Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, Wannan Medical College, Wuhu 241000, Anhui, China; Department of Neurology, Wannan Medical College First Affiliated Hospital, Yijishan Hospital, Wuhu 241000, Anhui, China.
Department of Neurology, The First Affiliated Hospital of Anhui Medical University, Hefei 230000, Anhui, China.
J Stroke Cerebrovasc Dis. 2022 Jan;31(1):106213. doi: 10.1016/j.jstrokecerebrovasdis.2021.106213. Epub 2021 Nov 24.
Recent studies have reported that receptor-interacting protein kinase 3 (RIPK3)-dependent necroptosis is related to the pathological process of intracerebral hemorrhage (ICH). Some studies support the view that inhibiting necroptosis is a key mechanism preventing inflammation. Inflammation is a crucial factor contributing to neurological injuries and unfavorable outcomes after ICH. The aim of this study was to clarify the association between necroptosis and monocyte chemoattractant protein-1 (MCP-1)-mediated inflammation and identify a new target for the treatment of ICH.
An ICH model was established in C57BL/6 mice by injecting collagenase IV into the right basal ganglia. The RIPK3 inhibitor GSK872 was administered through intraventricular injection. Then, we assessed brain edema and neurobehavioral function. Western blotting was employed to detect changes in RIPK3, phospho-mixed lineage kinase domain-like protein (p-MLKL), MCP-1, phospho-c-Jun N-terminal kinase (p-JNK) and interleukin 6 (IL-6) levels in the brain tissue. The localization of RIPK3 and MCP-1 was observed using immunofluorescence staining. Co-immunoprecipitation was performed to determine the interaction between RIPK3 and MCP-1.
Compared with the sham group, the levels of RIPK3, p-MLKL, MCP-1, p-JNK and IL-6 were increased post-ICH. GSK872 pretreatment significantly reduced RIPK3, p-MLKL, MCP-1, p-JNK and IL-6 expression, accompanied by mitigated cerebral edema and neurobehavioral defects. RIPK3 and MCP-1 colocalized in the perinuclear region after ICH. We detected the formation of the RIPK3-MCP-1 complex in ICH brain tissue.
There exerted an association between RIPK3 and MCP-1. The inhibition of RIPK3 alleviated MCP-1-mediated inflammation following ICH.
最近的研究报告称，受体相互作用蛋白激酶 3（RIPK3）依赖性坏死性凋亡与脑出血（ICH）的病理过程有关。一些研究支持这样的观点，即抑制坏死性凋亡是预防炎症的关键机制。炎症是导致 ICH 后神经损伤和不良结局的关键因素。本研究旨在阐明坏死性凋亡与单核细胞趋化蛋白-1（MCP-1）介导的炎症之间的关系，并确定治疗 ICH 的新靶点。
通过向右侧基底神经节注射胶原酶 IV 建立 C57BL/6 小鼠 ICH 模型。通过脑室注射给予 RIPK3 抑制剂 GSK872。然后，我们评估了脑水肿和神经行为功能。采用 Western blot 检测脑组织中 RIPK3、磷酸化混合谱系激酶结构域样蛋白（p-MLKL）、MCP-1、磷酸化 c-Jun N 末端激酶（p-JNK）和白细胞介素 6（IL-6）水平的变化。采用免疫荧光染色观察 RIPK3 和 MCP-1 的定位。采用免疫共沉淀检测 RIPK3 和 MCP-1 之间的相互作用。
与假手术组相比，ICH 后 RIPK3、p-MLKL、MCP-1、p-JNK 和 IL-6 水平升高。GSK872 预处理可显著降低 RIPK3、p-MLKL、MCP-1、p-JNK 和 IL-6 的表达，同时减轻脑水肿和神经行为缺陷。ICH 后 RIPK3 和 MCP-1 共定位于核周区。我们在 ICH 脑组织中检测到 RIPK3-MCP-1 复合物的形成。
RIPK3 与 MCP-1 之间存在关联。抑制 RIPK3 可减轻 ICH 后 MCP-1 介导的炎症。
