Yue Xuejing, Liu Lixia, Yan Haiqing, Gui Yongkun, Zhao Jun, Zhang Ping
School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453003, Henan, People's Republic of China.
Department of Neurology, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, People's Republic of China.
Neuropsychiatr Dis Treat. 2020 Apr 1;16:891-900. doi: 10.2147/NDT.S228773. eCollection 2020.
IL-12 inhibition of the endothelial cell functions and angiogenesis is mediated by the cross-talk between the lymphocyte and the endothelial cells, which plays a key role in inhibiting the process of angiogenesis in the eyeballs and in malignant tumors.
We established the intracerebral hemorrhage (ICH) rat model, and IL-12 receptor beta monoclonal antibody was injected into the ICH rats. Western blot, immunofluorescence and RT-qPCR were used to detect the gene expression. Brain water content, EB staining, Garcia test, Beam walking test and wire hanging test were used to assess the injury of brain in ICH rats.
IL-12 gene was significantly increase in hematoma border tissue of ICH rats, and IL-12 protein mainly localized in monocytes. Anti-IL-12 treatment with IL-12 monoclonal antibodies could not only significantly decrease the brain water content and EB content in brain tissues of ICH rats, but also significantly increase the score of the Garcia, Beam balance and the Wire hanging test in ICH rats. Moreover, anti-IL-12 treatment significantly decrease the expression of pro-inflammatory gene, inflammatory gene, p-JAK2/JAK2 and p-STAT4/STAT4 protein, but significantly increase the expression anti-inflammatory gene and CD31 protein, and M2 macrophage ratio in hematoma border tissues of ICH rats. In vitro, rmIL-12 inhibited the tube formation of brain microvascular endothelial cells (BMVES) in BMVES and bone marrow-derived monocytes (BMDM) co-culture systems, but not work in a separately cultured BMVES system. In addition, Fedratinib not only reduced p-JAK2/JAK2 and p-STAT4/STAT4 protein expression in BMDM after treating with b-FGF and rmIL-12, but also significantly increased the tube formation of BMVES in BMVES and BMDM co-culture systems after treating with b-FGF and rmIL-12.
Blockade of IL-12 receptor attenuated brain injury after ICH in rat by promoting angiogenesis, and the mechanism might be related to blocking IL-12 could inhibit M2 cell activation via the JAK2/STAT4 pathway.
白细胞介素-12（IL-12）对内皮细胞功能和血管生成的抑制作用是通过淋巴细胞与内皮细胞之间的相互作用介导的，这在抑制眼球和恶性肿瘤的血管生成过程中起关键作用。
我们建立了脑出血（ICH）大鼠模型，并将IL-12受体β单克隆抗体注入ICH大鼠体内。采用蛋白质免疫印迹法、免疫荧光法和逆转录-定量聚合酶链反应（RT-qPCR）检测基因表达。采用脑含水量测定、伊文思蓝（EB）染色、加西亚试验、光束行走试验和悬线试验评估ICH大鼠的脑损伤情况。
ICH大鼠血肿边缘组织中IL-12基因显著增加，IL-12蛋白主要定位于单核细胞。用IL-12单克隆抗体进行抗IL-12治疗不仅可显著降低ICH大鼠脑组织中的脑含水量和EB含量，还可显著提高ICH大鼠的加西亚评分、光束平衡评分和悬线试验评分。此外，抗IL-12治疗可显著降低ICH大鼠血肿边缘组织中促炎基因、炎症基因、磷酸化JAK2（p-JAK2）/JAK2和磷酸化信号转导子和转录激活子4（p-STAT4）/STAT4蛋白的表达，但可显著提高抗炎基因和CD31蛋白的表达以及M2巨噬细胞比例。在体外，重组小鼠IL-12（rmIL-12）在脑微血管内皮细胞（BMVES）和骨髓来源单核细胞（BMDM）共培养体系中抑制BMVES的管腔形成，但在单独培养的BMVES体系中不起作用。此外，fedratinib不仅在经碱性成纤维细胞生长因子（b-FGF）和rmIL-12处理后的BMDM中降低p-JAK2/JAK2和p-STAT4/STAT4蛋白表达，还在经b-FGF和rmIL-12处理后的BMVES和BMDM共培养体系中显著增加BMVES的管腔形成。
阻断IL-12受体可通过促进血管生成减轻大鼠ICH后的脑损伤，其机制可能与阻断IL-12可通过JAK2/STAT4途径抑制M2细胞活化有关。
