Abstract：
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 morphological structure and damage of the brain tissue. The immunofluorescence assay was employed to measure the expression of glial fibrillary acidic protein(GFAP) and glial scar. Western blot was employed to determine the protein levels of GFAP, neurocan, phosphacan, 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 impairment, 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, improved 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 inflammatory response, and reducing the formation of glial scar.
摘要：
本研究旨在探讨脑泰方(NTF)对大鼠脑缺血再灌注损伤(CIRI)模型小胶质细胞极化及胶质瘢痕相关蛋白的影响。通过大脑中动脉闭塞/再灌注建立CIRI模型。48只造模成功的大鼠随机分为7d,模型14d，NTF7d,和NTF14d组(n=12)。此外,选择12只SD年夜鼠作为假手术组。NTF组给予27g·kg~(-1)·d~(-1)的NTF悬液,和假，模型7d，模型14d组每天灌胃相同体积的生理盐水，连续7天和14天，分别。干预之后,对隆加评分进行评价。通过2,3,5-三苯基-2H-氯化四唑(TTC)染色测量梗死体积。进行了Morris水迷宫和野外测试，以评估空间学习，记忆,认知功能,和老鼠的焦虑程度。采用苏木精-伊红（HE）染色观察脑组织的形态结构和损伤。采用免疫荧光法检测胶质纤维酸性蛋白（GFAP）和胶质瘢痕的表达。蛋白质印迹用于确定GFAP的蛋白质水平，Neurocan,磷酸盐,CD206,精氨酸酶-1(Arg-1),白细胞介素（IL）-1β，IL-6和IL-4。与假相比，模型7d和模型14d组出现不同程度的脑梗死,大脑皮层和海马的严重病理损伤，神经损伤，减少空间学习和记忆，认知功能障碍，严重的焦虑,星形胶质细胞增生,增厚的半影胶质瘢痕，和上调IL-1β的蛋白水平，IL-6,GFAP,Neurocan,磷酸盐,CD206和Arg-1(P<0.01)。与模型组相比，NTF7d和NTF14d组改善了空间学习，记忆,和认知功能，减少焦虑，改善神经功能，脑梗塞体积减少，减少星形胶质细胞增生，半影神经胶质疤痕变薄，下调GFAP的蛋白质水平，Neurocan,磷酸盐,IL-6和IL-1β，并上调IL-4，CD206和Arg-1的蛋白水平（P＆lt;0.05或P＆lt;0.01）。NTF通过诱导小胶质细胞的M2极化对CIRI发挥神经保护作用，抑制炎症反应，减少胶质瘢痕的形成.
