Electroacupuncture Reduced Apoptosis of Hippocampal Neurons in Mice with Cerebral Infarction by Regulating the Notch3 Signaling Pathway

  • Ruisi Tian
  • Shu WangEmail author


This study is designed to explore the effect of electric acupuncture on the apoptosis of hippocampal neurons through the Notch3 signaling pathway. A middle cerebral artery occlusion (MCAO) mouse model was generated by thread embolization. A Y-maze was used to test cognitive function (learning and memory). The apoptosis factor caspase-3 was identified through an enzyme-linked immunosorbent assay (ELISA). TTC staining was used to evaluate the cerebral infarction area. Apoptosis of hippocampal neurons was identified by TUNEL staining and flow cytometry. The expression levels of genes and proteins related to the Notch3 signaling pathway were assessed by qRT-PCR and Western blot. It was determined that 7 days of electroacupuncture treatment improved recovery from cerebral infarction. Electroacupuncture therapy reduced the cerebral infarction area in mice and decreased the apoptosis of hippocampal neurons. The apoptosis rate of hippocampal neurons and the area of cerebral infarction were significantly lower in the electroacupuncture group than in the MCAO group (P < 0.01), while the apoptosis rate of hippocampal neurons and the area of cerebral infarction were significantly higher in the inhibitor group than in the electroacupuncture group (P < 0.01). The expression levels of genes related to the Notch3 signaling pathway were significantly increased in the electric acupuncture treatment group compared with the MCAO group (P < 0.01). The results showed that electroacupuncture inhibited the apoptosis of hippocampal neurons through the Notch3 signaling pathway, while the expression levels of genes related to the Notch3 signaling pathway were significantly higher in the inhibitor group than in the electroacupuncture group (P < 0.01). Electroacupuncture inhibited the apoptosis of hippocampal neurons in mice with cerebral infarction by stimulating the Notch3 signaling pathway and triggering corresponding protein expression.


Cerebral infarction Electric acupuncture Notch3 signaling pathway Hippocampal neuron 


Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine.

Informed Consent

Informed consent was obtained from all individual participants included in the study.


