model.The potential cellular protective effects of LPF(10,20,40μmol·L^(-1))werer evaluated based on this model.The levels of ATP and the activities of five respiratory chain complexes were measured using the corresponding assay kits.Rhodamine 123 staining was performed to detect mitochondrial membrane potential(MMP).H_2DCFDA probe was used to examine intracellular reactive oxygen species(ROS).Commercial assay kits were utilized to detect the contents of malondialdehyde(MDA)and glutathione(GSH),as well as the activities of superoxide dismutase(SOD)and catalase(CAT).Linear regression analysis was conducted to analyze the correlation between intracellular ROS levels and ATP content.RESULTS LPF significantly improved ethanol-induced mitochondrial energy metabolism abnormalities,increased the activities of mitochondrial respiratory chain complexes,promoted ATP production,and restored MMP.LPF alleviated ethanol-induced oxidative stress by reducing intracellular ROS and MDA,enhancing SOD and CAT activities,and increasing GSH.Furthermore,there showed significant correlation between the inhibitory effect of LPF on ROS and its promoting effect on ATP(r=0.7595,P<0.0001).CONCLUSION LPF can alleviate ethanol-induced mitochondrial damage and energy metabolism impairment,which may be related to its inhibition of oxidative stress.

关 键 词：玉米肽 Leu-Pro-Phe 酒精 氧化应激 线粒体 能量代谢 

分 类 号：R966[医药卫生—药理学]