| 摘要: |
| 摘要：目的：探究NOTCH2对急性心肌梗死（AMI）的影响及其具体分子机制。
方法：使用左前降之冠状动脉结扎法构建大鼠AMI模型，使用氧糖剥夺法构建心肌细胞体外模型。分别使用2,3,5-氯化三苯基四氮唑（TTC）染色和苏木素伊红（HE）染色观察大鼠心肌缺血情况和大鼠心肌组织形态。使用细胞计数试剂盒8（CCK-8）检测心肌细胞活性；使用流式细胞术检测心肌细胞凋亡情况；使用免疫荧光检测心肌细胞中NOTCH2表达情况。使用蛋白免疫印迹（WB）检测组织和细胞中PKLR、p-PKM2、PKM2、NOTCH2、Wnt3a和β-catenin蛋白表达情况。
结果：与Sham组大鼠相比，AMI组大鼠发生了明显的心肌缺血和心肌损伤，其心肌组织中NOTCH2表达上调，糖酵解水平下调，Wnt/β-catenin信号通路异常激活。与Control组细胞相比，氧糖剥夺组心肌细胞活性下降，凋亡水平上升，NOTCH2表达异常升高，糖酵解水平下调，Wnt/β-catenin信号通路异常激活。干扰NOTCH2表达后，心肌细胞活性上升，凋亡水平下降，糖酵解水平上升，Wnt/β-catenin信号通路被抑制。另外，糖酵解抑制剂2-DG和Wnt/β-catenin信号通路激动剂SKL2001的使用能显著回复NOTCH2低表达对心肌细胞的影响。 |
| 关键词: 急性心肌梗死 糖代谢重编程 NOTCH2 Wnt/β-catenin信号通路 |
| DOI： |
| 分类号: |
| 基金项目:浙江省医药卫生科技计划项目 |
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| The mechanism of activation of Wnt/β-catenin signaling pathway by NOTCH2 to drive glucose metabolic reprogramming and promote acute myocardial infarction |
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maxuhui1, cen mingqiu2, luming1, yujia1
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1.Hangzhou Xixi Hospital;2.xixi hospital of Hangzhou
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| Abstract: |
| Abstract:
Objective: To explore the effect of NOTCH2 on acute myocardial infarction (AMI) and its molecular mechanism.
Methods: Rat AMI model was constructed by left anterior descending coronary artery ligation and myocardial cell model was constructed by oxygen-glucose deprivation. 2,3, 5-triphenyltetrazolium chloride (TTC) staining and hematoxylin eosin (HE) staining were used to observe myocardial ischemia and myocardial tissue morphology, respectively. Cell counting kit 8 (CCK-8) was used to detect cardiomyocyte activity. The apoptosis of cardiomyocytes was detected by flow cytometry. The expression of NOTCH2 in cardiomyocytes was detected by immunofluorescence. Protein expression in tissues and cells was detected by Western blot (WB).
Results: Compared with rats in Sham group, rats in AMI group had obvious myocardial ischemia and myocardial injury, NOTCH2 expression was up-regulated, glycolysis level was down-regulated, and Wnt/β-catenin signaling pathway was abnormally activated. Compared with the Control group, the activity of cardiomyocytes in OGD group decreased, the apoptosis level increased, the expression of NOTCH2 was abnormally increased, the glycolysis level was down-regulated, and the Wnt/β-catenin signaling pathway was abnormally activated. After interfering NOTCH2 expression, the activity of cardiomyocytes increased, the apoptosis level decreased, the glycolysis level increased, the glycolysis level decreased, and the Wnt/β-catenin signaling pathway was inhibited. In addition, the use of glycolytic inhibitor 2-DG and Wnt/β-catenin signaling pathway agonist SKL2001 can significantly recover the effects of NOTCH2 low expression on cardiomyocytes.
Conclusion: NOTCH2 is highly expressed in AMI. It was further proved that NOTCH2 may promote the malignant progression of AMI by inhibiting glycolysis by activating Wnt/β-catenin signaling pathway. |
| Key words: Acute myocardial infarction Glucose metabolism reprogramming glycolysis NOTCH2 Wnt/β-catenin signaling pathway |