The CD40/CD40L system regulates rat cerebral microvasculature after focal ischemia/reperfusion via the mTOR/S6K signaling pathway
The CD40/CD40L system regulates rat cerebral microvasculature after focal ischemia/reperfusion via the mTOR/S6K signaling pathway
复制标题
CD40/CD40L系统通过mTOR/S6K信号通路调节局灶性缺血/再灌注后大鼠脑微血管
DOI：
10.1080/01616412.2018.1473075
复制
发表时间：
2018-01-01
影响因子：
1.9
通讯作者：
Shi, Hai-Bin
中科院分区：
文献类型：
--
作者：
Jiang, Run-Hao;Xu, Xiao-Quan;Shi, Hai-Bin
摘要目的：CD 40/CD 40配体（CD 40 L）在微血管血栓形成中的作用已被广泛接受。然而，CD 40/CD 40 L系统和可溶性CD 40 L（sCD 40 L）与微血管血栓形成之间的确切机制目前仍是一个深入研究的课题。本研究旨在探讨CD 40/CD 40 L系统调控大鼠局灶性脑缺血/再灌注（I/R）后微血管血栓形成的可能机制。方法：采用大鼠大脑中动脉阻塞（MCAO）模型。实验分为假手术组、MCAO组和MCAO + CD 40拮抗剂组。采用酶联免疫吸附试验（ELISA）检测试剂盒，观察局灶性I/R后0、1、3、5、6、12 h血清sCD 40 L水平的动态变化，并采用流式细胞术和免疫荧光法分别检测血小板表面和血管内皮细胞表面CD 40的表达水平。再灌注12 h后分析脑梗死体积。通过Western印迹测定mTOR/S6 K信号传导。结果如下：MCAO和CD 40拮抗剂治疗大鼠之间血栓形成的比较揭示了CD 40和/或CD 40 L在血小板和血管内皮细胞中炎症增强的血栓形成反应中的作用。MCAO大鼠产生了血栓形成的加速，伴随着血清中CD 40水平的增加。与MCAO模型组相比，CD 40拮抗剂治疗组脑梗死灶明显减少。MACO模型组mTOR/S6 K信号通路较CD 40拮抗剂治疗组激活。结论：本研究结果提示，CD 40/CD 40 L系统参与了MCAO及再灌注后微血管血栓形成和脑梗死的发生。mTOR/S6 K信号通路参与CD 40/CD 40 L对局灶性I/R后脑微血管的调节。缩略语：AKT：蛋白激酶B; CD40L：CD 40配体;脑脊液：脑脊液; FITC：异硫氰酸荧光素;进口/进口：缺血/再灌注; MCAO：大脑中动脉闭塞; mTOR：雷帕霉素的机制靶点;体育部：P-藻红蛋白; sCD 40 L：可溶性形式的CD 40 L; TNF-α：肿瘤坏死因子-α;重量：野生型
ABSTRACT Objective: The role of CD40/CD40 ligand (CD40L) in microvascular thrombosis is now widely accepted. However, the exact mechanisms linking the CD40/CD40L system and the soluble form of CD40L (sCD40L) with microvascular thrombosis are currently a topic of intensive research. The objective of this study was to assess the potential mechanisms in CD40/CD40L system-regulated microvascular thrombosis after focal ischemia/reperfusion (I/R). Methods: Rats were subjected to 60-min transient middle cerebral artery occlusion (MCAO). The experiments were divided into three groups: sham operation, MCAO, and MCAO + CD40 antagonist. Dynamic changes of serum-free sCD40L levels for 0, 1, 3, 5, 6, and 12 h by ELISA detecting kit after focal I/R were observed, and the CD40 expression levels in both platelet surface and vascular endothelial cell surface were measured by flow cytometry and immunofluorescence, respectively. Cerebral infarct volume was analyzed 12 h after reperfusion. mTOR/S6K signaling was determined by Western blot. Results: A comparison of thrombus formation between MCAO and CD40 antagonist treatment rats revealed a role for CD40 and/or CD40L in the inflammation-enhanced thrombosis responses in both of the platelet and vascular endothelial cell. MCAO rats yielded an acceleration of thrombus formation that was accompanied by increased CD40 levels in serum. The brain infarction was significantly decreased in CD40 antagonist treatment group compared to MCAO model group. The mTOR/S6K signaling was activated in MACO model than that of CD40 antagonist treatment group. Conclusions: Our findings indicate that CD40/CD40L system contributes to microvascular thrombosis and brain infarction induced by MCAO and reperfusion. The mTOR/S6K signaling pathway is involved in the regulation of cerebral microvasculature after focal I/R by CD40/CD40L. Abbreviations: AKT: protein kinase B; CD40L: CD40 ligand; CSF: cerebrospinal fluid; FITC: fluorescein isothiocyanate; I/R: ischemia/reperfusion; MCAO: middle cerebral artery occlusion; mTOR: mechanistic target of rapamycin; PE: P-phycoerythrin; sCD40L: soluble form of CD40L; TNF-a: tumor necrosis factor-alpha; WT: wild type.