[1]王雪梅 刘芬 曹莹 邢远 汪洁英 潘涛.HIF-1α介导线粒体功能调控糖尿病心肌缺血再灌注损伤的作用机制研究[J].心血管病学进展,2020,(10):1085.[doi:10.16806/j.cnki.issn.1004-3934.2020.10.021]
　WANG Xuemei,LIU Fen,CAO Ying,et al.HIF-1α Mediates the Regulation of Mitochondrial Function on Diabetic Myocardial Ischemia Reperfusion Injury[J].Advances in Cardiovascular Diseases,2020,(10):1085.[doi:10.16806/j.cnki.issn.1004-3934.2020.10.021]
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HIF-1α介导线粒体功能调控糖尿病心肌缺血再灌注损伤的作用机制研究()
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《心血管病学进展》[ISSN:51-1187/R/CN:1004-3934]

卷:
期数:
2020年10期
页码:
1085
栏目:
论著
出版日期:
2020-10-25

文章信息/Info

Title:
HIF-1α Mediates the Regulation of Mitochondrial Function on Diabetic Myocardial Ischemia Reperfusion Injury
作者:
王雪梅1; 2 刘芬2 曹莹1 邢远1 汪洁英1 潘涛1
 (1.西安医学院公共卫生学院,陕西 西安 710021;2.新疆医科大学第一附属医院动物疾病模型研究重点实验室,新疆 乌鲁木齐 830054)
Author(s):
WANG Xuemei1; 2;  LIU Fen2;  CAO Ying1;  XING Yuan1; WANG Jieying1;  PAN Tao1
(1.Department of public health,Xi’an Medical University,Xi’an 710021,Shaanxi,China; 2.Xinjiang Key Laboratory of Medical animal Model Research,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China)
关键词:
糖尿病心肌; 缺血再灌注; 低氧诱导因子1α; 过氧化物酶体增殖物激活受体γ辅助激活因子1α
Keywords:
Diabetic myocardium;  Ischemia/reperfusion;  Hypoxia-inducible factors 1α;  peroxisome proliferator activated receptor γ coactivator 1α
DOI:
10.16806/j.cnki.issn.1004-3934.2020.10.021
摘要:
目的 研究低氧诱导因子1α（HIF-1α）对高糖作用下缺血再灌注（IR）损伤的心肌细胞凋亡和氧化应激，及其信号通路下游因子的调控作用。方法 二甲基已二酰基甘氨酸激活HIF-1α对高糖作用下IR心肌细胞损伤的干预。实验分四组：对照组（细胞常规培养）、高糖+缺氧复氧组（25 mmol/L葡萄糖的DMEM培养基培养24 h，缺氧2 h，复氧8 h）、HIF-1α处理组（高糖，缺氧加复氧，缺氧前3 h用100 ?mol/L的二甲基已二酰基甘氨酸处理）、二甲基亚砜（DMSO）对照组（高糖，缺氧加复氧，缺氧前3 h用100 ?mol/L的DMSO处理）。流式细胞术测定细胞凋亡率，CCK-8法分析细胞活性；ELISA法检测细胞培养上清液中超氧化物歧化酶活性和丙二醛的表达；ATP含量试剂盒检测各组细胞ATP含量；Western Blot法检测过氧化物酶体增殖物激活受体γ辅助激活因子1α（PGC-1α）、核呼吸因子1，以及能量代谢相关的p-AMPK、AMPK蛋白表达，HIF-1α核蛋白表达。结果 高糖合并IR损伤增加心肌细胞的凋亡（P＜0.01），降低心肌细胞活力（P＜0.01），HIF-1α表达激活抑制心肌细胞的凋亡（P＜0.05）和活力降低（P＜0.05）。高糖合并IR损伤增加心肌细胞的氧化应激水平，HIF-1α表达激活抑制心肌细胞的氧化应激。和DMSO对照组相比，HIF-1α处理组的超氧化物歧化酶的表达显著增加[（11.56±0.72）U/mL vs（7.79±0.90）U/mL，P＜0.05]，丙二醛的表达显著降低[（0.69±0.07）nmol/mL vs（0.96±0.11）nmol/mL，P＜0.001]。高糖+缺氧复氧的应激降低了细胞的ATP含量，HIF-1α表达激活减轻了这种损伤。和DMSO对照组相比，HIF-1α处理组的ATP含量显著增加[（5.41±0.23）μmol/g vs（3.10±0.42）μmol/g，P＜0.05]。低氧条件下，HIF-1α核蛋白表达增加（P＜0.001），低氧激活p-AMPK蛋白表达（P＜0.01），线粒体氧化磷酸化的激活因子PGC-1α（P＜0.01）、转录因子核呼吸因子1（P＜0.01）的表达均显著上调。和DMSO组相比，HIF-1α处理组HIF-1α的表达显著增加（P＜0.05）、PGC-1α表达显著增加（P＜0.05）。结论 HIF-1α在糖尿病心肌IR损伤中通过提高PGC-1α的表达，提高线粒体的能量代谢水平，降低心肌的氧化应激损伤和凋亡。
Abstract:
