柔肝通络汤对MCAO大鼠内源性干细胞表达的影响  

Effects of Rougan Tongluo Decoction on Expressions of Endogenous stem cells in MCAO Rats

作　　者：及晓梦 龙华君[2] 刘雨[2] 张珊珊 帅文昊 杨颖 伍大华[2] Ji Xiaomeng;Long Huajun;Liu Yu;Zhang Shanshan;Shuai Weihao;Yang Ying;Wu Dahua(Hunan University of Chinese Medicine,Hunan,Changsha 410208,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省中医药研究院附属医院,湖南长沙410006

出　　处：《中国中医急症》2020年第6期1015-1019,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：湖南省自然科学青年基金资助项目(2017JJ3187);湖南省教育厅科学研究项目(19C1411);湖南省中医药研究院院级科研项目(201917)。

摘　　要：目的观察柔肝通络汤对缺血再灌注脑损伤大鼠内源性神经干细胞增殖分化表达的影响。方法采用改良Longa法制作大鼠大脑中动脉堵塞(MCAO型)所致缺血再灌注脑损伤模型;将120只SD雄性大鼠随机分为空白组、假手术组、模型组(蒸馏水灌胃)和柔肝通络汤组(柔肝通络汤灌胃),其中按照缺血2 h再灌注6 h、1 d、3 d、7 d、14 d各分为5个亚组,每组6只。动态观察各时间点大鼠神经功能学评分;腹腔注射5-溴脱氧尿嘧啶核苷(BrdU)标记脑组织内增殖细胞,分别采用BrdU/NeuN神经元核抗原(Neu N)、BrdU/GFAP胶质纤维酸性蛋白(GFAP)标记大脑皮质缺血区域新生神经元和星形胶质细胞,并计算阳性细胞数目。结果与模型组比较,柔肝通络汤组术后3、7、14 d神经功能缺损评分明显降低(P<0.05);柔肝通络汤组BrdU、BrdU/NeuN的阳性细胞数目均明显升高(P<0.05),BrdU阳性细胞数在第7日达到高峰,随后呈下降趋势,BrdU/NeuN在第3~7日增加迅速,第7日后增加缓慢;给药后BrdU/GFAP的阳性细胞数较模型组减少(P<0.05)。结论柔肝通络汤可以促进缺血再灌注脑损伤大鼠内源性神经干细胞增殖,并分化为成熟神经元,同时抑制星形胶质细胞的过度增殖,促进神经再生和修复,改善神经功能。Objective:To observe the effect of Ruogan Tongluo Decoction on the proliferation and differentiation of endogenous neural stem cells in rats with ischemia reperfusion injury.Methods:Ischemia reperfusion injury model induced by middle cerebral artery occlusion(MCAO)in rats was established by modified Longa method.120 SD male rats were randomly divided into the blank group,the sham group,the model group(distilled water gavage)and the rougan tongluo decoction(Ruogan Tongluo Decoction gavage)groups.According to the ischemia 2 h reperfusion for 6 h,1 d,3 d,7 d and 14 d,there were 5 subgroups with 6 rats in each group.The neurofunctional scores of rats at each time point were observed dynamically.Intraperitoneally injected 5-bromodeoxy uridine(BrdU)labeled proliferating cells in brain tissue,and BrdU/NeuN neuronal nuclear antigen(Neu N)and BrdU/GFAP glial fibrillary acidicprotein(GFAP)were used to label newborn neurons and astrocytes in cerebral ischemic regions,and the number of positive cells was calculated.Results:Compared with the model group,the neurological defect scores of the ruogan tongluo decoction group were significantly reduced on postoperative days 3,7 and 14(P<0.01).The number of BrdU and BrdU/NeuN positive cells in the rougan tongluo decoction group was significantly increased(P<0.05).The number of BrdU positive cells reached a peak on the 7th day,and then showed a downward trend.BrdU/NeuN increased rapidly on the 3rd to 7th day,but slowly on the 7th day.The number of positive cells of BrdU/GFAP was lower than that of the model group(P<0.05).Conclusion:Ruogan Tongluo Decoction can promote the proliferation and differentiation of endogenous neural stem cells into mature neurons in rats with ischemia reperfusion injury,inhibit the excessive proliferation of astrocytes,promote nerve regeneration and repair,and improve nerve function.

关 键 词：缺血再灌注脑损伤 内源性神经干细胞 BrdU/NeuN BrdU/GFAP 神经再生 柔肝通络汤 大鼠 

分 类 号：R285.5[医药卫生—中药学]