基于BDNF/CREB信号通路探讨巴戟天对慢性应激抑郁大鼠海马神经元损伤的影响  

Effect of Bajitian(Morindae Officinalis Radix)on Hippocampal Neurons Injury of Chronic Stress Depression Rats Based on BDNF/CREB Signaling Pathway

作　　者：王钦 刁丽梅[2] 蔡萧君 WANG Qin;DIAO Limei;CAI Xiaojun(Guangxi University of Chinese Medicine,Nanning 530200,Guangxi,China;The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China;Traditional Chinese Medicine Academy of Heilongjiang,Harbin 150000,Heilongjiang,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]广西中医药大学第一附属医院,广西南宁530023 [3]黑龙江省中医药科学院,黑龙江哈尔滨150000

出　　处：《中华中医药学刊》2024年第2期69-74,I0017,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81960858);广西中医脑病临床研究中心项目(桂科AD20238028);广西科技基地和人才专项(桂科AD22035168);广西高等学校高水平创新团队及卓越学者计划项目(桂教人才[2020]6号);广西中医药大学第一附属医院学术团队建设项目(院字[2018]146号);广西中医药大学研究生教育创新计划项目(YCBXJ2023005)。

摘　　要：目的研究巴戟天对慢性应激抑郁大鼠海马神经元损伤及脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)/细胞内环磷腺苷效应元件结合蛋白(cyclic adenosine phosphate response element binding protein,CREB)信号通路的影响。方法从40只SD大鼠随机选取其中10只为正常组,对其余大鼠构建慢性不可预知温和应激模型后,再将其分为3组,即模型组,盐酸氟西汀组(3.17 mg·kg^(-1)),巴戟天组(3.17 g·kg^(-1))。持续灌胃后给药8周,1次/d,并先后在造模前、造模后和给药后开展了行为学实验,以判断大鼠的抑郁情况。通过苏木素-伊红染色(hematoxylin-eosin staining,HE)研究大鼠海马形态学改变,用免疫组织化学法(Immunohistochemistry,IHC)测定大鼠海马BDNF蛋白表达,用HE染色研究大鼠海马组织病理损伤并评分,用实时荧光定量聚合酶链式反应(Real-time PCR,RT-PCR)测定大鼠海马BDNF、TrkB、CREB mRNA相对表达,用蛋白免疫印迹法(western blot,WB)测定大鼠海马BDNF、TrkB、CREB蛋白的相对表达。结果与正常组比较,模型组大鼠活动总路程显著减少(P<0.05),自主游泳时间显著减少(P<0.05),悬尾挣扎时间显著减少(P<0.05)。HE染色结果中表明海马神经元组织受到破坏,病理损伤评分显著下降(P<0.05),免疫组织化学染色中海马BDNF表现显著下降(P<0.05),BDNF、TrkB、CREB mRNA的基因相对表达显著下降(P<0.05);与模型组对比,盐酸氟西汀组和巴戟天组大鼠活动的总里程明显提高(P<0.05),自主游泳时间显著增加(P<0.05),悬尾挣扎时间显著增加(P<0.05),HE染色结果表明海马神经元组织明显复原,病理损伤评分增加(P<0.05),免疫组织化学染色中BDNF表现显著增加(P<0.05),BDNF、TrkB、CREB mRNA的基因相对表达明显增加(P<0.05)。结论经慢性应激刺激后,巴戟天可能通过激活BDNF/TrkB/CREB信号通路对抑郁大鼠海马神经元损伤发挥神经保护作用,改善其抑郁样行为。Objective To investigate the effects of Bajitian(Morindae Officinalis Radix)on hippocampal neuronal injury and brain-derived neurotrophic factor(BDNF)/cyclic adenosine phosphos effter-element binding protein(CREB)signaling pathway in rats with chronic stress depression.Methods A total of 40 SD rats were randomly selected as normal group(n=10),and the other rats were randomly divided into 3 groups:model group,fluoxetine hydrochloride group(3.17 mg·kg^(-1))and Bajitian(Morindae Officinalis Radix)group(3.17 g·kg^(-1)).The rats were administered orally for 8 weeks,once a day.Behavioral tests were performed before and after modeling and after the rats were administered.Hippocampal morphological changes were observed by hematoxylin-eosin(HE)staining,the expression of hippocampal BDNF protein was detected by immunohistochemistry(IHC),the pathological damage of hippocampal tissue was observed and scored by HE staining,and the levels of BDNF,TrkB and TRKB were detected by Real-time polymerase chain reaction(PCR).The relative expression of CREB mRNA was detected by Western blot.The relative expressions of BDNF,TrkB and CREB protein were detected by Western blot.Results Compared with those of the normal group,the total activity distance of the model group was significantly decreased(P<0.05),the autonomous swimming time was significantly decreased(P<0.05),and the tail hanging and struggling time was significantly decreased(P<0.05).HE staining showed the hippocampal neuronal structure was damaged and the pathological injury score was obviously decreased(P<0.05),the expression of BDNF was significantly decreased(P<0.05),and the relative expressions of BDNF,TrkB,CREB mRNA and protein were significantly decreased(P<0.05).Compared with those of the model group,the total activity distance of rats in fluoxetine hydrochloride group and Bajitian(Morindae Officinalis Radix)group was significantly increased(P<0.05),the autonomous swimming time was significantly increased(P<0.05),and the tail hanging struggle time was significantly in

关 键 词：巴戟天 抑郁 海马 脑源性神经营养因子(BDNF)/环磷腺苷效应元件结合蛋白(CREB) 机制研究 

分 类 号：R285.5[医药卫生—中药学]