CEDOC, Chronic Diseases Research Centre, NOVA Medical School/Faculdade de Ciência Médicas, Universidade Nova de Lisboa, 1169-056 Lisbon, Portugal.
Instituto de Biologia Experimental e Tecnológica (iBET), Apartado 12, 2781-901 Oeiras, Portugal.
FEMS Yeast Res. 2019 Aug 1;19(5). doi: 10.1093/femsyr/foz051.
Autophagy is an autodigestive process, promoting cytoprotection by the elimination of dysfunctional organelles, misfolded proteins and toxic aggregates. Carbon monoxide (CO) is an endogenous gasotransmitter that under low concentrations prevents cell death and inflammation. For the first time, the role of autophagy in CO-mediated cytoprotection against oxidative stress was evaluated in the model yeast Saccharomyces cerevisiae. The boron-based CO-releasing molecule, CORM-A1, was used to deliver CO. CORM-A1 partially prevented oxidative stress-induced cell death in yeast. Likewise, CORM-A1 activated autophagy under basal physiological conditions, which were assessed by autophagic flux and the expression of mCherry-Atg8 or GFP-Atg8. Inhibition of autophagy by knocking out key autophagic genes in yeast (ATG8 or ATG11) blocked CORM-A1 cytoprotective effect, indicating the critical role of autophagy in CO-induced cytoprotection. The CO-mediated cytoprotection via autophagy induction observed in yeast was validated in primary cultures of astrocytes, a well-characterized model for CO's cytoprotective functions. As in yeast, CORM-A1 prevented oxidative stress-induced cell death in an autophagy-dependent manner in astrocytes. Overall, our data support the cytoprotective action of CO against oxidative stress. CO promotes cytoprotection in yeast via autophagy, opening new possibilities for the study of molecular mechanisms of CO's biological functions using this powerful eukaryotic model.
自噬是一种自我消化的过程，通过消除功能失调的细胞器、错误折叠的蛋白质和有毒聚集体来促进细胞保护。一氧化碳（CO）是一种内源性气体递质，在低浓度下可防止细胞死亡和炎症。本文首次评估了自噬在 CO 介导的酵母 Saccharomyces cerevisiae 氧化应激细胞保护中的作用。使用基于硼的 CO 释放分子 CORM-A1 来输送 CO。CORM-A1 部分预防了酵母中氧化应激诱导的细胞死亡。同样，CORM-A1 在基础生理条件下激活自噬，通过自噬流和 mCherry-Atg8 或 GFP-Atg8 的表达来评估。敲除酵母中关键自噬基因（ATG8 或 ATG11）抑制自噬会阻断 CORM-A1 的细胞保护作用，表明自噬在 CO 诱导的细胞保护中起关键作用。在星形胶质细胞的原代培养中验证了 CO 通过诱导自噬介导的细胞保护，星形胶质细胞是 CO 细胞保护功能的一个很好的模型。与酵母一样，CORM-A1 以依赖自噬的方式防止氧化应激诱导的星形胶质细胞死亡。总之，我们的数据支持 CO 对抗氧化应激的细胞保护作用。CO 通过自噬促进酵母的细胞保护，为使用这种强大的真核模型研究 CO 生物学功能的分子机制开辟了新的可能性。