Objective To study the effects of hypoxia inducible factor-1 α (HIF-1 α) on apoptosis and oxidative stress of myocardial cells induced by ischemia reperfusion injury under high glucose condition, and the downstream factors of the signaling pathway. Methods The intervention of HIF-1α was active d by d imethyloxallyl glycine (DMOG) on myocardial cell injury induced by high glucose and ischemia reperfusion. The experiment was divided into four groups:control group (conventional cell culture),high glucose+hypoxia reoxygenation group (DMEM medium culture of 25 mmol/L glucose for 24 h,hypoxia for 2 h,reoxygenation for 8 h),HIF-1α treatment group (high glucose,hypoxia reoxygenation,DMOG treatment of 100 μmol/L for 3 h before hypoxia), Dimethyl sulfoxide (DMSO) control group (high glucose ,hypoxia reoxygenation,DMSO treatment of 100 μmol/L for 3 h before hypoxia). Flow cytometry was used to detect the apoptosis rate,CCK-8 was used to analyze the cell activity,ELISA was used to detect the activity of SOD and the expression of MDA in the supernatant of cell culture, ATP content kit was used to detect the content of ATP in each group,Western Blot detected the protein expression of mitochondrial oxidative phosphorylation activator PGC-1α, transcription factor NRF-1, energy metabolism related factor AMPK,p-AMPK and HIF-1α nuclear proteins.Results The high glucose with ischemia reperfusion injury increase d the apoptosis of cardiomyocytes (P＜0.01),and decreased the viability of cardiomyocytes (P＜0.01). Activation of HIF-1α expression inhibited the apoptosis of cardiomyocytes(P < 0.05) and the decrease of the viability of cardiomyocytes ( P < 0.05) . The high glucose with ischemia reperfusion injury increased the oxidative stress of cardiomyocytes,and the activation of HIF-1α inhibited the oxidative stress of cardiomyocytes. Compared with DMSO control group ,the activity of SOD in HIF-1α treatment group increased significantly [(11.56±0.72)U/mL vs ( 7.79±0.90)U/mL,P＜0.05],and the expression of MDA decreased significantly[(0.69±0.07)nmol/mL vs ( 0.96±0.11)nmol/mL,P＜0.001]. The stress of high glucose+hypoxia reoxygenation reduced the ATP content of cells ,and HIF-1α relieved the injury. Compared with DMSO control group,ATP content of HIF-1α treatment group increased significantly[(5.41±0.23)μmol/g vs ( 3.10±0.42)μmol/g,P＜0.05]. Under hypoxia,the expression of HIF-1α nuclear protein increased (P＜0.001),the expression of p-AMPK protein increased (P＜0.01),the expression of mitochondrial oxidative phosphorylation activator PGC-1α increased (P＜0.01),and transcription factor NRF-1 increased (P＜0.01). Compared with DMSO group,the expression of HIF-1α and PGC-1α in HIF-1α treatment group increased significantly (P＜0.05).Conclusion HIF-1α can increase the expression of PGC-1α,improve the energy metabolism level of mitochondria,and reduce the oxidative stress injury and apoptosis of myocardium

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更新日期/Last Update: 2020-12-21