小鼠脑出血后不同时间点小胶质细胞极化状态的实验研究  

Experimental Study on the Polarization State of Microglia at Different Time Points after Intracerebral Hemorrhage in Mice

作　　者：武翠梅 王改青[2] 要振佳 WU Cui-Mei;WANG Gai-Qing;YAO Zhen-Jia(The Second Clinical School of Shanxi Medical University,Taiyuan 030000,China;Department of Neurology,The Second Hospital of Shanxi Medical University,Taiyuan 030000,China)

机构地区：[1]山西医科大学第二临床医学院,太原030000 [2]山西医科大学第二医院神经内科,太原030000

出　　处：《中国卒中杂志》2020年第10期1094-1100,共7页Chinese Journal of Stroke

基　　金：国家自然科学基金面上项目(81771294)。

摘　　要：目的观察小鼠脑出血后不同时间点小胶质细胞M1及M2型的转化,为促炎型M1型小胶质细胞向抗炎修复型M2型小胶质细胞的转化,减轻脑出血后神经功能损伤提供理论依据。方法选取健康雄性ICR小鼠48只,随机分为假手术组、脑出血组,每组按术后时间点不同随机分为1 d、3 d、7 d三个时间点,每个时间点8只。通过立体定位仪用微量注射器向尾状核注射Ⅳ型胶原酶0.5 U制备脑出血模型,假手术组注射等量生理盐水。各组于术后对应时间点参照改良Garcia评分量表进行神经功能缺损评分后灌注取脑,采用蛋白免疫印迹检测M1型小胶质细胞标志物肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6),M2型小胶质细胞标志物脑源性神经营养因子(brai n-derived neurotrophic factor,BDNF)、胰岛素样生长因子1(insulinlike growth factor 1,IGF-1)的含量;采用免疫荧光染色标记小胶质细胞M1型(Iba1+CD80)、M2型(Iba1+CD206),评价出血后血肿周围组织小胶质细胞活化状态。结果脑出血组1 d、3 d、7 d各时间点Garci a评分均较假手术组低,TNF-α、IL-6、BDNF及I GF-1的蛋白表达量均较假手术组增多(均P<0.01)。脑出血后1 d时M1型高于M2型小胶质细胞数量(38.33±1.53 vs 23.00±3.00,P=0.01);3 d时M1型同样高于M2型(66.33±3.06 vs 57.33±2.52,P=0.02);7 d时M1型低于M2型(33.67±1.15 vs 52.33±0.58,P<0.01)。结论脑出血急性期(1~3 d)以M1型小胶质细胞为主,脑出血亚急性期(7 d)以M2型小胶质细胞为主。Objective To investigate the transformation of microglia typeⅠand typeⅡat different time points after intracerebral hemorrhage(ICH)in mice,and to provide a theoretical basis for reducing the neurological damage after ICH by transforming pro-inflammatory microglia typeⅠ(M1)into anti-inflammatory repair microglia typeⅡ(M2).Methods Forty-eight healthy male ICR mice were randomly divided into sham operation group and ICH group.Each group was randomly divided into three subgroups according to the different postoperation time points of day 1,day 3,and day 7,with 8 mice in each subgroup.The ICH model was prepared by injectingⅣcollagenase 0.5 U into caudate nucleus through a stereotactic locator,while the mice in sham group were injected with the same amount of normal saline.The neurological deficit of the mice of two groups were assessed using the modified Garcia score at each time point,then the brain was perfused and brain tissue was taken.Western blot was used to detect the contents of microglia M1 markers tumor necrosis factor alpha(TNF-α),interleukin-6(IL-6),and M2 markers brain-derived neurotrophic factor(BDNF),insulin-like growth factor 1(IGF-1);microglia M1(Iba1+CD80)and M2(Iba1+CD206)were labeled by immunofluorescence staining to evaluate the activation state of microglia around the hematoma after hemorrhage.Results At day 1,day 3 and day 7,the Garcia score of the ICH group was lower than that of the sham operation group,and the protein expression of TNF-α,IL-6,BDNF and IGF-1 were higher than that of the sham operation group(all P<0.01).The amount of microglia M1 was higher than M2 at day 1(38.33±1.53 vs 23.00±3.00,P=0.01)and day 3(66.33±3.06 vs 57.33±2.52,P=0.02),while lower than M2 at day 7(33.67±1.15 vs 52.33±0.58,P<0.01).Conclusions Microglia was mainly typeⅠin acute phase of ICH(1-3 day after ICH),and mainly typeⅡin subacute phase(7 day after ICH).

关 键 词：脑出血 小胶质细胞极化 小胶质细胞Ⅰ型 小胶质细胞Ⅱ型 

分 类 号：R743.34[医药卫生—神经病学与精神病学]