Recombinant Atrial Natriuretic Peptide Prevents Aberrant Ca2+ Leakage through the Ryanodine Receptor by Suppressing Mitochondrial Reactive Oxygen Species Production Induced by Isoproterenol in Failing Cardiomyocytes.
Recombinant Atrial Natriuretic Peptide Prevents Aberrant Ca2+ Leakage through the Ryanodine Receptor by Suppressing Mitochondrial Reactive Oxygen Species Production Induced by Isoproterenol in Failing Cardiomyocytes.
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DOI：
10.1371/journal.pone.0163250
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Yano M
中科院分区：
文献类型：
--
作者：
Murakami W;Kobayashi S;Susa T;Nanno T;Ishiguchi H;Myoren T;Nishimura S;Kato T;Hino A;Oda T;Okuda S;Yamamoto T;Yano M
儿茶酚胺诱导细胞内的ROS，从而在心力衰竭(HF)时通过兰尼定受体增加舒张期的钙泄漏。然而，心钠素(ANP)对衰竭心肌细胞ROS生成和钙处理的影响知之甚少。本研究的目的是阐明外源性心钠素在心力衰竭期间发挥心脏保护作用的机制。从心动过速诱导的心衰犬模型和假手术对照组的左心室分离心肌细胞。用抗VDAC抗体和8-羟基-2‘-脱氧鸟苷抗体进行双重免疫组织化学(IHC)染色，评价线粒体DNA氧化程度。用2，7-二氯二乙酸酯荧光素观察ANP对心肌细胞ROS的影响，用荧光素-4-AM共聚焦显微镜观察心肌细胞舒张期钙火花的变化，以及48h后心肌细胞存活率的变化。双重免疫组化结果显示，异丙肾上腺素(ISO)可显著增加HF大鼠心肌线粒体氧化DNA的含量，同时ANP可显著抑制ISO引起的心肌细胞DNA损伤。与正常对照组相比，心力衰竭患者ROS生成和钙触发频率(CASF)均显著增加，且在ISO存在的情况下进一步增加。值得注意的是，ANP显著抑制ISO诱导的ROS和CASF，而不改变HF的肌浆网钙含量(p<；0.01)。与基线相比，在ISO存在的情况下，在HF中48h的存活率显著降低(p<；0.01)，而在ANP的共同存在下，存活率显著提高(p<；0.01)。总之，我们的结果表明，ANP强烈抑制ISO诱导的线粒体ROS的产生，这可能纠正了舒张期钙离子的异常火花，最终有助于提高心衰心肌细胞的存活率。
Catecholamines induce intracellular reactive oxygen species (ROS), thus enhancing diastolic Ca2+ leakage through the ryanodine receptor during heart failure (HF). However, little is known regarding the effect of atrial natriuretic peptide (ANP) on ROS generation and Ca2+ handling in failing cardiomyocytes. The aim of the present study was to clarify the mechanism by which an exogenous ANP exerts cardioprotective effects during HF. Cardiomyocytes were isolated from the left ventricles of a canine tachycardia-induced HF model and sham-operated vehicle controls. The degree of mitochondrial oxidized DNA was evaluated by double immunohistochemical (IHC) staining using an anti-VDAC antibody for the mitochondria and an anti-8-hydroxy-2′-deoxyguanosine antibody for oxidized DNA. The effect of ANP on ROS was investigated using 2,7-dichlorofluorescin diacetate, diastolic Ca2+ sparks assessed by confocal microscopy using Fluo 4-AM, and the survival rate of myocytes after 48 h. The double IHC study revealed that isoproterenol (ISO) markedly increased oxidized DNA in the mitochondria in HF and that the ISO-induced DNA damage was markedly inhibited by the co-presence of ANP. ROS production and Ca2+ spark frequency (CaSF) were increased in HF compared to normal controls, and were further increased in the presence of ISO. Notably, ANP significantly suppressed both ISO-induced ROS and CaSF without changing sarcoplasmic reticulum Ca2+ content in HF (p<0.01, respectively). The survival rate after 48 h in HF was significantly decreased in the presence of ISO compared with baseline (p<0.01), whereas it was significantly improved by the co-presence of ANP (p<0.01). Together, our results suggest that ANP strongly suppresses ISO-induced mitochondrial ROS generation, which might correct aberrant diastolic Ca2+ sparks, eventually contributing to the improvement of cardiomyocyte survival in HF.
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影响因子：
20.1
作者：
Luo M;Anderson ME
通讯作者：
Anderson ME
影响因子：
3.7
作者：
Kobayashi S;Susa T;Ishiguchi H;Myoren T;Murakami W;Kato T;Fukuda M;Hino A;Suetomi T;Ono M;Uchinoumi H;Tateishi H;Mochizuki M;Oda T;Okuda S;Doi M;Yamamoto T;Yano M
通讯作者：
Yano M
影响因子：
3.4
作者：
Bovo, Elisa;Mazurek, Stefan R.;Zima, Aleksey V.
通讯作者：
Zima, Aleksey V.
DOI：
10.1073/pnas.1103300108
发表时间：
2011-11-08
影响因子：
11.1
作者：
Klaiber, Michael;Dankworth, Beatrice;Kuhn, Michaela
通讯作者：
Kuhn, Michaela
DOI：
10.1016/s0735-1097(01)01233-5
发表时间：
2001-06-01
影响因子：
24
作者：
Hayashi, M;Tsutamoto, T;Kinoshita, M
通讯作者：
Kinoshita, M