大承气汤对脑出血模型大鼠小胶质细胞的作用及机制探讨  

Effect and Mechanisms of Dachengqi Decoction on Microglia of Rats with Intracerebral Hemorrhage

作　　者：杨树升 林丽 Yang Shusheng;Lin Li(Department of Traditional Chinese Medicine,Wuhan Red Cross Hospital Wuhan 430015,China;Laboratory of Medical Molecular and Cellular Biology,College of Basic Medical sciences,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]武汉市红十字会医院中医科,武汉430015 [2]湖北中医药大学基础医学院医学分子细胞生物学实验室,武汉430065

出　　处：《华中科技大学学报（医学版）》2021年第6期728-733,749,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省卫生健康委员会面上项目(No.WJ2019H417);武汉市卫生健康委员会重点项目(No.WZ19A10)。

摘　　要：目的探讨大承气汤对脑出血模型大鼠小胶质细胞及IL-1β、IL-10等炎症因子的影响及分子机制。方法应用胶原酶Ⅶ对成年SD雄性大鼠左侧尾壳核进行脑立体定位注射,建立脑出血大鼠模型。用大承气汤干预1周[立体定位术后6 h开始灌胃,根据前期实验结果使用剂量为1 g/(100 g体重·次),2次/d],采用平衡木实验和悬尾实验评价大鼠脑出血后1、3、7、14和28 d时神经功能缺损情况;采用免疫荧光双重标记观察大承气汤对出血灶周围小胶质细胞和星形胶质细胞形态和数目的影响;ELISA法检测脑出血后14 d出血区IL-1β和IL-10的水平;Western blot法检测脑出血后14 d出血区p38和P-p38水平。结果大承气汤能有效逆转脑出血模型大鼠脑出血后神经功能受损,减少小胶质细胞增生,可逆转IL-1β水平的上升并增加IL-10的表达水平,同时可逆转P-p38蛋白水平的上升。结论大承气汤可通过调节小胶质细胞的数量、形态和功能,改善脑出血后大鼠神经功能缺损情况,其机制可能涉及调控p38 MAPK信号通路而发挥神经保护作用。Objective To investigate the effects and mechanism of Dachengqi decoction on microglia,interleukin-1 beta(IL-1β)and interleukin-10(IL-10)of rats with intracerebral hemorrhage(ICH).Methods To establish the ICH model,the male Sprague-Dawley(SD)rats were injected with collagenaseⅦin the left caudate putamen(CPU).Intervention with Dachengqi decoction for one week(Intragastric administration was carried out 6 hours after stereotactic operation,1 g/100 g body weight,2 times/day).The balance beam test and tail suspension test(TST)were used to assess the neurologic deficits 1,3,7,14 and 28 days after ICH.Double-immunofluorescent labeling was used to analyze the variation of microglia and astrocytes in brain.ELISA was used to detect the levels of IL-1βand IL-10 in the bleeding area on the 14 th day.Western blotting was used to detect the levels of p38 and P-p38 in the bleeding area on the 14 th day.Results Dachengqi decoction could effectively reverse the damage of neural function and reduce the microglia proliferation in ICH rats,down-regulate the level of IL-1βand up-regulate the level of IL-10 in the bleeding area,as well as decrease the level of P-p38.Conclusion Dachengqi decoction plays a neuroprotective role through regulating the number,morphology and function of microglia,which is related to the p38 MAPK pathway.

关 键 词：大承气汤 脑出血 小胶质细胞 白细胞介素 p38 MAPK信号通路 

分 类 号：R743.34[医药卫生—神经病学与精神病学]