circ-CBFB upregulates p66Shc to perturb mitochondrial dynamics in APAP-induced liver injury.
circ-CBFB upregulates p66Shc to perturb mitochondrial dynamics in APAP-induced liver injury.
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DOI：
10.1038/s41419-020-03160-y
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发表时间：
2020-11-06
影响因子：
9
通讯作者：
Yao J
中科院分区：
文献类型：
--
作者：
Wang Z;Zhao Y;Sun R;Sun Y;Liu D;Lin M;Chen Z;Zhou J;Lv L;Tian X;Yao J
p66Shc 是线粒体活性氧 (mtROS) 的主要调节因子，是肝细胞氧化应激的重要调节因子。然而，其对对乙酰氨基酚 (APAP) 诱导的肝损伤的功能贡献及其调节机制仍不清楚。在这里，我们的目的是评估 p66Shc 对 APAP 诱导的肝损伤的影响，并评估环状 RNA (circRNA) 是否作为竞争性内源性 RNA (ceRNA) 介导 p66Shc 在 APAP 诱导的肝损伤中。 p66Shc、miR-185-5p 和 circ-CBFB 沉默小鼠注射 APAP。在 APAP 刺激之前，用 p66Shc、miR-185-5p 和 circ-CBFB 沉默或过表达质粒或 siRNA 转染 AML12 细胞。 p66Shc 在 APAP 反应下在肝组织中上调，体内 p66Shc 沉默可以保护小鼠免受 APAP 诱导的线粒体动力学扰动和肝损伤。体外 p66Shc 敲低可减弱线粒体动力学和 APAP 诱导的肝细胞损伤。从机械角度来看，p66Shc 部分通过抑制 OMA1 泛素化来扰乱线粒体动力学。通过微阵列和生物信息学分析鉴定出直接抑制p66Shc翻译的miR-185-5p，其过度表达在体外减弱了线粒体动力学和肝细胞损伤。此外，荧光素酶、pull-down 和 RNA 免疫沉淀测定表明，circ-CBFB 作为 miR-185-5p 的 miRNA 海绵，在 APAP 诱导的肝损伤中介导 p66Shc。 circ-CBFB 敲低还减轻了 APAP 诱导的线粒体动力学扰动和肝细胞损伤。更重要的是，我们发现 circ-CBFB 敲低对 p66Shc、线粒体动力学和肝损伤的保护作用在体内和体外均被 miR-185-5p 抑制所消除。总之，p66Shc 是 APAP 诱导的肝损伤的关键调节因子，通过触发线粒体动力学扰动发挥作用。 circ-CBFB 在 APAP 诱导的肝损伤过程中作为 ceRNA 调节 p66Shc，这可能提供潜在的治疗靶点。
p66Shc, a master regulator of mitochondrial reactive oxygen species (mtROS), is a crucial mediator of hepatocyte oxidative stress. However, its functional contribution to acetaminophen (APAP)-induced liver injury and the mechanism by which it is modulated remain unknown. Here, we aimed to assess the effect of p66Shc on APAP-induced liver injury and to evaluate if circular RNA (circRNA) functions as a competitive endogenous RNA (ceRNA) to mediate p66Shc in APAP-induced liver injury. p66Shc-, miR-185-5p-, and circ-CBFB-silenced mice were injected with APAP. AML12 cells were transfected with p66Shc, miR-185-5p, and circ-CBFB silencing or overexpression plasmids or siRNAs prior to APAP stimulation. p66Shc was upregulated in liver tissues in response to APAP, and p66Shc silencing in vivo protected mice from APAP-induced mitochondrial dynamics perturbation and liver injury. p66Shc knockdown in vitro attenuated mitochondrial dynamics and APAP-induced hepatocyte injury. Mechanically, p66Shc perturbs mitochondrial dynamics partially by inhibiting OMA1 ubiquitination. miR-185-5p, which directly suppressed p66Shc translation, was identified by microarray and bioinformatics analyses, and its overexpression attenuated mitochondrial dynamics and hepatocyte injury in vitro. Furthermore, luciferase, pull-down and RNA immunoprecipitation assays demonstrated that circ-CBFB acts as a miRNA sponge of miR-185-5p to mediate p66Shc in APAP-induced liver injury. circ-CBFB knockdown also alleviated APAP-induced mitochondrial dynamics perturbation and hepatocyte injury. More importantly, we found that the protective effects of circ-CBFB knockdown on p66Shc, mitochondrial dynamics and liver injury were abolished by miR-185-5p inhibition both in vivo and in vitro. In conclusion, p66Shc is a key regulator of APAP-induced liver injury that acts by triggering mitochondrial dynamics perturbation. circ-CBFB functions as a ceRNA to regulate p66Shc during APAP-induced liver injury, which may provide a potential therapeutic target.
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影响因子：
64.5
作者：
Giorgio, M;Migliaccio, E;Pelicci, PG
通讯作者：
Pelicci, PG
影响因子：
4
作者：
Houghton, Benjamin C.;Booth, Claire;Thrasher, Adrian J.
通讯作者：
Thrasher, Adrian J.
影响因子：
120.7
作者：
Kris, Mark G.;Johnson, Bruce E.;Berry, Lynne D.;Kwiatkowski, David J.;Iafrate, A. John;Wistuba, Ignacio I.;Varella-Garcia, Marileila;Franklin, Wilbur A.;Aronson, Samuel L.;Su, Pei-Fang;Shyr, Yu;Camidge, D. Ross;Sequist, Lecia V.;Glisson, Bonnie S.;Khuri, Fadlo R.;Garon, Edward B.;Pao, William;Rudin, Charles;Schiller, Joan;Haura, Eric B.;Socinski, Mark;Shirai, Keisuke;Chen, Heidi;Giaccone, Giuseppe;Ladanyi, Marc;Kugler, Kelly;Minna, John D.;Bunn, Paul A.
通讯作者：
Bunn, Paul A.
影响因子：
21.3
作者：
Eisner V;Picard M;Hajnóczky G
通讯作者：
Hajnóczky G
DOI：
10.1261/rna.047126.114
发表时间：
2014-12
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Lasda E;Parker R
通讯作者：
Parker R