重组水蛭素对EAE小鼠脑组织MMP-9表达和NO含量影响的研究  

Effect of hirudin on the expression of MMP-9 and NO in EAE mice

作　　者：罗家明[1] 谭强[1] 陈蓉[2] 蒋国会[1] 王莉[1] 黄敏[1] 刘超[1] 王晓明[1] 秦新月[3] 余巨明[1] 

机构地区：[1]川北医学院附属医院神经内科,四川南充637000 [2]川北医学院微生物学与免疫学教研室,四川南充637000 [3]重庆医科大学附属第一医院神经内科,重庆400016

出　　处：《中风与神经疾病杂志》2015年第7期598-603,共6页Journal of Apoplexy and Nervous Diseases

基　　金：四川省卫生厅资助项目(NO:110305)

摘　　要：目的探讨重组水蛭素(Recombinant hirudin)对实验性自身免疫性脑脊髓炎(EAE)小鼠脑组织基质金属蛋白酶-9(MMP-9)表达和一氧化氮(NO)含量的影响。方法 48只雌性C57BL/6小鼠随机平均分为3组,EAE组运用MOG35-55构建EAE小鼠模型,CFA组由生理盐水代替MOG35-55构建模型,水蛭素组在EAE基础上给予重组水蛭素腹腔注射,CFA组、EAE组给予生理盐水腹腔注射。HE染色和LBF染色观察炎症及脱髓鞘情况,免疫组化检测MMP-9表达,ELLISA检测小鼠脑组织中NO含量,RT-PCR检测MMP-9和诱导型一氧化氮合酶(i NOS)mRNA表达。结果水蛭素组小鼠临床症状、炎症及脱髓鞘程度减轻,与EAE组间的差异有统计学意义(P<0.05)。水蛭素组小鼠脑组织MMP-9蛋白表达和NO含量与EAE组间差异有统计学意义(P<0.05),重组水蛭素组小鼠脑组织iNOS mRNA和MMP-9 mRNA表达与EAE组间差异有统计学意义(P<0.05)。结论重组水蛭素缓解EAE临床症状机制之一可能与抑制EAE小鼠脑组织中MMP-9和iNOS mRNA表达,从而减少MMP-9和NO含量有关。Objective To explore the effect of recombinant hirudin on the expression of Nitric oxide（NO） and（ ma- trix metalloproteinase-9,MMP-9）in EAE mice. Method 48 C57BL/6 mice were divided randomly into three groups e- qually：The EAE group was established by immunizing mice with myelin oligodendrocyte glyeoprotein 35-55 （MOG35-55） , while the CFA group was established by saline instead of MOG35-55. Hirudin group received intraperitoneal injection of re- combinant hirudin. The inflammation status and the demyelination of the mice brain were assessed by Hematoxylin-eosin staining and Luxol Fast Blue（LFB） myelin staining. MMP-9 expression was detected by immunohistochemistry method;and NO was determined by ELISA. The expressions of iNOS mRNA and MMP-9 mRNA in brain tissue were detected by real- time PCR technology. Results Mice treated with hirudin showed dramatic improvement in disease severity （ P 〈 0.05 ）. The inflammation and demyelination of the brain tissue in hirudin group were reduced than that in EAE group（P 〈0.05 ）. The level of NO and expression of MMP-9 in hirudin group were decreased than that in EAE group（ P 〈 0.05 ）, and the ex- pression level of iNOS mRNA and MMP-9 mRNA in hirudin group were decreased than that in EAE group （ P 〈 0.05 ）. Conclusion Recombinant hirudin inhibited the expression level of MMP-9 and NO in EAE mice, which may be parts of therapeutic effects of hirudin.

关 键 词：多发性硬化 实验性自身免疫性脑脊髓炎 重组水蛭素 一氧化氮 基质金属蛋白酶-9 

分 类 号：R744.5[医药卫生—神经病学与精神病学]