Necroptosis in primate luteolysis: a role for ceramide
Necroptosis in primate luteolysis: a role for ceramide
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DOI：
10.1038/s41420-019-0149-7
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发表时间：
2019-02-11
影响因子：
7
通讯作者：
Mayerhofer, Artur
中科院分区：
文献类型：
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作者：
Bagnjuk, Konstantin;Stoeckl, Jan Bernd;Mayerhofer, Artur
黄体（CL）是一个短暂的内分泌器官，但导致其死亡的分子机制还不清楚。人和非人灵长类CL样本（猕猴和猕猴）中存在磷酸化混合谱系激酶结构域样假激酶pMLKL（T357/S358），这意味着可能涉及黄体细胞的坏死性凋亡。In M. mulatta CL，pMLKL阳性染色仅从黄体中晚期开始才可检测到，表明CL消退期间出现坏死性凋亡。细胞死亡，包括坏死性凋亡，先前在人黄体颗粒细胞（GC）的培养物中观察到，这是一种适合研究人CL的模型。为了探索培养过程中GC中坏死性凋亡细胞死亡的机制，我们进行了蛋白质组学分析。50种蛋白质的水平在培养5天后显著改变。互联分析和免疫细胞化学特别暗示神经酰胺补救途径得到加强。M. mulatta CL转录组分析表明体内相关性。伏马菌素B1（FB 1）和添加可溶性神经酰胺（C2-CER）干扰内源性神经酰胺产生对GC活力产生相反的作用，因此支持神经酰胺途径的重要性。形态学变化表明C2-CER处理组中的坏死细胞死亡。使用泛半胱天冬酶阻断剂zVAD-favor或坏死性凋亡阻断剂necrosulfonamid（NSA）的研究进一步支持C2-CER诱导坏死性凋亡。我们的数据指出了生理过程中的坏死性凋亡，即CL回归。这提出了灵长类动物CL可以通过药理学抑制坏死性凋亡或通过与神经酰胺代谢的相互作用来挽救的可能性。
The corpus luteum (CL) is a transient endocrine organ, yet molecular mechanisms resulting in its demise are not well known. The presence of phosphorylated mixed lineage kinase domain-like pseudokinase pMLKL(T357/S358) in human and nonhuman primate CL samples (Macaca mulatto and Callithrix jacchus) implied that necroptosis of luteal cells may be involved. In M. mulatta CL, pMLKL positive staining became detectable only from the mid-late luteal phase onwards, pointing to necroptosis during regression of the CL. Cell death, including necroptosis, was previously observed in cultures of human luteal granulosa cells (GCs), an apt model for the study of the human CL. To explore mechanisms of necroptotic cell death in GCs during culture, we performed a proteomic analysis. The levels of 50 proteins were significantly altered after 5 days of culture. Interconnectivity analysis and immunocytochemistry implicated specifically the ceramide salvage pathway to be enhanced. M. mulatta CL transcriptome analysis indicated in vivo relevance. Perturbing endogenous ceramide generation by fumonisin B1 (FB1) and addition of soluble ceramide (C2-CER) yielded opposite actions on viability of GCs and therefore supported the significance of the ceramide pathway. Morphological changes indicated necrotic cell death in the C2-CER treated group. Studies with the pan caspase blocker zVAD-fmk or the necroptosis blocker necrosulfonamid (NSA) further supported that C2-CER induced necroptosis. Our data pinpoint necroptosis in a physiological process, namely CL regression. This raises the possibility that the primate CL could be rescued by pharmacological inhibition of necroptosis or by interaction with ceramide metabolism.