摘要
目的:探讨在"神经血管小生境"(neurovascular niche)微环境中,化瘀通络灸对血管性痴呆(vascular dementia,VD)大鼠学习记忆能力及脑源性神经营养因子(brain derived neurotrophic factor,BDNF)、酪氨酸激酶B(tyrosine kinase B,TrkB)表达水平的影响。方法:通过双侧颈总动脉永久性结扎法(2-VO)复制VD大鼠模型,将慢病毒介导的逆转录病毒携带增强绿色荧光蛋白(EGFP)标记的神经干细胞(NSCs)与内皮祖细胞(EPCs)共培养构建"NSCs+EPCs"共植体,移植入VD大鼠侧脑室,建立具有"neurovascular niche"的VD大鼠模型。实验一将造模成功的大鼠分为3组:NSCs+EPCs艾灸组、NSCs+EPCs空白组、模型组,每组12只,模型组和NSCs+EPCs空白组不进行其他任何治疗,NSCs+EPCs艾灸组进行化瘀通络灸法治疗,悬灸"百会""大椎""神庭",每穴20 min,每天治疗1次,14 d为一疗程,共治疗3个疗程。治疗结束后,采用Morris水迷宫实验检测各组大鼠学习记忆成绩;实验二将造模成功的大鼠分为3组:NSCs+EPCs艾灸组、NSCs+EPCs空白组、模型组,每组18只,每一组再根据治疗的不同疗程分为3个亚组,每亚组6只,治疗方法同实验一。每一亚组大鼠再分别于治疗相应疗程后灌注取脑,并行冰冻切片,进行免疫荧光实验观察BDNF/TrkB表达水平。结果:经治疗后,NSCs+EPCs艾灸组在降低逃避潜伏期、减短第一次穿越平台时间、增加穿越平台次数均优于模型组、NSCs+EPCs空白组(P<0.01,P<0.05);NSCs+EPCs艾灸组3个疗程之间蛋白表达存在逐步增加的趋势,疗程之间比较差异有统计学意义(均P<0.05),且NSCs+EPCs艾灸组蛋白表达增加均优于NSCs+EPCs空白组(P<0.05,P<0.01)。结论:化瘀通络灸是临床治疗VD的一种有效方法;化瘀通络灸可上调"neurovascular niche"内BDNF/TrkB蛋白的表达,共同调控"NSCs-EPCs"偶联机制,促进神经新生,修复神经损伤。
Objective To explore the effects of huayu tongluo(resolving stasis, promoting collateral circulation) moxibustion on learning and memory ability and the expressions of brain derived neurotrophic factor(BDNF) and tyrosine kinase B(TrkB) in the rats of vascular dementia(VD) in the microenvironment of neurovascular niche. Methods Using 2-vessel occlusion(2-VO), the VD rat models were duplicated. The neural stem cells(NSCs) labeled with lentiviral vector-mediated enhanced green fluorescent protein(EGFP) were co-cultured with endothelial progenitor cells(EPCs) to structure the NSCs + EPCs implant. The implant was transplanted into the lateral ventricle of VD rats and the VD rat models with neurovascular niche were established. In No.1 experiment, the successful-modeled rats were divided into 3 groups, i.e. a NSCs + EPCs moxibustion group, a NSCs + EPCs blank group and a model group, 12 rats in each one. No any treatment was provided in the model group and the NSCs + EPCs blank group. The huayu tongluo moxibustion therapy was adopted in the NSCs + EPCs moxibustion group, in which, the suspending moxibustion technique was applied to"Baihui"(GV 20), "Dazhui"(GV 14) and "Shenting"(GV 24), 20 min at each acupoint. The treatment was given once every day and a 14-day treatment was as one course. Totally, 3 courses of treatment were required. At the end of treatment, Morris water maze experiment was adopted to determine the learning and memory ability of the rats in each group. In the No.2 experiment, the model rats were divided into 3 groups, a NSCs + EPCs moxibustion group, a NSCs + EPCs blank group and a model group, 18 rats in each one. In each group, according to the durations of treatment,3 subgroups were divided and 6 rats in each one. The intervention method was same as the No.1 experiment. Additionally, after corresponding treatment course, using perfusion, the brains were collected in each subgroup and the slices were frozen. BDNF/TrkB expressions were observed in the immunofluorescence test. Results After treatment
作者
张媛媛
张校军
杨骏
樊吟秋
崔竟成
王频
张庆萍
ZHANG Yuan-yuan;ZHANG Xiao-jun;YANG Jun;FAN Yin-qiu;CUI Jing-cheng;WANG Pin;ZHANG Qing-ping(the First Affiliated Hospital of Anhui University of TCM,Hefei 230031,China;Anhui Integrated Chinese and Western Medicine Hospital;Graduate School of Anhui University of TCM;School of Acupuncture,Moxibustion and Tuina,Anhui University of TCM,Hefei 230031)
出处
《中国针灸》
CAS
CSCD
北大核心
2019年第1期65-71,共7页
Chinese Acupuncture & Moxibustion
基金
国家自然科学基金项目:81473785
