首页|脑泰方对脑缺血再灌注损伤小胶质细胞极化及胶质瘢痕的影响

脑泰方对脑缺血再灌注损伤小胶质细胞极化及胶质瘢痕的影响

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探讨脑泰方(NTF)对脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)大鼠小胶质细胞极化及胶质瘢痕相关蛋白的影响。用大脑中动脉阻塞/复灌注方法建立大鼠CIRI模型。将造模成功的48只大鼠随机分为脑缺血再灌注模型7 d组(model 7 d)、脑缺血再灌注模型14 d组(model 14 d)、脑泰方+脑缺血再灌注模型7 d组(NTF 7 d)、脑泰方+脑缺血再灌注模型14 d组(NTF 14 d),每组12只,另设12只SD大鼠为假手术组(sham)。NTF治疗组给予NTF混悬液27 g·kg-1·d-1灌胃,sham与model 7 d、model 14 d每天给予同体积生理盐水,连续灌胃7、14 d。干预结束,各组大鼠进行Longa评分;2,3,5-三苯四唑氯(TTC)染色法检测脑梗死面积;水迷宫实验、旷场实验评估大鼠空间学习记忆和认知功能以及焦虑程度;苏木素-伊红(hematoxylin-eosin,HE)染色观察脑组织形态结构及损伤情况;免疫荧光检测活化星形胶质细胞胶质纤维酸性蛋白(glial fi-brillary acidic protein,GFAP)的表达和胶质瘢痕情况;免疫印迹法检测脑皮质瘢痕相关蛋白GFAP、神经蛋白聚糖(neurocan)、磷酸蛋白聚糖(phosphacan),小胶质细胞极化蛋白CD206、精氨酸酶-1(arginase-1,Arg-1)及炎症因子白细胞介素(interleukin,IL)-1β、IL-6、IL-4的表达情况。与sham比较,model 7 d及model 14 d大鼠均出现不同程度的脑梗死,脑皮质及神经元海马组织病理损伤严重,神经功能缺损,空间学习记忆及认知功能障碍及焦虑严重,星形胶质细胞增生及半暗带胶质瘢痕厚度均增加,IL-1(3、IL-6表达升高,脑皮质GFAP、neurocan、phosphacan、CD206、Arg-1蛋白表达升高,差异均有统计学意义(P<0。01)。与模型组比较,NTF 7 d及NTF 14 d大鼠空间学习记忆和认知功能改善,焦虑减轻,神经功能改善,脑梗死体积减少,星形胶质细胞增生及半暗带胶质瘢痕厚度均减少,脑皮质组织GFAP、neurocan、phosphacan、IL-6、IL-1(3表达降低,IL-4、CD206、Arg-1表达增加,差异均有统计学意义(P<0。05或P<0。01)。NTF对CIRI具有神经保护作用,能诱导小胶质细胞向M2型极化,抑制炎症反应,减轻胶质瘢痕的形成。
Effect of Naotaifang on microglial polarization and glial scar following cerebral ischemia reperfusion injury
This study aims to investigate the effect of Naotaifang(NTF)on the proteins associated with microglial polarization and glial scar in the rat model of cerebral ischemia reperfusion injury(CIRI).The CIRI model was established by middle cerebral artery occlusion/reperfusion.The 48 successfully modeled rats were randomized into model 7 d,model 14 d,NTF 7 d,and NTF 14 d groups(n=12).In addition,12 SD rats were selected as the sham group.The NTF group was administrated with NTF suspension at 27 g· kg 1·d-1 by gavage,and the sham,model 7 d,and model 14 d groups were administrated with the same volume of normal saline every day by gavage for 7 and 14 days,respectively.After the intervention,Longa score was evaluated.The infarct volume was measured by 2,3,5-triphenyl-2H-tetrazolium chloride(TTC)staining.Morris water maze and open field tests were carried out to evaluate the spatial learning,memory,cognitive function,and anxiety degree of rats.Hematoxylin-eosin(HE)staining was employed to observe the mor-phological structure and damage of the brain tissue.The immunofluorescence assay was employed to measure the expression of glial fi-brillary acidic protein(GFAP)and glial scar.Western blot was employed to determine the protein levels of GFAP,neurocan,phos-phacan,CD206,arginase-1(Arg-1),interleukin(IL)-1β,IL-6,and IL-4.Compared with the sham,model 7 d and model 14 d groups showed cerebral infarction of different degrees,severe pathological injury of cerebral cortex and hippocampus,neurological im-pairment,reduced spatial learning and memory,cognitive dysfunction,severe anxiety,astrocyte hyperplasia,thickening penumbra glial scar,and up-regulated protein levels of IL-1β,IL-6,GFAP,neurocan,phosphacan,CD206,and Arg-1(P<0.01).Compared with the model group,NTF 7 d and NTF 14 d groups improved spatial learning,memory,and cognitive function,reduced anxiety,im-proved nerve function,reduced cerebral infarction volume,reduced astrocyte hyperplasia,thinned penumbra glial scar,down-regulated the protein levels of GFAP,neurocan,phosphacan,IL-6,and IL-1β,and up-regulated the protein levels of IL-4,CD206,and Arg-1(P<0.05 or P<0.01).NTF exerts a neuroprotective effect on CIRI by inducing the M2 polarization of microglia,inhibiting inflamma-tory response,and reducing the formation of glial scar.

strokecerebral ischemia reperfusionglial scarmicrogliaNaotaifang

刘吉勇、张文丽、廖君、胡立娟、王国佐、葛金文、梅志刚

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湖南中医药大学中医学院中医诊断学湖南省重点实验室,湖南长沙 410208

湖南中医药大学药学院,湖南长沙 410208

湖南中医药大学医学院,湖南长沙 410208

湖南中医药大学第二附属医院,湖南长沙 410005

湖南中医药大学中西医结合学院中西医结合心脑疾病防治湖南省重点实验室,湖南长沙 410208

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中风 脑缺血再灌注 胶质瘢痕 小胶质细胞 脑泰方

国家自然科学基金湖南省自然科学基金面上项目湖南省自然科学基金面上项目湖南省教育厅青年项目湖南省卫生健康委科研项目湖南省大学生创新创业训练计划

821741672021JJ304992023JJ3046422B0382D2023030781702342

2024

中国中药杂志
中国药学会

中国中药杂志

CSTPCD北大核心
影响因子:1.718
ISSN:1001-5302
年,卷(期):2024.49(4)
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