摘要
目的探讨丁苯酞(3-n-butylphthalide,NBP)调控小胶质细胞的极化反应在创伤性脑损伤(traumatic brain injury,TBI)中的神经保护作用。方法27只雄性C57BL/6小鼠随机分配(每组9只):假手术组(Sham组)、TBI组和NBP组。Sham组和TBI组给予植物油,NBP组给予等体积100 mg/kg NBP,连续7 d。TBI 3 d后,干湿法评估小鼠脑水肿情况。应用改良版小鼠神经功能缺损评分(modified neurological severity score,mNSS)和转棒疲劳试验检测小鼠损伤后第1、3、7天的神经运动功能恢复情况。免疫组织化学法检测损伤皮层周围区抗离子钙结合适配器分子1(ionized calcium binding adapter molecule 1,Iba1)标记的小胶质细胞活化情况,免疫荧光染色检测胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)标记的星形胶质细胞活化情况。通过精氨酸酶1(arginase-1,Arg1)与Iba1,诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)与Iba1双重染色检测损伤周围区小胶质细胞M1型标志物iNOS及M2型标志物Arg1表达量;髓鞘碱性蛋白(myelinbasic protein,MBP)检测髓鞘丢失情况。结果在mNSS实验中,与TBI组相比,NBP组分别在损伤后的第1、3、7天,小鼠的神经功能评分逐渐降低(P第1天=0.012,P第3天=0.020,P第7天=0.046)。与TBI组相比,匀速转棒实验中,NBP组小鼠停留时间在第3天明显增加(P第3天=0.031);随NBP治疗时间延长,小鼠在第3、7天的匀加速转棒中停留时间逐渐增加(P第3天=0.024,P第7天=0.039)。进一步病理学检测发现,与TBI组相比,NBP组损伤区MBP髓鞘完整性部分恢复(P=0.026);小胶质细胞和星形胶质细胞活化标志物Iba1和GFAP表达降低(PIba1=0.007,PGFAP=0.006)。与TBI组相比,NBP组M2型的小胶质细胞标志物Arg1增加(PArg1/Iba1=0.004),M1型小胶质细胞标志物iNOS减少(PiNOS/Iba1=0.012)。结论NBP可以部分改善TBI小鼠神经运动功能障碍,抑制小胶质细胞和星形胶质细胞的活化,促进小胶质细胞向M2表型的极化。
Objective To investigate the neuroprotective effect of 3-n-butylphthalide(NBP)by regulating microglial polarization in traumatic brain injury(TBI).Methods Twenty-seven male C57BL/6 mice were randomly assigned to Sham group,TBI group and NBP group.Sham group and TBI group were given vegetable oil,and NBP group was given an equal volume of 100 mg/kg butylphthalein for 7 consecutive days.After 3 days,the brain edema of the mice was evaluated by wet and dry method.The modified neurological severity score(mNSS)and fatigue rotarod test were used to detect the recovery of neuromotor function in mice at 1,3 and 7 days after TBI.The activation of ionized calcium binding adapter molecule 1(Iba1)-labeled microglia was verified by immunohistochemistry,and the activation of glial fibrillary acidic protein(GFAP)-labeled astrocytes was confirmed by immunofluorescence staining.Through co-locating with Iba1,the expression of inducible nitric-oxide synthase(iNOS),M1 polarization marker,and arginase-1(Arg1),M2 polarization marker,were indicated by immunofluorescence stainin.Myelinbasic protein(MBP)immunofluorescence staining was used to detect myelin loss.Results In the mNSS experiment,compared with the TBI group,the neural function scores of mice in the NBP group were gradually decreased on day 1,day 3 and day 7 after injury(Pday 1=0.012,Pday 3=0.020,Pday 7=0.046).In constant rotarod test the stick rotation residence time in NBP group was significantly increased on day 3 compared with TBI group(Pday 3=0.031);With the extension of NBP treatment time,the rotation retention time of mice increased in the uniformly accelerated rod rotation experiment on day 3 and day 7(Pday 3=0.024,Pday 7=0.039).Further pathological examination showed that,compared with TBI,the MBP marked-myelin integrity was partially recovered in NBP group(P=0.026).Expressions of microglia and astrocyte activation markers Iba1 and GFAP were decreased(PIba1=0.007,PGFAP=0.006).Compared with TBI group,M2-type microglial marker Arg1 increased(PArg1/Iba1=0.004)and M1-type
作者
郭温晗
李杭
黄成科
许凤燕
方力群
GUO Wenhan;LI Hang;HUANG Chengke;XU Fengyan;FANG Liqun(Department of Human Anatomy,Harbin Medical University,Harbin 150081,China;Department of Neurology,the Fourth Affiliated Hospital of Harbin Medical University,Harbin 150001,China;School of Basic Medicine,Harbin Medical University,Harbin 150081,China)
出处
《国际免疫学杂志》
CAS
2024年第2期136-142,共7页
International Journal of Immunology
基金
哈尔滨医科大学附属第四医院院级特别资助项目(HYDSYKJCXXR)
石药集团恩必普药业有限公司联合项目
中华社会救助基金会项目。
关键词
丁苯酞
创伤性脑损伤
神经炎症
小胶质细胞极化
3-n-butylphthalein
Traumatic brain injury
Neuroinflammation
Microglial polarization
