Poldip2 mediates blood-brain barrier disruption and cerebral edema by inducing AQP4 polarity loss in mouse bacterial meningitis model.
Poldip2 mediates blood-brain barrier disruption and cerebral edema by inducing AQP4 polarity loss in mouse bacterial meningitis model.
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DOI：
10.1111/cns.13446
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发表时间：
2020-12
影响因子：
5.5
通讯作者：
Huang J
中科院分区：
文献类型：
--
作者：
Gao M;Lu W;Shu Y;Yang Z;Sun S;Xu J;Gan S;Zhu S;Qiu G;Zhuo F;Xu S;Wang Y;Chen J;Wu X;Huang J
据报道，特异性高度极化水通道蛋白-4（AQP 4）表达在血脑屏障（BBB）完整性和脑水转运平衡中起关键作用。聚合酶δ相互作用蛋白2（Poldip 2）的上调参与了缺血性卒中后BBB破坏的加重。本研究旨在研究小鼠细菌性脑膜炎（BM）模型中Poldip 2介导的BBB破坏和脑水肿形成是否通过诱导AQP 4极性丧失而发生。经后池注射B组溶血性链球菌诱导小鼠骨髓炎模型。在BM诱导后1小时鼻内施用重组人Poldip 2（rh-Poldip 2）。在BM诱导前48小时通过脑室内（i.c.v）注射给予靶向Poldip 2的小干扰核糖核酸（siRNA）。在BM诱导前0.5小时静脉内给予基质金属蛋白酶（MMPs）的特异性抑制剂UK 383367。进行Western印迹、免疫荧光染色、定量真实的时间PCR、神经行为测试、脑含水量测试、伊文思蓝（EB）渗透性测定、透射电子显微镜（TEM）和明胶酶谱法。结果显示，在小鼠骨髓模型中，Poldip 2表达上调，AQP 4极性丧失。Poldip 2 siRNA和UK 383367均能改善BM模型的神经行为学结果，减轻脑水肿，保持BBB的完整性，并减轻AQP 4极性的丧失。Rh-Poldip 2上调MMPs和胶质细胞酸性蛋白（GFAP）的表达，下调β-肌营养不良蛋白聚糖（β-DG）、闭锁小带-1（ZO-1）、闭合蛋白和闭合蛋白-5的表达;而Poldip 2 siRNA下调MMPs和GFAP的表达，上调β-DG、ZO-1、闭合蛋白和闭合蛋白-5。类似地，UK 383367下调GFAP的表达并上调β-DG、ZO-1、occludin和claudin-5的表达。Poldip 2抑制可通过MMPs/β-DG途径减轻AQP 4极性的丧失，从而部分减轻脑水肿，保护BBB的完整性。在小鼠细菌性脑膜炎模型中由Poldip 2诱导的AQP 4极性丧失的示意图。Poldip 2诱导MMPs的上调和活化，MMPs介导β-DG的裂解。由于β-DG负责AQP 4锚定到星形胶质细胞终足，β-DG的降解导致AQP 4极性丧失。在本机制研究中，Poldip 2 siRNA和UK 383367分别用于抑制Poldip 2表达和MMP活性。
Specific highly polarized aquaporin‐4 (AQP4) expression is reported to play a crucial role in blood‐brain barrier (BBB) integrity and brain water transport balance. The upregulation of polymerase δ‐interacting protein 2 (Poldip2) was involved in aggravating BBB disruption following ischemic stroke. This study aimed to investigate whether Poldip2‐mediated BBB disruption and cerebral edema formation in mouse bacterial meningitis (BM) model occur via induction of AQP4 polarity loss. Mouse BM model was induced by injecting mice with group B hemolytic streptococci via posterior cistern. Recombinant human Poldip2 (rh‐Poldip2) was administered intranasally at 1 hour after BM induction. Small interfering ribonucleic acid (siRNA) targeting Poldip2 was administered by intracerebroventricular (i.c.v) injection at 48 hours before BM induction. A specific inhibitor of matrix metalloproteinases (MMPs), UK383367, was administered intravenously at 0.5 hour before BM induction. Western blotting, immunofluorescence staining, quantitative real‐time PCR, neurobehavioral test, brain water content test, Evans blue (EB) permeability assay, transmission electron microscopy (TEM), and gelatin zymography were carried out. The results showed that Poldip2 was upregulated and AQP4 polarity was lost in mouse BM model. Both Poldip2 siRNA and UK383367 improved neurobehavioral outcomes, alleviated brain edema, preserved the integrity of BBB, and relieved the loss of AQP4 polarity in BM model. Rh‐Poldip2 upregulated the expression of MMPs and glial fibrillary acidic protein (GFAP) and downregulated the expression of β‐dystroglycan (β‐DG), zonula occludens‐1 (ZO‐1), occludin, and claudin‐5; whereas Poldip2 siRNA downregulated the expression of MMPs and GFAP, and upregulated β‐DG, ZO‐1, occludin, and claudin‐5. Similarly, UK383367 downregulated the expression of GFAP and upregulated the expression of β‐DG, ZO‐1, occludin, and claudin‐5. Poldip2 inhibition alleviated brain edema and preserved the integrity of BBB partially by relieving the loss of AQP4 polarity via MMPs/β‐DG pathway. Schematic illustration of AQP4 polarity loss being induced by Poldip2 in mouse bacterial meningitis model. Poldip2 induced the upregulation and activation of MMPs, which mediates the cleavage of β‐DG. Due to β‐DG being responsible for AQP4 anchoring to astrocytic endfeet, the degradation of β‐DG results in AQP4 polarity loss. In the present mechanism study, Poldip2 siRNA and the UK383367 were used to inhibit Poldip2 expression and MMP activity, respectively.
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DOI：
10.1186/cc11670
发表时间：
2012-10-04
期刊：
Critical care (London, England)
影响因子：
--
作者：
Cao C;Yu X;Liao Z;Zhu N;Huo H;Wang M;Ji G;She H;Luo Z;Yue S
通讯作者：
Yue S
DOI：
10.1016/j.bbrc.2015.04.084
发表时间：
2015-06-26
影响因子：
3.1
作者：
Kim, YoungDoo;Park, Hyejin;Jung, Yong-Keun
通讯作者：
Jung, Yong-Keun
影响因子：
11.8
作者：
Liu, ZP;Wang, ZG;Olson, EN
通讯作者：
Olson, EN
影响因子：
4.4
作者：
Assentoft, Mette;Larsen, Brian Roland;MacAulay, Nanna
通讯作者：
MacAulay, Nanna
影响因子：
2.6
作者：
Chowdhury, Priyanka;Dey, Payel;Ghosh, Utpal
通讯作者：
Ghosh, Utpal