电针对围产期尼古丁暴露子代大鼠记忆和认知损伤影响的机制探讨  

M echanism of effects of electroacupuncture on memory and cognitive impairment in offspring rats with perinatal nicotine exposure

作　　者：张子玥 嵇波[1] 刘翼天 方洋 葛云鹏 谢亚娜 王佳佳 石天宇 Reiko Sakurai Virender Kumar Rehan ZHANG Zi-yue;JI Bo;LIU Yi-tian;FANG Yang;GE Yun-peng;XIE Ya-na;WANG Jia-jia;SHI Tian-yu;Reiko Sakurai;Virender Kumar Rehan(College of Acupuncture-moxibustion and Massage,Beijing University of Chinese Medicine,Beijing 100029,China;Department of Pediatrics,Lundquist Institute for Biomedical Innovation at Harbor-UCLA Medical Center,David Geffen School of Medicine at UCLA,Los Angeles 90001,USA)

机构地区：[1]北京中医药大学针灸推拿学院,北京100029 [2]Harbor医学中心Lundquist生物医学创新研究所儿科系,加州大学洛杉矶分校David Geffen医学院,美国洛杉矶90001

出　　处：《针刺研究》2023年第12期1218-1226,共9页Acupuncture Research

基　　金：国家自然科学基金面上项目(No.81674059、82174505)。

摘　　要：目的:通过观察电针对围产期尼古丁暴露(PNE)诱发子代大鼠宫内生长受限(IUGR)后记忆、认知损伤和脑组织脑源性神经营养因子(BDNF)/N-甲基-D-天冬氨酸受体Ⅰ型(NMDAR1)通路的影响,探讨电针的作用机制。方法:SD大鼠随机分为正常组、模型组、电针组,每组4只母鼠,40只子鼠。采用母鼠孕期哺乳期皮下注射尼古丁制备IUGR模型。从母鼠受孕第6天一直到子代大鼠出生后第21天,电针母鼠双侧“足三里”,每日1次,每次20 min。采用脑脏器系数检测子代大鼠脑发育情况,用Y迷宫和新物体识别实验检测子代大鼠记忆和认知功能,HE染色法观察子代大鼠海马和前额叶皮质发育情况及细胞形态,紫外线比色法检测子代大鼠海马谷氨酸(Glu)含量,ELISA法检测子代大鼠海马BDNF含量,Western blot法检测子代大鼠海马NMDAR1蛋白表达量,免疫组织化学法检测子代大鼠海马和前额叶皮质BDNF阳性细胞数。结果:与正常组比较,模型组子代大鼠脑脏器系数显著降低(P<0.01);新异臂探索时间、自发交替率和新物体识别指数均显著降低(P<0.01);HE染色示海马和前额叶皮质的神经元数量减少,排列散乱,细胞整体结构遭到破坏;海马中Glu蛋白含量显著升高(P<0.01),BDNF蛋白含量和NMDAR1蛋白表达量均显著降低(P<0.01);海马CA1区、CA3区和前额叶皮质BDNF阳性细胞数减少(P<0.01)。与模型组比较,电针组子代大鼠脑脏器系数显著升高(P<0.01);新异臂探索时间、自发交替率和新物体识别指数均显著升高(P<0.05,P<0.01);HE染色示海马和前额叶皮质的神经元数量增多,排列整齐,细胞整体结构损伤减轻;海马中Glu蛋白含量显著降低(P<0.01),BDNF蛋白含量和NMDAR1蛋白表达量均显著升高(P<0.05);海马CA1区、CA3区和前额叶皮质BDNF阳性细胞数增多(P<0.05,P<0.01)。结论:电针对PNE所导致的IUGR子代大鼠记忆和认知功能损伤具有改善作用,其机制可能与通过调节BDObjectiveTo observe the effects of electroacupuncture(EA)on m em ory,cognitive im pairm ent,and the brain-derived neurotrophic factor(BDNF)/N-m ethyl-D-aspartate receptor subtype 1(NM DAR1)pathway in the brains of offspring rat with intrauterine growth restriction(IUGR)induced by perinatal nicotine exposure(PNE),so as to explore the underlying m echanism.MethodsSD rats were random ly divided into norm al,m odel,and EA groups,with 4 m others and 10 offspring rats of each m other in each group.The IUGR m odel was established by subcutaneous injection of nicotine during pregnancy and lactation.From the 6th day of pregnancy in the m others until the 21st day after birth of the offspring rats,EA(2 Hz/15 Hz,1 m A)was adm inistered bilaterally at the“Zusanli”(ST36)of m others,once daily for 20 m in.The brain organ coefficient was used to evaluate the brain developm ent of the offspring rats.The Y-m aze test and novel object recognition experim ents were perform ed to assess m em ory and cognitive function.HE staining was used to observe the developm ent and cellular m orphology of the hippocam pus and prefrontal cortex in the offspring rats.UV spectrophotom etry was used to m easure the glutam ate(Glu)content in the hippocam pus.ELISA was used to detect the BDNF content in the hippocam pus.W estern blot was perform ed to m easure the protein expression of NM DAR1 in the hippocam pus.Im m unohistochem istry was used to count the num ber of BDNF-positive cells in the hippo⁃cam pus and prefrontal cortex.ResultsCom pared with the norm al group,the brain organ coefficient,exploration tim e of the novel arm,spontaneous alternation rate,and novel object recognition index,contents of BDNF and expression of NM DAR1 proteins in the hippocam pus,the num ber of BDNF-positive cells in the CA1 and CA3 regions of the hippo⁃cam pus and prefrontal cortex were significantly reduced(P<0.01),while the Glu content in the hippocam pus was signifi⁃cantly increased(P<0.01)in the m odel group of offspring rats;decreased cell num ber,s

关 键 词：电针 宫内生长受限 围产期 尼古丁暴露 认知记忆 足三里 海马 前额叶皮质 

分 类 号：R245.97[医药卫生—针灸推拿学]