Inhibition of SENP6 restrains cerebral ischemia-reperfusion injury by regulating Annexin-A1 nuclear translocation-associated neuronal apoptosis.
Inhibition of SENP6 restrains cerebral ischemia-reperfusion injury by regulating Annexin-A1 nuclear translocation-associated neuronal apoptosis.
复制标题
抑制SENP6通过调节Annexin-A1核转位相关的神经元凋亡来抑制脑缺血再灌注损伤。
作者：
Xia Q;Mao M;Zeng Z;Luo Z;Zhao Y;Shi J;Li X
原理：先前已提出膜联蛋白-A1（ANXA 1）在缺血性卒中损伤期间的神经元凋亡中起关键作用。我们最近的研究表明，ANXA 1被SUMO化修饰，并且这种修饰在脑缺血后被大大削弱，但其对神经元死亡的影响及其机制尚未完全阐明。研究方法：建立小鼠大脑中动脉闭塞模型和原代培养神经元缺氧缺糖再灌注模型。进行Ni 2 +-NTA琼脂糖亲和下拉测定以确定ANXA 1的SUMO化水平。利用免疫共沉淀分析来探索蛋白质相互作用。通过免疫印迹分析、荧光素酶报告基因检测、实时定量PCR等方法研究其调控机制。采用LDH释放法和TUNEL染色法分别观察神经细胞毒性和细胞凋亡。结果：在这项研究中，我们确定了去SUMO化酶sentrin/SUMO特异性蛋白酶6（SENP 6）作为ANXA 1 SUMO化的负调节因子。值得注意的是，我们发现SENP 6介导的ANXA 1去SUMO化诱导其核转位，并在脑缺血损伤期间触发神经元凋亡。一项机制研究表明，SENP 6介导的ANXA 1去SUMO化促进了ANXA 1的TRPM 7和PKC依赖性磷酸化。此外，阻断SENP 6介导的ANXA 1去SUMO化可抑制p53的转录活性，降低Bid表达，抑制caspase-3通路的激活，减少缺氧缺糖再灌注后原代神经元的凋亡。更重要的是，通过过度表达神经元中的SENP 6催化突变体来抑制SENP 6导致缺血性中风小鼠模型中神经功能的显著改善。结论：总之，本研究的结果鉴定了SENP 6在神经元凋亡中的先前未鉴定的功能，并强烈表明SENP 6抑制可为脑缺血提供治疗益处。
Rationale: Annexin-A1 (ANXA1) has previously been proposed to play a crucial role in neuronal apoptosis during ischemic stroke injury. Our recent study demonstrated that ANXA1 was modified by SUMOylation, and that this modification was greatly weakened after cerebral ischemia, but its effect on neuronal death and the underlying mechanism have not been fully elucidated. Methods: Mice subjected to middle cerebral artery occlusion were established as the animal model and primary cultured neurons treated with oxygen-glucose deprivation and reperfusion was established as the cell model of ischemic stroke. The Ni2+-NTA agarose affinity pull-down assay was carried out to determine the SUMOylation level of ANXA1. Co-immunoprecipitation assays was utilized to explore the protein interaction. Immunoblot analysis, quantitative real-time PCR, Luciferase reporter assay were performed to identify the regulatory mechanism. LDH release and TUNEL staining was performed to investigate the neuronal cytotoxicity and apoptosis, respectively. Results: In this study, we identified the deSUMOylating enzyme sentrin/SUMO-specific protease 6 (SENP6) as a negative regulator of ANXA1 SUMOylation. Notably, we found that SENP6-mediated deSUMOylation of ANXA1 induced its nuclear translocation and triggered neuronal apoptosis during cerebral ischemic injury. A mechanistic study demonstrated that SENP6-mediated deSUMOylation of ANXA1 promoted TRPM7- and PKC-dependent phosphorylation of ANXA1. Furthermore, blocking the deSUMOylation of ANXA1 mediated by SENP6 inhibited the transcriptional activity of p53, decreased Bid expression, suppressed caspase-3 pathway activation and reduced the apoptosis of primary neurons subjected to oxygen-glucose deprivation and reperfusion. More importantly, SENP6 inhibition by overexpression of a SENP6 catalytic mutant in neurons resulted in significant improvement in neurological function in the mouse model of ischemic stroke. Conclusions: Taken together, the results of this study identified a previously unidentified function of SENP6 in neuronal apoptosis and strongly indicated that SENP6 inhibition may provide therapeutic benefits for cerebral ischemia.
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DOI：
10.1177/0271678x17709185
发表时间：
2017-11
期刊：
Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism
影响因子：
--
作者：
Percie du Sert N;Alfieri A;Allan SM;Carswell HV;Deuchar GA;Farr TD;Flecknell P;Gallagher L;Gibson CL;Haley MJ;Macleod MR;McColl BW;McCabe C;Morancho A;Moon LD;O'Neill MJ;Pérez de Puig I;Planas A;Ragan CI;Rosell A;Roy LA;Ryder KO;Simats A;Sena ES;Sutherland BA;Tricklebank MD;Trueman RC;Whitfield L;Wong R;Macrae IM
通讯作者：
Macrae IM
SUMO介导的NLRP3调节调节炎症体活性。
影响因子：
16.6
作者：
Barry R;John SW;Liccardi G;Tenev T;Jaco I;Chen CH;Choi J;Kasperkiewicz P;Fernandes-Alnemri T;Alnemri E;Drag M;Chen Y;Meier P
通讯作者：
Meier P
影响因子：
81.5
作者：
Campbell, Bruce C., V;De Silva, Deidre A.;Donnan, Geoffrey A.
通讯作者：
Donnan, Geoffrey A.
影响因子：
4
作者：
Kunz, Kathrin;Piller, Tanja;Mueller, Stefan
通讯作者：
Mueller, Stefan
影响因子：
16
作者：
Dou H;Huang C;Singh M;Carpenter PB;Yeh ET
通讯作者：
Yeh ET