巴曲酶在小鼠实验性自身免疫性脑脊髓炎中的作用  

Effects of batroxobin on acute experimental autoimmune encephalolmyelitis

作　　者：田书娟 黄德晖[2] 杨扬[2] 王晓[1] 吴卫平[2] TIAN Shujuan;HUANG Dehui;YANG Yang;WANG Xiao;WU Weiping(不详;Department of Neurology,the General Hospital of Chinese PLA,Beijing 100850,China)

机构地区：[1]河北医科大学第一医院神经内科,050031 [2]中国人民解放军总医院神经内科,100850

出　　处：《中国神经免疫学和神经病学杂志》2020年第6期434-441,465,共9页Chinese Journal of Neuroimmunology and Neurology

基　　金：中国人民解放军总医院院内课题(2014BJ-JSCX-2005)。

摘　　要：目的研究巴曲酶对MOG35-55诱导的实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠的影响,探讨纤维蛋白原在神经系统脱髓鞘疾病中的作用机制。方法55只C57BL/6雌性小鼠分为正常对照组、EAE模型组、巴曲酶预防组(预防组)和巴曲酶治疗组(治疗组)4组,后三组使用MOG35-55(200μg,/只腹腔注射)免疫诱发EAE动物模型,预防组于免疫前3 d,治疗组于免疫后3 d连续给予巴曲酶〔30 BU/(kg·只)〕,然后改为隔日给药至发病高峰期(未发病者至免疫后50 d),免疫后每日观察小鼠发病情况并进行神经功能评分。通过常规苏木素-伊红(HE)染色、Luxol fast blue染色、快速银染法分别观察各组小鼠病灶内炎性浸润、髓鞘脱失及轴索损害情况,采用免疫组化方法观察各组小鼠T细胞浸润、纤维蛋白沉积、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、髓鞘碱性蛋白(myelin basic protein,MBP)及基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)表达情况,并应用实时荧光定量PCR技术检测MBP mRNA的表达,RT-PCR技术检测MMP-9 mRNA的表达。结果与EAE模型组比较,预防组和治疗组发病及达峰时间均延迟、神经功能评分降低(均P<0.05),预防组与治疗组比较差异无统计学意义(P>0.05)。与EAE模型组比较,治疗组和预防组小鼠炎性细胞浸润明显减少(均P<0.05),髓鞘脱失及轴索损害轻微;免疫组化检测显示治疗组和预防组小鼠脊髓中T细胞浸润、纤维蛋白沉积及GFAP表达均减少(均P<0.05),MBP和MMP-9表达增多(均P<0.05)。与EAE模型组比较,预防组及治疗组MBP mRNA表达明显上调(P<0.05)。EAE各组MMP-9 mRNA表达较正常对照组上调(均P<0.05),而EAE三组间MMP-9的mRNA表达差异无统计学意义(P>0.05)。结论巴曲酶可能通过减少纤维蛋白沉积、影响胶质细胞活性等多个方面改善EAE小鼠的病理和临床症状。Objective To explore the effect of batroxobin on experimental autoimmune encephalomyelitis(EAE)mice model induced by MOG35-55,and the mechanism of fibrinogen in demyelinating diseases of the nervous system.Methods 55 C57BL/6 female mice were randomly divided into a normal control group,an EAE control group,a batroxobin prevention group,and a batroxobin treatment group.Mice in the latter three groups were immunized with MOG35-55(200μg,intraperitoneal injection)to induce EAE mice model.Batroxobin(30 Bu/kg)was separately injected in 3 days before or after immunization,and then alternate-day dosing until the peak of the disease or 50 days after immunization(undiseased).We observed the incidence and clinical symptom scores.The inflammatory infiltration,myelin loss and axonal damage were observed by hematoxylin-eosin(HE)staining,Luxol fast blue staining and rapid silver staining.Immunohistochemistry was used to observer T cell infiltration,fibrin deposition and the expression levels of glial fibrillary acidic protein(GFAP),myelin basic protein(MBP),and matrix metalloproteinases 9(MMP-9).Real-time PCR was used to detect the MBP mRNA expression and Reverse Transcription-PCR was used to detect the MMP-9 mRNA expression.Results The disease onset and peak time were delayed,the neurological function score was significantly decreased in the batroxobin prevention group and the treatment group as compared with those in the EAE control group(P<0.05),there was no significantly difference between the prevention group and the treatment group(P>0.05).Inflammatory cell infiltration,myelin loss and axonal injury were significantly reduced in the treatment group and the prevention group as compared with those in the EAE control group(P<0.05).Compared with the EAE control group,T cell infiltration,fibrin deposition and GFAP expression in the treatment group and the prevention group were significantly decreased(P<0.05),while the expressions of MBP and MMP-9 were significantly increased(P<0.05).The mRNA expression of MBP in the preventio

关 键 词：脑脊髓炎 自身免疫性 实验性 多发性硬化 巴曲酶 纤维蛋白 基质金属蛋白酶 

分 类 号：R744.51[医药卫生—神经病学与精神病学]