  1. Ates Sonmezoglu O, Terzi B (2018) Characterization of some bread wheat genotypes using molecular markers for drought tolerance. Physiol Mol Biol Plants 24:159–166. CrossRefPubMedGoogle Scholar
  2. Broughton BR, Reutens DC, Sobey CG (2009) Apoptotic mechanisms after cerebral ischemia. Stroke 40:e331–e339. CrossRefPubMedGoogle Scholar
  3. Campbell MT, Dagher P, Hile KL, Zhang H, Meldrum DR, Rink RC, Meldrum KK (2008) Tumor necrosis factor-alpha induces intrinsic apoptotic signaling during renal obstruction through truncated bid activation. J Urol 180:2694–2700. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Caplan LR, Searls DE, Hon FK (2009) Cerebrovascular disease. Med Clin North Am 93:353–369, viii. CrossRefPubMedGoogle Scholar
  5. Chang QY, Lin YW, Hsieh CL (2018) Acupuncture and neuroregeneration in ischemic stroke. Neural Regen Res 13:573–583. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Cui S, Xu M, Huang J et al (2018) Cerebral responses to acupuncture at GV24 and bilateral GB13 in rat models of Alzheimer's disease. Behav Neurol 2018:8740284. CrossRefPubMedPubMedCentralGoogle Scholar
  7. D'Angelo RC, Ouzounova M, Davis A, Choi D, Tchuenkam SM, Kim G, Luther T, Quraishi AA, Senbabaoglu Y, Conley SJ, Clouthier SG, Hassan KA, Wicha MS, Korkaya H (2015) Notch reporter activity in breast cancer cell lines identifies a subset of cells with stem cell activity. Mol Cancer Ther 14:779–787. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Ding W, Chen R, Wu C, Chen W, Zhang H, Fan X, Wang H, Ji Y, Xie L, Ning X, Shen L (2017) Increased expression of HERPUD1 involves in neuronal apoptosis after intracerebral hemorrhage. Brain Res Bull 128:40–47. CrossRefPubMedGoogle Scholar
  9. Du Y, Shi L, Li J, Xiong J, Li B, Fan X (2011) Angiogenesis and improved cerebral blood flow in the ischemic boundary area were detected after electroacupuncture treatment to rats with ischemic stroke. Neurol Res 33:101–107. CrossRefPubMedGoogle Scholar
  10. Ehret F, Vogler S, Pojar S, Elliott DA, Bradke F, Steiner B, Kempermann G (2015) Mouse model of CADASIL reveals novel insights into Notch3 function in adult hippocampal neurogenesis. Neurobiol Dis 75:131–141. CrossRefPubMedGoogle Scholar
  11. Jiang Q, Li RP, Tang Y, Wang YQ, Liu C, Guo ML (2015) Bakkenolide-IIIa protects against cerebral damage via inhibiting NF-kappaB activation. CNS Neurosci Ther 21:943–952. CrossRefPubMedGoogle Scholar
  12. Kim YR, Kim HN, Jang JY et al (2013) Effects of electroacupuncture on apoptotic pathways in a rat model of focal cerebral ischemia. Int J Mol Med 32:1303–1310. CrossRefPubMedGoogle Scholar
  13. Kopan R, Ilagan MX (2009) The canonical Notch signaling pathway: unfolding the activation mechanism. Cell 137:216–233. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Li X, Luo P, Wang Q, Xiong L (2012) Electroacupuncture pretreatment as a novel avenue to protect brain against ischemia and reperfusion injury. Evid Based Complement Alternat Med 2012:195397. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Liu J, Wang Q, Yang S, Huang J, Feng X, Peng J, Lin Z, Liu W, Tao J, Chen L (2018) Electroacupuncture inhibits apoptosis of peri-ischemic regions via modulating p38, extracellular signal-regulated kinase (ERK1/2), and c-Jun N terminal kinases (JNK) in cerebral ischemia-reperfusion-injured rats. Med Sci Monit 24:4395–4404. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Lu ZQ, Deng YJ, Lu JX (2012) Effect of aloe polysaccharide on caspase-3 expression following cerebral ischemia and reperfusion injury in rats. Mol Med Rep 6:371–374. CrossRefPubMedGoogle Scholar
  17. Lugert S, Basak O, Knuckles P, Haussler U, Fabel K, Götz M, Haas CA, Kempermann G, Taylor V, Giachino C (2010) Quiescent and active hippocampal neural stem cells with distinct morphologies respond selectively to physiological and pathological stimuli and aging. Cell Stem Cell 6:445–456. CrossRefPubMedGoogle Scholar
  18. Mu S, Liu B, Ouyang L, Zhan M, Chen S, Wu J, Chen J, Wei X, Wang W, Zhang J, Lei W (2016) Characteristic changes of astrocyte and microglia in rat striatum induced by 3-NP and MCAO. Neurochem Res 41:707–714. CrossRefPubMedGoogle Scholar
  19. Nakka VP, Gusain A, Mehta SL, Raghubir R (2008) Molecular mechanisms of apoptosis in cerebral ischemia: multiple neuroprotective opportunities. Mol Neurobiol 37:7–38. CrossRefPubMedGoogle Scholar
  20. Qi L, Han Z, Zhou Y, Chen W, Chu L, Lu J, Xu W, Wang H, Wang Z, Ling J (2018) Dynamic scalp acupuncture combined with PNF therapy for upper limb motor impairment in ischemic stroke spastic hemiplegia. Zhongguo Zhen Jiu 38:234–238. CrossRefPubMedGoogle Scholar
  21. She YF, Qi CH, Zhu J (2010) History and progress of study on electrical properties of acupoints at home and abroad. Zhongguo Zhen Jiu 30:1047–1050PubMedGoogle Scholar
  22. Wang Z, Da Silva TG, Jin K et al (2014a) Notch signaling drives stemness and tumorigenicity of esophageal adenocarcinoma. Cancer Res 74:6364–6374. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Wang Y, Wan C, Yu S, Yang L, Li B, Lu T, Bi Y, Jiang J, Cui G (2014b) Upregulated expression of NF-YC contributes to neuronal apoptosis via proapoptotic protein bim in rats’ brain hippocampus following middle cerebral artery occlusion (MCAO). J Mol Neurosci 52:552–565. CrossRefPubMedGoogle Scholar
  24. Wang C, Wu Z, Li N, Zhao Y, Tian F, Zhou X, Wang Z (2014c) Clinical curative effect of electric acupuncture on acute cerebral infarction: a randomized controlled multicenter trial. J Tradit Chin Med 34:635–640CrossRefGoogle Scholar
  25. Yin J, Li H, Feng C, Zuo Z (2014) Inhibition of brain ischemia-caused notch activation in microglia may contribute to isoflurane postconditioning-induced neuroprotection in male rats. CNS Neurol Disord Drug Targets 13:718–732CrossRefGoogle Scholar
  26. Yuan X, Wu H, Han N, Xu H, Chu Q, Yu S, Chen Y, Wu K (2014) Notch signaling and EMT in non-small cell lung cancer: biological significance and therapeutic application. J Hematol Oncol 7:87. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Zhang X, Huang G, Liu H, Chang H, Wilson JX (2012) Folic acid enhances Notch signaling, hippocampal neurogenesis, and cognitive function in a rat model of cerebral ischemia. Nutr Neurosci 15:55–61. CrossRefPubMedGoogle Scholar
  28. Zhao Y, Chen X, Ma L, Zuo Z, Zhu Z, Zhu X, Wang Q, He E, Xiong L, Pei J, Xu L, Hou L, Chen S (2012) Electroacupuncture pretreatment induces tolerance against focal cerebral ischemia through activation of canonical Notch pathway. BMC Neurosci 13:111. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Zhou Y, Tong D, Wang S, Liu L, Ye S, Xu B (2017a) Comparison of risk of death between older and non-older critical patients in ICU: a retrospective cohort study of consecutive 3 years. Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 29:448–452. CrossRefPubMedGoogle Scholar
  30. Zhou L, Zhang J, Wang C, Sun Q (2017b) Tanshinone inhibits neuronal cell apoptosis and inflammatory response in cerebral infarction rat model. Int J Immunopathol Pharmacol 30:123–129. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Zhuang Y, Xing JJ, Li J, Zeng BY, Liang FR (2013) History of acupuncture research. Int Rev Neurobiol 111:1–23. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Rehabilitation Medicine DepartmentThe Second Affiliated Hospital of Xinjiang Medical UniversityUrumqiChina
  2. 2.Institute of Acupuncture and MoxibustionThe First Teaching Hospital of Tianjin University of Traditional Chinese MedicineNankai DistrictChina

Personalised recommendations