Bai Qingqing, Wang Shuoyang, Rao Dongmei, Zhou Zhiming, Wang Jianfei, Wang Qi, Qin Yu, Chu Zhaohu, Zhao Shoucai, Yu Dijing, Xu Yang
Department of Neurology, First Affiliated Hospital of Wannan Medical College, Yijishan Hospital, Wuhu, Anhui, China.
Anhui Province Key Laboratory of Non-coding RNA Basic and Clinical Transformation, Wannan Medical College, Wuhu, Anhui, China.
CNS Neurosci Ther. 2024 Jan;30(1):e14397. doi: 10.1111/cns.14397. Epub 2023 Aug 8.
Necroptosis induced by receptor-interacting protein kinase 3 (RIPK3) is engaged in intracerebral hemorrhage (ICH) pathology. In this study, we explored the impact of RIPK3 activation on neuronal necroptosis and the mechanism of the death domain-associated protein (DAXX)-mediated nuclear necroptosis pathway after ICH.
Potential molecules linked to the progression of ICH were discovered using RNA sequencing. The level of DAXX was assessed by quantitative real-time PCR, ELISA, and western blotting. DAXX localization was determined by immunofluorescence and immunoprecipitation assays. The RIPK3 inhibitor GSK872 and DAXX knockdown with shRNA-DAXX were used to examine the nuclear necroptosis pathway associated with ICH. Neurobehavioral deficit assessments were performed.
DAXX was increased in patients and mice after ICH. In an ICH mouse model, shRNA-DAXX reduced brain water content and alleviated neurologic impairments. GSK872 administration reduced the expression of DAXX. shRNA-DAXX inhibited the expression of p-MLKL. Immunofluorescence and immunoprecipitation assays showed that RIPK3 and AIF translocated into the nucleus and then bound with nuclear DAXX.
RIPK3 revitalization promoted neuronal necroptosis in ICH mice, partially through the DAXX signaling pathway. RIPK3 and AIF interacted with nuclear DAXX to aggravate ICH injury.
受体相互作用蛋白激酶3（RIPK3）诱导的坏死性凋亡参与脑出血（ICH）的病理过程。在本研究中，我们探讨了RIPK3激活对神经元坏死性凋亡的影响以及ICH后死亡结构域相关蛋白（DAXX）介导的核坏死性凋亡途径的机制。
使用RNA测序发现与ICH进展相关的潜在分子。通过定量实时PCR、ELISA和蛋白质印迹法评估DAXX的水平。通过免疫荧光和免疫沉淀试验确定DAXX的定位。使用RIPK3抑制剂GSK872和shRNA-DAXX敲低DAXX来研究与ICH相关的核坏死性凋亡途径。进行神经行为缺陷评估。
ICH后患者和小鼠的DAXX增加。在ICH小鼠模型中，shRNA-DAXX降低了脑含水量并减轻了神经功能障碍。给予GSK872降低了DAXX的表达。shRNA-DAXX抑制了p-MLKL的表达。免疫荧光和免疫沉淀试验表明，RIPK3和AIF易位到细胞核中，然后与核DAXX结合。
RIPK3激活促进ICH小鼠的神经元坏死性凋亡，部分通过DAXX信号通路。RIPK3和AIF与核DAXX相互作用加重ICH损伤。
