摘要
目的观察小鼠脑缺血/再灌注后小胶质细胞的形态及其表达的促炎因子及酶类变化,探讨小胶质细胞在小鼠脑缺血/再灌注损伤后炎症反应中的作用。方法 48只清洁级雄性昆明小鼠,采用随机方法分为假手术组和模型组,每组24只,再随机将每组分为3个小组,每组8只。制备小鼠右侧大脑中动脉闭塞1h再灌注24h模型,采用Zea Longa评分法行神经功能缺损评分,用2,3,5-三苯基氯化四氮唑(TTC)染色观察缺血后脑梗死体积;采用免疫组化技术观察脑组织中小胶质细胞形态、数量以及iNOS的表达。用酶联免疫吸附法检测脑组织TNF-α的含量。结果小鼠脑缺血1h再灌注24h可引起脑梗死和神经功能缺损。免疫组化显示假手术组小鼠皮层区小胶质细胞呈分支状,胞体小,阳性细胞数(5.97±1.27)个/HP,模型组小鼠皮层区小胶质细胞分支增多,胞体肥大,阳性细胞数(11.36±1.80)个/HP,两组比较差异有统计学意义(P<0.05)。模型组小鼠皮层区iNOS表达及TNF-α含量高于假手术组,差异有统计学意义(P<0.05)。结论小鼠脑缺血/再灌注可引起小胶质细胞激活,发生形态改变,引起脑内iNOS蛋白表达增加及TNF-α释放增多,从而加重脑缺血/再灌注后神经损伤作用。
Objective To investigate the changes of proinflammatory cytokine and enzymeo of microglia,explore the role of microglia in the ischemia-reperfusion inflammatory injury.Methods Forty eight clean level Kunming mice were randomly divided into two groups：sham group（n=24） and model group（n=24）.And then randomly divided into three subgroups of sham group and model group,each group had 8 mice.A mice model was made by right side of the brain artery occlusion 1 h and reperfusion 24 h model.Zea Longa grading line was used for neural function defect scale,the infarct size was determined by 2,3,5-triphenyiterazoloride（TTC） staining.The morphological change of microglia,the numbers of microglia and iNOS were observed with immunohistochemistry.The enzyme-linked immunosorbent assay（ELlSA） was used to detect the concentration of TNF-α in brain tissue.Results Ischemia 1 h reperfusion 24 h can cause the stable infarction volume and neurological deficit score on the cortex.The cortex of sham-operation group displayed the microglia branches,the small of cyton,and the positive expression cells were（5.97±1.27） ind /HP.The cortex of model group displayed hyperplasia of microglia,the large of the cyton,and the positive expression cells were（11.36±1.80） ind /HP.There were significant differences compared to the sham-operation group（P0.05）.The levels of iNOS and TNF-α in model group were higher than sham group,there was significant difference（P0.05）.Conclusion The cerebral ischemia/reperfusion model can induce microglia activation in the cortex of mouse,cause the increase of the expression of the iNOS protein and release of the TNF-α.Thus aggravating cerebral ischemia reperfusion nerve injury.
出处
《新疆医科大学学报》
CAS
2012年第3期310-314,共5页
Journal of Xinjiang Medical University
基金
国家自然科学基金资助项目(编号:81060104)
