解偶联蛋白2过表达对脓毒症大鼠心肌细胞线粒体的保护作用

doi:10.13481/j.1671-587x.20180402

Abstract

Abstract: Objective:To explore the protective effect of overexpression of uncoupling protein 2 (UCP2) on the mitochondrial in cardiomyocytes of the rats with sepsis, and to clarify its mechanisms. Methods:The H9C2 cardiomyocytes with UCP2 overexpression were reconstructed and randomly devided into normal control group, lipopolysaccharide/peptidoglycan(LPS/PepG) group,PHBLV group and PHBLV-UCP2 group.The H9C2 cells in various groups except normal control group were stimulated with LPS/PepG.The expression levels of UCP2 mRNA and protein in the cardiomyocytes in various groups were detected with RT-PCR and Western blotting methods.The levels of creatine kinase (CK) and tumor necrosis factorα(TNF-α) were measured by spectrophotometer and enzyme-linked immunosorbent assay (ELISA).The mitochondrial morphology of cardiomyocytes in various groups was observed by transmission electron microscope (TEM),the mitochondrial membrane potential (MMP) levels of cardiomyocytes in various groups were detected by flow cytometry (FCM),and the levels of reactive oxygen species (ROS) and adenosine triphosphate (ATP) in mitochondria of cardiomyocytes were measured by multimode reader. Results:Compared with normal control group,the levels of CK and TNF-α in LPS/PepG group were increased significantly(P<0.05),the mitochondrial morphology was injured obviously,the MMP level was decreased (P<0.05),the level of ROS was increased (P<0.05), and the ATP level was decreased (P<0.05).Compared with LPS/PepG group,the CK and TNF-α levels in cardiomyocytes in PHBLV-UCP2 group were decreased (P<0.05),the injury of mitochondria was improved,the MMP level was increased (P<0.05),the ROS level was decreased signficantly(P<0.05),and the ATP level was significantly increased(P<0.05).The levels of CK,TNF-α,MMP,ROS,ATP in the cardiomyocytes and the degree of mitochondrial swelling in PHBLV group had no obovionsly changes compared with LPS/PepG group (P>0.05). Conclusion: UCP2 overexpression plays a protective role in myocardial mitochondria of the rats with sepsis,which may be related to its regulation on the membrane potential to affect the mitochondrial ROS and ATP synthesis through uncoupling effect under septic condiction.

Key words: uncoupling protein 2, sepsis, mitochondria, cardiomyocytes

CLC Number:

R554.8

ZHENG Guilang, GUO Yuxiong, LYU Juanjuan, HUANG Jinda, LIU Cui, ZENG Qiyi. Protective effect of overexpression of uncoupling protein 2 on mitochondria in cardiomyocytes of rats with sepsis[J].Journal of Jilin University Medicine Edition, 2018, 44(04): 698-703.

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Protective effect of overexpression of uncoupling protein 2 on mitochondria in cardiomyocytes of rats with sepsis

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