复方景川片对大鼠缺血性脑损伤的保护作用及对小胶质细胞/巨噬细胞极化的影响

The protective effect of compound Jingchuan tablets on ischemic brain injury and its effect on microglia/macrophage polarization in rats

目的观察复方景川片对大鼠缺血性脑损伤的神经保护作用,并探讨其对小胶质细胞/巨噬细胞极化的影响。方法60只大鼠按照随机数字表法分为空白对照组,模型组,尼莫地平组(阳性对照)和复方景川片低、中、高剂量组。除空白对照组外,其他各组采用线栓法构建大脑中动脉闭塞模型,复方景川片低、中、高剂量组于建模前7 d按照复方景川片0.78、1.56、3.12 g/kg连续灌胃给药10 d。建模后48 h采用Longa神经功能评分和横木行走实验评价大鼠神经行为改变,苏木素-伊红染色观察脑组织病理变化,免疫组化染色及Western blot检测大脑组织中细胞分化簇86(CD86)、精氨酸酶1(Arg1)表达。结果与空白对照组比较,模型组大鼠神经功能评分、横木行走评分显著增高(P<0.05),缺血侧大脑皮质神经元明显坏死、缺失,小胶质细胞/巨噬细胞增生较明显,CD86、Arg1表达均明显增加(P<0.05)。与模型组比较,各给药组大鼠神经功能评分和横木行走实验评分明显降低(P<0.05),缺血侧大脑皮质病变减轻,神经元坏死数目减少,锥体细胞增多,仍可见小胶质细胞/巨噬细胞增生,大脑皮质及脑组织CD86表达降低,Arg1表达增加,尤以高剂量变化更为明显(P<0.05)。结论复方景川片能够减轻大鼠脑缺血后的神经损伤,且高剂量效果更为明显,其神经保护机制可能与其调控小胶质细胞/巨噬细胞极化状态有关。

Objective To observe the protective effect of compound Jingchuan tablets on cerebral ischemic injury in rats and to explore its effect on microglia/macrophage polarization.Methods Sixty rats were randomly divided into sham group,model group,nimodipine group(positive control group),low,middle and high-dose compound Jingchuan tables groups.Except for the sham group,the middle cerebral artery occlusion model rats were established with thread blocking method in the other groups.Seven days before modeling,the low,middle and high-dose groups(0.78,1.56 and 3.12 g/kg of compound Jingchuan tables)were gived continuous intragastric administration for 10 days.Longa neurological function score and cross-walking test score were used to evaluate neurobehavioral changes 48 h after modeling.Hematoxylin-eosin staining was used to observe the pathological changes of brain tissue.Immunohistochemical staining and Western blot assay were used to observe the expression changes of clusters of differentiation 86(CD86)and arginase-1(Arg1).Results Compared with the sham group,the neurological function score and the cross-walk test score were significantly increased in the model group(P<0.05).The neurons of cerebral cortex on the ischemic side were significantly necrotic and missing,and the proliferation of microglia/macrophages was obvious.The expressions of CD86 and Arg1 increased significantly(P<0.05).Compared with the model group,the scores of neurological function and cross-walking test were significantly reduced in each treated group(P<0.05).On the ischemic side,the lesions of cerebral cortex were alleviated,the number of necrotic nerve cells decreased,the number of pyramidal cells increased,and microglia/macrophage were still proliferated.The expression of CD86 decreased,and the expression of Arg1 increased,especially in the high-dose group(P<0.05).Conclusion Compound Jingchuan tablets can reduce nerve damage after cerebral ischemia and the effect is more obvious at high dose.The protective mechanism may be related to the regulation of microglia/macrophage polarization.