RIP3 facilitates necroptosis through CaMKII and AIF after intracerebral hemorrhage in mice
RIP3 facilitates necroptosis through CaMKII and AIF after intracerebral hemorrhage in mice
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RIP3通过CaMKII和AIF促进小鼠脑出血后坏死性凋亡
DOI：
10.1016/j.neulet.2021.135699
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发表时间：
2021-02-26
影响因子：
2.5
通讯作者：
Lv，Kun
中科院分区：
文献类型：
--
作者：
Xu，Yang;Wu，Xiaodong;Lv，Kun
研究背景脑出血（intracerebral hemorrhage，ICH）后神经元的坏死性损伤是近年来研究的热点。以往的研究报道，RIP 3及其复合物被认为是坏死性凋亡的中枢介质。在这项研究中，RIP 3在激活CaMKII和AIF的作用investigated.MethodsWe诱导ICH在C57 BL/6小鼠通过注射胶原酶IV到基底神经节。用mPTP抑制剂CsA和CAMKII抑制剂Kn-93、RIP 3 siRNA或RIP 3 rAAV预处理ICH小鼠。评估脑水肿和神经行为。RIP 3、p-MLKL、AIF和CaMKII蛋白的表达通过免疫印迹、免疫荧光（IF）和免疫沉淀（IP）进行评估。结果RIP 3、p-MLKL、CaMKII和AIF在ICH小鼠中的表达显著增加，并且RIP 3-AIF共定位于细胞核。RIP 3的rAAV过表达上调AIF在细胞质和细胞核中的表达，而CaMK II的表达在细胞质中增加。在脑出血损伤后检测RIP 3-AIF和RIP 3-CaMK II的相互作用。这些复合物被抑制CsA与Kn-93或RIP 3 siRNA预处理，从而减少脑水肿和neurological deficits.ConclusionsOur研究结果显示，ICH通过RIP 3-CaMK II复合物和RIP 3-AIF信号通路诱导坏死性神经元死亡。阻断mPTP开放可抑制坏死性凋亡的发生。
BackgroundNecroptosis-induced neuronal damage after intracerebral hemorrhage (ICH) has been documented recently. Previous studies have reported that RIP3 and its complex are recognized as central mediators of necroptosis. In this study, the role of RIP3 in the activation of CaMKII and AIF was investigated.MethodsWe induced ICH in C57BL/6 mice by injecting collagenase IV into the basal ganglia. ICH mice were pretreated with the mPTP inhibitor CsA and the CAMKII inhibitor Kn-93, RIP3 siRNA or RIP3 rAAV. Brain edema and neurobehavior were evaluated. The expression of RIP3, p-MLKL, AIF, and CaMKII proteins was evaluated by western blotting, immunofluorescence (IF) and immunoprecipitation (IP).ResultsSignificant increases in RIP3, p-MLKL, CaMKII and AIF expression were observed in ICH mice, and RIP3-AIF colocalized in the nucleus. Overexpression of RIP3 by rAAV upregulated AIF expression in both the cytoplasm and nucleus, while CaMKII expression was increased in the cytoplasm. The interaction of RIP3-AIF and RIP3-CaMKII was detected after ICH injury. These complexes were inhibited by CsA with Kn-93 or RIP3 siRNA pretreatment, which reduced brain edema and neurological deficits.ConclusionsOur findings revealed that ICH induced necroptotic neuronal death through the RIP3-CaMKII complex and the RIP3-AIF signaling pathway. Moreover, blockade of mPTP opening could suppress the pathogenesis of necroptosis.