摘要
目的研究柔肝通络汤对脑缺血再灌注大鼠海马区Notch信号通路不同时间点相关蛋白表达的影响,探讨柔肝通络汤治疗缺血性脑卒中分子调节机制。方法雄性健康SD大鼠164只,根据体重随机分为空白对照组、假手术组、模型组、柔肝通络汤组(柔肝组)。除空白对照组外,其他组采用线栓法制备大鼠局灶型缺血再灌注模型(MCAO)(假手术组不形成梗阻灶)。除柔肝组用柔肝通络汤外余三组均蒸馏水灌胃(1.4g/100g)。每组根据缺血2h再灌注后5个时间点6h、1d、3d、7d、14d按每组6只大鼠分为5个亚组。按照5个时间点进行神经功能缺损评分,取脑组织行TTC染色评估缺血脑损伤面积,予免疫组化(SP法)检测不同时间点Notch1、Hes5、Mash1蛋白表达情况。结果与模型组对比,柔肝组TTC染色脑梗死面积明显低于模型组(P<0.05);与空白组和假手术组对比,术后3~14d模型组、柔肝组Notch1、Hes5、Mash1蛋白表达均上升(P<0.05);与模型组对比:术后3~14d柔肝组神经缺损评分明显下降(P<0.05),术后3~14d柔肝组Notch1、Hes5蛋白表达较模型组增加(P<0.05),两蛋白表达均于术后7d达最高值后逐渐下降;术后3~14d柔肝组Mash1蛋白表达较模型组减少(P<0.05),Mash1蛋白在3d表达增加,表达在7d显著下降,表达于第14天达最高值。结论柔肝通络汤可以通过激活Notch信号通路改善缺血后脑梗死面积和神经功能障碍,在脑缺血损伤早期上调Notch1、Hes5的表达,下调Mash1的表达,干预神经干细胞的细胞周期,发挥神经修复、细胞再生作用。
Objective To study the effect of Rougan Tongluo Decoction on the expression of related proteins at different time points in the Notch signaling pathway in the hippocampus of rats with cerebral ischemia reperfusion,to explore the molecular regu-lation mechanism of Rougan Tongluo Decoction in the treatment of ischemic stroke.Methods 164 healthy male SD rats,Except for the blank Control group,other groups were Subjected to the preparation of focal ischemia-reperfusion(MCAO)using the thread embolism method(the Sham operation group did not form an infarction area).According to weight,they were randomly divided in-to blank control group,sham operation group,model group,Rougan Tongluo decoction group(Rougan group),except for the Rougan group,use Rougan Tongluo Decoction and the remaining three groups were all intragastrically administered with distiled water(1.4g/100g).Each group was divided into 5 subgroups according to the 5 time points 6h,1d,3d,7d,14d after 2h ische-mia and reperfusion,and 6 rats in each group.The neurological deficit was scored at 5 time points,and the brain was taken for TTC staining to assess the area of ischemic brain damage,and immunohistochemistry(SP method)was used to detect the protein expression of Notchl,Hes5,and Mashl at different time points.Results Compared with the model group,the TTC stained cerebral infarction area in the soft liver group was significantly lower than that in the model group(P<0.05).Compared with the blank group and the sham operation group,the protein expressions of Notchl,Hes5,and Mashl in the model group and the soft liver group increased from 3d to 14d after the operation(P<0.05).Compared with the model:3d-14d after operation,the nerve defect score of the soft liver group was significantly decreased(P<0.05),the protein expression of Notchl and Hes5 in the liver softening group increased from 3d to 14d after operation compared with the model group(P<0.05),and both protein expressions decreased gradually after reaching the highest value on the 7th day after operation.The expression of Mashl protein in the soft liver group was lower than that in the model group after 3d-14d(P<0.05),the expression of Mashl protein increased on the 3rd day,and decreased significantly on the 7th day,and reached the highest value on the 14th day.Conclusion Rougan Tongluo Decoction can improve the area of cerebral infarction and neurological dysfunction after ischemia by activating the Notch signaling pathway.Up-regulate the expression of Notchl and Hes and down-regulate the expression of Mashl in the early stage of cerebral ischemia injury,intervene in the cell cycle of neural stem cells,play a role in nerve repair and cell regeneration.
作者
龙华君
张珊珊
李映辰
胡国恒
柏正平
伍大华
LONG Hua-jun;ZHANG Shan-shan;LI Ying-cheng;HU Cuo-heng;BAI Zheng-ping;WU Da-hua(Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Provincial Hospital of Integrated Traditional Chinese and Western Medicine/Afiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 410006,China;First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2024年第8期1793-1798,共6页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81904173)
湖南省卫生健康委科研计划课题(C202303078160)
湖南省长沙市自然科学基金(kq2208145)
2022年全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)。
