NADPH抗心肌肥厚作用的机制研究
首发时间:2023-05-18
摘要:目的:在本研究中,我们主要对还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)抗心肌肥厚作用的机制进行研究。 方法:采用主动脉弓狭窄术(TAC)诱导小鼠病理性心肌肥厚及心衰模型。检测心重指数、左心室指数。采用血管紧张素II(AngII)构建体外H9C2心肌细胞肥大模型,免疫荧光a-actin染色估心肌细胞表面积。采用高通量呼吸检测仪Oxygraph-2k测量线粒体呼吸。采用qPCR检查心肌肥厚标志物ANP和BNP以及线粒体电子传递链复合物亚基细胞色素C氧化酶亚基1(MTCO1)和ATP合成酶F1亚基(ATP5A1)的mRNA表达水平,JC-1染色测定线粒体膜电位。Western blot检测去乙酰化酶3(SIRT3)、视神经萎缩蛋白1(OPA1)和线粒体融合蛋白2(MFN2)表达, 检测AMP依赖蛋白激酶(AMPK)和肝脏激酶B1(LKB1)的磷酸化水平。 结果:在体内压力超负荷诱导的心肌肥厚和心衰模型中,NADPH明显降低了经TAC诱导后的心重指数、左心室指数以及肥厚标志物ANP 和BNP的mRNA水平。NADPH提高了TAC诱导心肌肥厚小鼠的线粒体呼吸电子传递链复合物MTCO1和ATP5A1的mRNA水平,也增强了线粒体呼吸能力。在AngII诱导H9C2心肌细胞肥大模型中,NADPH降低了AngII诱导的ANP和BNP的mRNA水平和细胞表面积。NADPH明显改善AngII诱导的线粒体膜电位,增加L-OPA1和MFN2蛋白水平,维持线粒体融合裂变稳态。NADPH提高SIRT3蛋白表达, 激活LKB1-AMPK信号通路。 结论:我们的研究数据表明外源性NADPH具有抗心肌肥厚作用,其机制可能是NADPH激活SIRT3-LKB1-AMPK信号通路和改善线粒体功能。
关键词: NADPH 心肌肥厚 线粒体呼吸 线粒体动力学 SIRT3
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Study on the mechanism of anti-cardiac hypertrophy effects of NADPH
Abstract:Aim: In this study, we mainly studied the mechanism of anti-myocardial hypertrophy effect of reduced niacinamide adenine dinucleotide phosphate (NADPH). Methods: Transverse aortic constriction (TAC) was used to induce cardiac hypertrophy and heart failure in mice. Heart weight index, left ventricular index were detected. Angitensin II was used to induce H9C2 cell cardiac hypertrophy.The surface area of cardiomyocyte was evaluated by α-actin immunofluorescence staining. Mitochondrial respiration was measured using a high-throughput respiratory detector oxygraph-2k. The mRNA expression of ANP and BNP, and mitochondrial electron transport chain complex subunits cytochrome c oxidase subunit 1(MTCO1) and ATP synthase F1 subunit (ATP5A1) were detected by qPCR. Mitochondrial membrane potential was determined by JC-1 staining. The expression of sirtuin 3(SIRT3), optic atrophy protein 1 (OPA1), mitochondrial fusion protein 2(MFN 2), and the phosphorylation levels of AMP-dependent protein kinase (AMPK) and liver kinase B1(LKB1) were detected by Western blot. Results: In the in vivo model of pressure overload induced myocardial hypertrophy and heart failure, NADPH significantly reduced the TAC-induced increase of heart weight index, left ventricular index and mRNA levels of the hypertrophic marker ANP and BNP. NADPH increased the mRNA levels of MTCO1 and ATP5A1 and also enhanced mitochondrial respiratory capacity. In the AngII- induced in vitro H9c2 cardiomyocyte hypertrophy model, NADPH also reduced AngII-induced ANP and BNP mRNA levels and cell surface area. In addition, NADPH treatment significantly improved mitochondrial membrane potential, increased L-OPA1 and MFN2 protein levels to maintain the mitochondrial fusion and fission homeostasis. NADPH increased SIRT3 expression and activated LKB1-AMPK signaling. Conclusion: Our data suggest that exogenous NADPH may prevent cardiac hypertrophy by activating SIRT3-LKB1-AMPK signaling pathway and improving mitochondrial function.
Keywords: NADPH cardiac hypertrophy mitochondrial respiration mitochondrial dynamics SIRT3
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