颅脑损伤后HMGB1-RAGE介导血管周细胞脱离血管和血脑屏障损伤的机制研究  

The mechanism of the receptors for advanced glycation end products affecting pericyte detachment from blood vessels after traumatic brain injury

作　　者：杜明皓 李嘉妮 李阳[1] 罗娟 树海峰 DU Minghao;LI Jiani;LI Yang;LUO Juan;SHU Haifeng(School of Medicine,Southwest Jiaotong University,Chengdu 610083;Department of Neurosurgery,Western Theater General Hospital,Chengdu 610083,China)

机构地区：[1]西南交通大学医学院,四川成都610083 [2]西部战区总医院神经外科,四川成都610083

出　　处：《西安交通大学学报（医学版）》2023年第2期221-228,共8页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：四川省科技厅科技创新人才项目(No.2022JDRC0041);西南交大医工结合培育专项(重点项目)(No.2682021ZTPY024)。

摘　　要：目的观察颅脑损伤后皮层血管周细胞的反应性变化并探讨其机制。方法使用可控性皮层撞击动物模型模拟颅脑损伤,通过蛋白免疫印迹检测外伤后不同时间点皮层周细胞标记物表达情况,并通过透射电镜确定颅脑损伤后血管周细胞的生物学行为。通过Western blotting检测外伤后高迁移率族蛋白1(highmobilitygroupbox 1,HMGB1)、晚期糖基化终末产物受体(receptor for advanced glycation end product,RAGE)、核因子κB(nuclear factor kappa B,NF-κB)表达水平,并将实验动物分为FPS-ZM1(一种特异性RAGE受体阻滞剂)注射组和野生型组,利用干湿脑重和透射电镜的方法检测外伤后HMGB1-RAGE对周细胞的影响。培养原代小鼠脑微血管周细胞,添加HMGB1重组蛋白并以同时添加FPS-ZM1的培养周细胞作为对照,离体水平探索HMGB1-RAGE通路对血管周细胞的影响。结果外伤后早期皮层周细胞标记物血小板源性生长因子受体B(platelet-derived growth factor receptor beta,PDGFR-β)、NG2蛋白聚糖(NG2 proteoglycan,NG2)表达水平降低(PDGFR-β,Control vs.CCI 3D P<0.05;NG2,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05),并发现周细胞脱离血管,同时伴有局部血脑屏障开放。外伤后早期皮层HMGB1-RAGE-NF-κB信号通路表达升高(HMGB1,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05;RAGE,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05,Control vs.CCI 3D P<0.05,Control vs.CCI 5D P<0.05,Control vs.CCI 7D P<0.05;NF-κB,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05),阻断RAGE与配体结合后皮层水肿减轻(CCI 6H、CCI 1D均P<0.05)、神经血管单元损伤降低。HMGB1重组蛋白可增加培养周细胞的迁移能力(Control vs.HMGB1 P<0.05,Control vs.HMGB1+FPS-ZM1 P<0.05),且能被FPS-ZM1逆转(HMGB1 vs.HMGB1+FPS-ZM1 P<0.05)。结论颅脑损伤后高水平的HMGB1通过周细胞上RAGE受体介导周细胞脱离血管,并导致局部脑水肿的发生。Objective To observe the reactive change of cortical perivascular cells after craniocerebral injury and explore its mechanism.Methods The controllable cortical impact animal model was used to simulate craniocerebral injury,the expressions of cortical pericyte markers at different time points after trauma were studied by Western blotting,and the biological behavior of vascular pericytes after craniocerebral injury was determined by transmission electron microscopy.Post-traumatic high mobility group box 1(HMGB1),receptor for advanced glycation end product(RAGE),and nuclear factorκB(NF-κB)were detected by Western blotting.The experimental animals were divided into FPS-ZM1(a specific RAGE receptor blocker)injection group and wild-type group.Wet and dry brain weight and transmission electron microscopy were used to study the post-traumatic effects of HMGB1-RAGE on pericytes.The primary mouse brain microvascular pericytes were cultured and supplemented with HMGB1 recombinant protein;the cultured pericytes supplemented with FPS-ZM1 were used as the control to explore the effect of HMGB1-RAGE pathway on vascular pericytes in vitro.Results The expression levels of early posttraumatic cortical pericyte markers platelet-derived growth factor receptor beta(PDGFR-β)and NG2 proteoglycan(NG2)decreased(PDGFR-β,Control vs.CCI 3D P<0.05;NG2,Control vs.CCI 6H P<0.05;Control vs.CCI 1D P<0.05).We found that pericytes were detached from blood vessels,accompanied by local blood-brain barrier opening.The expression of HMGB1-RAGE-NF-κB signaling pathway was increased in the early cortex after trauma(HMGB1,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05;RAGE,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05,Control vs.CCI 3D P<0.05,Control vs.CCI 5D P<0.05,Control vs.CCI 7D P<0.05;NF-κB,Control vs.CCI 6H P<0.05,Control vs.CCI 1D P<0.05).After blocking the binding of RAGE with the ligand,cortical edema was reduced(CCI 6H P<0.05,CCI 1D P<0.05),and neurovascular unit damage was reduced.HMGB1 recombinant protein could increase the mig

关 键 词：血管周细胞 颅脑损伤 晚期糖基化终末产物受体 高迁移率族蛋白1 

分 类 号：R651.1[医药卫生—外科学]