Evaluation of Parkin in the Regulation of Myocardial Mitochondria-Associated Membranes and Cardiomyopathy During Endotoxemia.
Evaluation of Parkin in the Regulation of Myocardial Mitochondria-Associated Membranes and Cardiomyopathy During Endotoxemia.
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评价Parkin对内毒素血症心肌线粒体相关膜和心肌病的调节作用。
DOI：
10.3389/fcell.2022.796061
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发表时间：
2022
影响因子：
5.5
通讯作者：
Zang QS
中科院分区：
文献类型：
--
作者：
Kim M;Nikouee A;Sun Y;Zhang QJ;Liu ZP;Zang QS
背景：线粒体缺陷是脓毒症引起的器官衰竭的一种已知病理。我们以前发现，在心脏内毒素血症期间，支持线粒体状态的亚细胞结构域-通过E3泛素连接酶Parkin和PTEN诱导的激酶1（PINK 1）的线粒体自噬途径控制线粒体质量。这里描述的研究检查了帕金对心脏MAMs和内毒素血症诱导的心肌病的影响。此外，Parkin的点突变W 403 A先前被鉴定为体外组成型活性突变。在内毒素血症模型中评价了该突变的强制表达的体内效应。 研究方法：对野生型（WT）、Parkin缺陷型（Park 2-/-）和Parkin W 402 A（人Parkin W 403 A）敲入表达的小鼠进行脂多糖（LPS）攻击。超声心动图评价心功能。在收获的心脏组织中，通过超离心分离MAM组分，并定量其量和功能。电镜观察MAMs和线粒体的超微结构。线粒体呼吸活性测定酶法。通过促炎细胞因子IL-6的水平来估计心肌炎症。通过与线粒体相关的线粒体自噬因子水平和线粒体-溶酶体共定位程度来评估心肌线粒体自噬。还评价了帕金激活，其通过帕金的丝氨酸65上的磷酸化来表示。 结果如下：与WT相比，Park 2 −/−小鼠在内毒素血症期间表现出更严重的心脏MAMs受损，其特征在于结构破坏，数量减少和转运功能减弱。内毒素血症诱导的心肌病在Park 2 −/−小鼠中加剧，表现为心肌收缩力恶化和IL-6产生增加。来自Park 2 −/−心脏的线粒体更加恶化，表现为结构完整性和呼吸功能的丧失。出乎意料的是，携带Parkin W 402 A的小鼠与WT小鼠相比，表现出相似的心肌病和线粒体损伤水平。此外，Parkin W 402 A突变既不增强线粒体自噬，也不增加内毒素血症的挑战下心肌中的Parkin激活。 结论：Parkin/PINK 1线粒体自噬参与了内毒素血症时心肌MAMs的调节。Parkin基因点突变W 402 A（人W 403 A）不足以减轻体内内毒素血症引起的心肌病。
Background: Mitochondrial deficiency is a known pathology in sepsis-induced organ failure. We previously found that mitochondria-associated membranes (MAMs), a subcellular domain supporting mitochondrial status, are impaired in the heart during endotoxemia, suggesting a mechanism of mitochondrial damage occurred in sepsis. Mitophagy pathway via E3 ubiquitin ligase Parkin and PTEN-induced kinase 1 (PINK1) controls mitochondrial quality. Studies described here examined the impact of Parkin on cardiac MAMs and endotoxemia-induced cardiomyopathy. Additionally, point mutation W403A in Parkin was previously identified as a constitutively active mutation in vitro. In vivo effects of forced expression of this mutation were evaluated in the endotoxemia model. Methods: Mice of wild type (WT), Parkin-deficiency (Park2 −/− ), and knock-in expression of Parkin W402A (human Parkin W403A) were given lipopolysaccharide (LPS) challenge. Cardiac function was evaluated by echocardiography. In the harvested heart tissue, MAM fractions were isolated by ultracentrifugation, and their amount and function were quantified. Ultrastructure of MAMs and mitochondria was examined by electron microscopy. Mitochondrial respiratory activities were measured by enzyme assays. Myocardial inflammation was estimated by levels of pro-inflammatory cytokine IL-6. Myocardial mitophagy was assessed by levels of mitophagy factors associated with mitochondria and degrees of mitochondria-lysosome co-localization. Parkin activation, signified by phosphorylation on serine 65 of Parkin, was also evaluated. Results: Compared with WT, Park2 −/− mice showed more severely impaired cardiac MAMs during endotoxemia, characterized by disrupted structure, reduced quantity, and weakened transporting function. Endotoxemia-induced cardiomyopathy was intensified in Park2 −/− mice, shown by worsened cardiac contractility and higher production of IL-6. Mitochondria from the Park2 −/− hearts were more deteriorated, indicated by losses in both structural integrity and respiration function. Unexpectedly, mice carrying Parkin W402A showed similar levels of cardiomyopathy and mitochondrial damage when compared with their WT counterparts. Further, Parkin W402A mutation neither enhanced mitophagy nor increased Parkin activation in myocardium under the challenge of endotoxemia. Conclusion: our results suggest that Parkin/PINK1 mitophagy participates in the regulation of cardiac MAMs during endotoxemia. Point mutation W402A (human W403A) in Parkin is not sufficient to alleviate cardiomyopathy induced by endotoxemia in vivo.
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影响因子：
11.4
作者：
Area-Gomez, Estela;Castillo, Maria Del Carmen Lara;Tambini, Marc D.;Guardia-Laguarta, Cristina;de Groof, Ad J. C.;Madra, Moneek;Ikenouchi, Junichi;Umeda, Masato;Bird, Thomas D.;Sturley, Stephen L.;Schon, Eric A.
通讯作者：
Schon, Eric A.
影响因子：
12.4
作者：
Michiorri, S.;Gelmetti, V.;Casari, G.
通讯作者：
Casari, G.
影响因子：
8.8
作者：
Levy, Mitchell M.;Dellinger, R. Phillip;Angus, Derek C.
通讯作者：
Angus, Derek C.
影响因子：
13.3
作者：
Jin, Seok Min;Youle, Richard J.
通讯作者：
Youle, Richard J.
影响因子：
13.3
作者：
Gelmetti, Vania;De Rosa, Priscilla;Valente, Enza Maria
通讯作者：
Valente, Enza Maria