主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2021, Vol. 44 ›› Issue (4): 323-331.doi: 10.3969/j.issn.1006-2157.2021.04.007

• 中药药理 • 上一篇    下一篇

基于BDNF/TrkB介导的海马突触可塑性研究补肝散对衰老大鼠学习记忆功能的影响*

王硕1, 李伟1, 王浩1,2, 王杰鹏1,2, 方芳1,2, 王少贤1,2, 王建1, 方朝义1,2#   

  1. 1 河北中医学院 河北 050200;
    2 河北省中西医结合肺病研究重点实验室
  • 收稿日期:2020-09-28 发布日期:2021-04-29
  • 通讯作者: #方朝义,男,博士,教授,博士生导师,主要研究方向:衰老疾病中医证治规律研究,E-mail:hbfcy@163.com
  • 作者简介:王硕,女,在读硕士生
  • 基金资助:
    *国家自然科学基金项目(No.81673881),河北省中医药类科研计划项目(No.2020136),河北中医学院省属高校基本科研业务费专项项目(No.YJZ2019006),河北中医学院研究生创新资助项目(No.XCXZZSS2020009)

Effects of Bugansan on learning and memory function of aging rats based on BDNF/TrkB mediated hippocampal synaptic plasticity*

Wang Shuo1, Li Wei1, Wang Hao1,2, Wang Jiepeng1,2, Fang Fang1,2, Wang Shaoxian1,2, Wang Jian1, Fang Chaoyi1,2#   

  1. 1 Hebei University of Traditional Chinese Medicine, Hebei 050200, China;
    2 Hebei Key Laboratory of Integrated Chinese and Western Medicine for Lung Disease Research, Hebei 050091, China
  • Received:2020-09-28 Published:2021-04-29
  • Contact: Prof. Fang Chaoyi, Doctoral Supervisor. School of Basic Medicine, Hebei University of Traditional Chinese Medicine, 3 Xingyuan Road, Luquan District, Shijiazhuang 050200. E-mail: hbfcy@163.com
  • Supported by:
    National Natural Science Foundation of China (No.81673881); Traditional Chinese Medicine Scientific Research Project of Heibei Province(No.2020136); Hebei University of Traditional Chinese Medicine Provincial Universities Basic Scientific Research Operating Expenses Special Project(No.YJZ2019006); Postgraduate Innovation Support Project of Hebei University of Traditional Chinese Medicine (No.XCXZZSS2020009)
    Ethical review: Ethics Committee of Hebei University of Traditional Chinese Medicine (No.DWLL2018040)

摘要: 目的 基于肝气虚理论和海马突触可塑性探讨衰老机制,观察补肝散对D-半乳糖致衰老大鼠学习记忆能力的影响。方法 40只SPF级SD雄性大鼠随机分为空白对照组、模型组、吡拉西坦组和补肝散组,各10只。模型组、吡拉西坦组、补肝散组腹腔注射400 mg/kg D-半乳糖制备衰老大鼠模型,空白对照组腹腔注射等量生理盐水,每日1次,连续8周。造模当日始,补肝散组予以补肝散煎液灌胃(14.06 g/kg),吡拉西坦组予以吡拉西坦混悬液灌胃(0.40 g/kg),空白对照组和模型组给予等量蒸馏水(10 mL/kg),每日1次,连续8周。取材前1天称量大鼠体质量;抓力实验检测大鼠抓力大小;八臂迷宫和跳台实验评估大鼠学习记忆能力;尼氏染色观察海马神经元病理变化;免疫组织化学染色观察海马突触素(SYN)和突触后致密蛋白95(PSD95)表达特征;实时荧光定量PCR和免疫印迹法(Western blotting)分别检测海马脑源性神经营养因子(BDNF)、原肌球蛋白相关激酶B(TrkB)基因和蛋白表达。结果 与空白对照组比较,模型组大鼠体质量下降,抓力降低,工作记忆错误、参考记忆错误和总记忆错误次数明显升高,平台潜伏期缩短,海马神经元数量明显减少,SYN和PSD95表达水平显著下降,BDNF、TrkB基因和蛋白表达水平显著降低(均P<0.05);与模型组比较,补肝散组大鼠体质量增加,抓力增大,工作记忆错误、参考记忆错误和总记忆错误次数显著下降,平台潜伏期延长,海马神经元数量显著增加,SYN和PSD95表达水平升高,BDNF、TrkB基因和蛋白表达水平显著升高(均P<0.05)。结论 肝气亏虚是机体衰老的重要机制,调补肝气为其有效治法;补肝散可通过上调BDNF和TrkB水平以增强衰老大鼠海马突触可塑性,从而改善学习记忆能力,达到延缓衰老的目的。

关键词: 衰老, 肝气亏虚, 海马突触可塑性, BDNF/TrkB信号通路, 学习记忆, 补肝散, 大鼠

Abstract: Objective To explore the aging mechanism based on the theory of liver qi deficiency and hippocampal synaptic plasticity, and to observe the effects of Bugansan (Liver-tonifying Powder) formula on the learning and memory abilities of D-galactose induced aging rats. Methods Forty SPF male SD rats were randomly divided into blank group, model group, piracetam group and Bugansan group (n=10 in each group). Rats in the model group, piracetam group and Bugansan group were intraperitoneally injected with 400 mg/kg D-galactose to establish aging rat models, while blank control group was intraperitoneally injected with the same amount of normal saline daily for eight consecutive weeks. From the day of modeling, Bugansan group was given Bugansan decoction (14.06 g/kg), while piracetam group was given piracetam suspension (0.40 g/kg); blank control group and model group were given the same amount of distilled water (10 mL/kg) daily for 8 consecutive weeks. The body weight of the rats was measured the day before sampling; grip strength test was used to detect the grip strength of rats; Eight-arm maze and step-down test were used to evaluate the learning and memory abilities of rats; Nissl staining method was used to measure the pathological changes of hippocampal neurons while immunohistochemical staining was used to assess the expression characteristics of synaptophysin (SYN) and postsynaptic density 95 (PSD95); Real-time quantitative PCR and Western blotting were used to detect the expression of brain-derived neurotrophic factor (BDNF), tropomyosin-related kinase B (TrkB) genes and proteins in hippocampus. Results Compared with the blank group, body weight and grip strength of rats in model group decreased. The number of working memory error (WME), reference memory error (RME) and total memory errors (TE) were significantly increased. The platform latency was shortened; the number of hippocampal neurons was significantly reduced. The expression levels of SYN and PSD95 were significantly decreased, and the expression levels of BDNF and TrkB genes and proteins were significantly decreased (all P<0.05). Compared with the model group, the body weight and grip strength of rats in Bugansan group increased. The number of WME, RME and TE were significantly decreased. The platform latency was prolonged. The number of hippocampal neurons was significantly increased. The expression of SYN and PSD95 was increased, and the expression levels of BDNF and TrkB genes and proteins were significantly increased (all P<0.05). Conclusion Deficiency of liver qi is an important mechanism of body aging. Regulating and replenishing liver qi is an effective treatment. Bugansan can enhance synaptic plasticity in hippocampus of aging rats by up-regulating BDNF and TrkB levels, thereby improving learning and memory abilities and achieving the purpose of delaying aging.

Key words: aging, liver qi deficiency, hippocampal synaptic plasticity, BDNF/TrkB signaling pathway, learning and memory, Bugansan, rats

中图分类号: 

  • R285.5