摘要
目的观察芪仙通络方不同剂量对大脑中动脉栓塞大鼠髓鞘再生及髓鞘碱性蛋白(MBP)表达的影响。方法大鼠随机分为假手术组、模型组、芪仙通络方高剂量组(31.32 g/kg)、中剂量组(15.66 g/kg)、低剂量组(7.83 g/kg)和丁苯酞组,线栓法制备大脑中动脉阻塞模型,造模后3 d后开始灌胃给药,同时腹腔注射5-乙炔基-2′-脱氧尿苷(EDU)标记脑内增殖细胞,每日1次,连续12 d。于造模后第3、7、14日对大鼠神经功能进行评分;采用罗克沙尔坚牢蓝(LFB)染色观察大鼠髓鞘的病理形态学改变;免疫荧光染色观察大鼠缺血侧EDU/少突胶质细胞转录因子2(Olig2)双标阳性细胞数;免疫组化检测大鼠缺血侧MBP蛋白表达。结果与假手术组比较,模型组大鼠各时间点神经功能评分均明显升高(P<0.01),LFB染色平均光密度值降低(P<0.01),大鼠缺血侧EDU/Olig2双标阳性细胞数相当,差异无统计学意义(P>0.05),MBP蛋白表达显著降低(P<0.01);与模型组比较,各给药组大鼠第7、14 d神经功能评分明显降低(P<0.05),LFB染色平均光密度值明显升高(P<0.05),大鼠缺血侧EDU/Olig2双标阳性细胞数明显增多(P<0.01),MBP蛋白表达显著上调(P<0.01),高剂量组作用最显著(P<0.01)。结论芪仙通络方可改善脑梗死大鼠髓鞘再生功能,其机制可能与促进缺血侧OPCs迁移、分化及MBP蛋白表达有关,且具有浓度依赖性。
Objective:To observe the effects of different doses of Qixian Tongluo Recipe on remyelination and myelin basic protein(MBP)expression in rats with middle cerebral artery occlusion.Methods:Rats were randomly divided into sham-operation group,model group,Qixian Tongluo Recipe 15.66,31.32,7.83 g/kg groups and Butylphthalein group.The suture method was used to replicate the middle cerebral artery occlusion model in rats.The intragastric administration was started on the 3 rd day after modeling,and 5-acetyl-2′-deoxyuridine(EDU)was injected intraperitoneally to label proliferating cells in the brain once a day for 12 consecutive days.The neurological function of rats was evaluated 3 rd,7 th and 14 th days after model establishment;the pathomorphological changes of rat myelin sheath were observed by Luxol Fast Blue(LFB)staining;immunofluorescence staining was used to observed EDU/oliendrocyte transcription factor 2(Olig2)double-labeled positive cells in the ischemic side;immunohistochemistry was used to detect in the ischemic side the expression of MBP.Results:Compared with the sham-operation group,the neurological function score of the model group was prominently increased at all time points(P<0.01),the average optical density of LFB staining decreased(P<0.01),the number of EDU/Oligo2 double-labeled positive cells in the ischemic side of the rats was the similar,without statistical significance(P>0.05);the expression of MBP protein was significantly decreased(P<0.01).Compared with model group,the nerval function score of rats in each administration groups was significantly decreased on day 7 th and 14 th(P<0.05),the average optical density of LFB staining remarkable increased(P<0.05),the number of EDU/Oligo2 double-label positive cells in ischemia side of rats was significantly increased(P<0.01),the expression of MBP protein wassignificantly up-regulated(P<0.01),and the effect of high dose group was the most significan(tP<0.01).Conclusion:Qixian Tongluo Recipe can improve the regeneration of remyelination in rats with cerebral infarction,and themechanism may be related to promoting the migration and differentiation of OPCs on the ischemic side,and the expression of myelin basic protein,also it is concentration dependent.
作者
黄佳
赵建涛
罗晓玲
张心怡
周胜强
刘芳
Huang Jia;Zhao Jiantao;Luo Xiaoling;Zhang Xinyi;Zhou Shengqiang;Liu Fang(Hunan University of Chinese Medicine,Hunan,Changsha 410203,China)
出处
《中国中医急症》
2022年第1期62-66,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
国家中医药管理局“国医大师刘祖贻传承工作室建设项目”
湖南省教育厅重点项目(18A225)
湖南省教育厅优秀青年项目(18B253)
湖南省中医药管理局一般课题(2021083)
湖南省“国内一流培育学科”中西医结合开放基金项目(2019ZXYJH01)
湖南省创新型省份建设专项科普专题(2020ZK4068)
湖南省中医药研究院科研课题(201902)。
关键词
脑梗死
髓鞘再生
髓鞘碱性蛋白
芪仙通络方
大鼠
Cerebral infarction
Remyelination
Myelin basic protein
Qixian Tongluo Recipe
Rats
