高迁移率族蛋白B1介导大鼠创伤性脑损伤后小胶质细胞M1极化和神经炎症  

High mobility group box 1 mediates M1 polarization of microglia and neuroinflammation after traumatic brain injury in rats

作　　者：林维斌 陈祥荣[1] 曾以勒[1] 徐朝阳[1] 陈峻严[1] Lin Weibin;Chen Xiangrong;Zeng Yile;Xu Chaoyang;Chen Junyan(Department of Neurosurgery,the Second Affiliated Hospital,Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院神经外科,泉州362000

出　　处：《中国组织化学与细胞化学杂志》2021年第5期405-411,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：福建省自然科学基金(2019J01168)。

摘　　要：目的观察高迁移率族蛋白B1(high mobility group box 1,HMGB1)通路调控小胶质细胞极化对创伤性脑损伤(traumatic brain injury,TBI)后神经炎症反应的影响,探讨伤后神经炎症可能的干预靶点。方法健康雄性SD大鼠96只,随机分为4组:假手术组(Sham)、脑损伤组(TBI)、脑损伤+溶剂组(TBI+Vehicle)、脑损伤+甘草酸(glycyrrhizic acid,GL)(HMGB1拮抗剂)(TBI+GL)组,再按1、3、7、14 d 4个时间点分设4个亚组,每亚组均为6只。采用改良Feeney自由落体法建立大鼠创伤性脑损伤模型。采用改良神经功能评分(mNSS)对各组大鼠伤后各时间点神经功能进行评价;伤后第3 d Nissl染色检测细胞凋亡率;免疫荧光染色检测挫伤脑组织HMGB1与Iba-1的共表达;酶联免疫吸附测定法(ELISA)检测血清中TNF-α、IL-1β、IL-6、IFN-γ水平;Western blot检测挫伤脑组织HMGB1、CD16、CD206的表达。结果与Sham组比较,脑损伤后大鼠神经元凋亡率上升,神经功能降低,大脑皮层小胶质细胞上Iba-1染色阳性HMGB1蛋白共表达阳性率增加,CD16染色阳性的M1型小胶质细胞显著增多,相关炎症因子TNF-α、IL-1β、IL-6和IFN-γ血清水平也显著增高,而应用GL可明显抑制上述变化。结论在TBI后HMGB1通路参与调控小胶质细胞极化及介导神经炎症过程,抑制HMGB1可促使大鼠TBI后小胶质细胞向M2亚型极化,减轻TBI后神经炎症反应,进而改善伤后神经功能。Objective To observe the effect of high mobility group box 1(HMGB1)pathway on neuroinflammation after traumatic brain injury(TBI)through regulating microglia polarization in the rats,and to explore possible interventional targets for neuroinflammation after TBI.Methods Ninety-six male Sprague-Dawley rats were randomly divided into 4 groups:sham operation(Sham)group,TBI group,TBI+Vehicle group and TBI+glycyrrhizic acid(GL)(HMGB1 antagonist)(TBI+GL)group;four sub-groups(n=6 for each sub-group)were set up according to the time points of day 1,3,7 and 14 for each group.TBI animal models were established by using Feeney’s weight-drop method.Behavioral outcomes of the rats were measured by using modified neurologic severity score(mNSS)system at day 1,3,7 and 14 after TBI to determine neurological function.At day 3 after TBI,apoptosis neurons were detected with Nissl staining;the co-expressions of HMGB1 and Iba-1 in TBI cerebral cortex were detected by immunofluorescence staining;the serum levels of TNF-α,IL-1β,IL-6,IFN-γ,HMGB1 were tested by enzyme linked immunosorbent Assay(ELISA);the expressions of HMGB1,CD16,CD206 in TBI cerebral cortex were determined by Western blotting.Results Compared with the Sham group,the apoptosis rate of neurons was increased,the neurological function was decreased in TBI group,the co-expression positive rate of Iba1 and HMGB1 in the cerebral cortical microglia was increased,the expression of M1 microglia cells(positive for CD16 staining)and the serum levels of TNF-α,IL-1β,IL-6 and IFN-γwere increased significantly after TBI.These changes were effectively inhibited by the application of GL.Conclusion HMGB1 is involved in the process of regulating microglia polarization and neuroinflammation after TBI.Inhibition of HMGB1 can promote the polarization of microglia towards the M2 subtype after TBI in rats and reduce neuroinflammatory response,thus improve neurological outcomes of rats.

关 键 词：创伤性脑损伤 高迁移率族蛋白B1 小胶质细胞 炎症反应 神经保护 

分 类 号：R651.15[医药卫生—外科学]