Endogenous H2S resists mitochondria-mediated apoptosis in the adrenal glands via ATP5A1 S-sulfhydration in male mice
Endogenous H2S resists mitochondria-mediated apoptosis in the adrenal glands via ATP5A1 S-sulfhydration in male mice
复制标题
内源性 H 2 S 通过 ATP5A1 S-硫化作用抵抗雄性小鼠肾上腺中线粒体介导的细胞凋亡
DOI：
10.1016/j.mce.2018.02.011
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发表时间：
2018-10-15
影响因子：
4.1
通讯作者：
Ni, Xin
中科院分区：
文献类型：
--
作者：
Wang, Changnan;Du, Jiankui;Ni, Xin
在先前的研究中，我们发现内源性硫化氢（H2S）通过保护线粒体功能在维持完整的肾上腺皮质功能中起着关键作用。我们进一步研究了H2S对肾上腺功能的保护作用是否与H2S介导的细胞凋亡有关。LPS处理导致雄性小鼠肾上腺中的细胞凋亡，并且这些作用被H2S供体GYY 4137阻止。在Y1细胞模型中，GYY 4137可挽救LPS诱导的细胞凋亡和对ACTH的钝性反应。H2S生成酶胱硫醚-β-合酶（CBS）敲除杂合（CBS+/-）小鼠显示肾上腺中的细胞凋亡和肾上腺功能不全。GYY 4137治疗恢复了肾上腺功能并消除了肾上腺介导的细胞凋亡。马来酰亚胺法结合质谱分析表明，肾上腺线粒体中的一些蛋白质是S-巯基化的。使用改良的生物素开关测定法进一步证实ATP 5A 1为S-巯基化的。内毒素血症和CBS+/-小鼠肾上腺中S-硫代ATP 5A 1的水平降低，GYY 4137可恢复该水平。ATP 5A 1被鉴定为通过H2S在半胱氨酸244处硫氢化。在Y1细胞中过表达半胱氨酸244突变体ATP 5A 1导致LPS诱导的细胞凋亡的丧失和GYY 4137恢复LPS诱导的ACTH低反应性。总的来说，本研究表明，减少H2S的产生导致肾上腺皮质细胞介导的细胞凋亡和对ACTH的钝性反应。ATP 5A 1在半胱氨酸244处的S-巯基化是H2S维持肾上腺中线粒体功能和类固醇生成的重要分子机制。(C)2018 Elsevier B. V.版权所有。
In a previous study, we showed that endogenous hydrogen sulfide (H2S) plays a key role in the maintenance of intact adrenal cortex function via the protection of mitochondrial function during endoxemia. We further investigated whether mitochondria-mediated apoptosis is involved in H2S protection of adrenal function. LPS treatment resulted in mitochondria-mediated apoptosis in the adrenal glands of male mice, and these effects were prevented by the H2S donor GYY4137. In the model of Y1 cells, the LPS-induced mitochondria-mediated apoptosis and blunt response to ACTH were rescued by GYY4137. The H2S-generating enzyme cystathionine-beta-synthase (CBS) knockout heterozygous (CBS+/-) mice showed mitochondria-mediated apoptosis in the adrenal gland and adrenal insufficiency. GYY4137 treatment restored adrenal function and eliminated mitochondria-mediated apoptosis. Maleimide assay combined with mass spectrometry analysis showed that a number of proteins in mitochondria were S-sulfhydrated in the adrenal gland. ATP5A1 was further confirmed as S-sulfhydrated using a modified biotin switch assay. The level of S-sulfhydrated ATP5A1 was decreased in the adrenal gland of endotoxemic and CBS+/- mice, which was restored by GYY4137. ATP5A1 was identified as sulfhydrated at cysteine 244 by H2S. Overexpression of the cysteine 244 mutant ATP5A1 in Y1 cells resulted in a loss of LPS-induced mitochondria-mediated apoptosis and GYY4137 restoration of LPS-induced hyporesponsiveness to ACTH. Collectively, the present study revealed that decreased H2S generation leads to mitochondrial-mediated apoptosis in the adrenal cortex and a blunt response to ACTH. S-sulfhydration of ATP5A1 at cysteine 244 is an important molecular mechanism by which H2S maintains mitochondrial function and steroidogenesis in the adrenal glands. (C) 2018 Elsevier B.V. All rights reserved.