The Effect of Electroacupuncture on Neuronal Apoptosis and Related Functions in Rats with Acute Spinal Cord Injury

Liang Zhang¹, Changming Li¹, Renfu Quan^2*, Shangju Xie¹

¹Research Institute of Orthopedics, Zhejiang Chinese Medical University, Hangzhou, China.
²Department of Orthopedics, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, China.

DOI: 10.4236/cm.2014.54024                

Abstract

Objective: To investigate the effect and significance of electroacupuncture (EA) on neuronal apoptosis and hindlimb motor and bladder functional improvement in rats with acute spinal cord injury (SCI). Methods: Sixty healthy Sprague Dawley rats were randomly assigned to sham, model, EA, and EA control groups (n = 15 each). EA group rats received EA treatment at Zhibian and Shuidao acupoints seven times daily, whereas EA control group rats received EA at two points, 0.5 cm away from Zhibian and Shuidao, respectively. Histomorphological changes in spinal cord tissue were examined using hematoxylin-eosin staining. Neuronal apoptosis was detected by TUNEL assay. Bcl-2, Bax, and Bad protein levels were detected using immunohistochemistry. Additionally, hindlimb motor function, residual urine volume and maximum bladder capacity were measured. Results: HE staining revealed a morphologically and structurally intact spinal cord in the EA group, and the tissue contained scattered blood cells without edema. In the EA control group, there were small morphological defects in the spinal cord, and the tissue contained fewer blood cells with local edema. Compared with the EA control and model groups, Bax and Bad levels were significantly decreased in the EA group and Bcl-2 expression was increased (P < 0.05). After SCI, hindlimb function scores, residual urine volume, and maximum bladder capacity in rats of the EA group significantly differed from those of the EA control group (P < 0.05). Conclusion: EA may induce SCI-induced improvements in hindlimb motor and bladder functions by affecting neuronal apoptosis and relevant gene expression changes.

Keywords

Electroacupuncture, Spinal Cord Injury, Neuronal Apoptosis, Molecular Mechanism

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Xia, L.P., Fan, F., Tang, A.L. and Ye, W.Q. (2014) Effects of Electroacupuncture Combined with Bladder Training on the Bladder Function of Patients with Neurogenic Bladder after Spinal Cord Injury. International Journal of Clinical and Experimental Medicine, 7, 1344-1348.
[2]	Fischer, M.J., Krishnamoorthi, V.R., Smith, B.M., Evans, C.T., St Andre, J.R., Ganesh, S., et al. (2012) Prevalence of Chronic Kidney Disease in Patients with Spinal Cord Injuries/Disorders. American Journal of Nephrology, 36, 542548. http://dx.doi.org/10.1159/000345460
[3]	Xing, G.G., Liu, F.Y., Qu, X.X., Han, J.S. and Wan, Y. (2007) Long-Term Synaptic Plasticity in the Spinal Dorsal Horn and Its Modulation by Electroacupuncture in Rats with Neuropathic Pain. Experimental Neurology, 208, 323-332. http://dx.doi.org/10.1016/j.expneurol.2007.09.004
[4]	Hwang, L., Choi, I.Y., Kim, S.E., Ko, I.G., Shin, M.S., Kim, C.J., et al. (2013) Dexmedetomidine Ameliorates Intracerebral Hemorrhage-Induced Memory Impairment by Inhibiting Apoptosis Andenhancing Brain-Derived Neurotrophic Factor Expression in the Rat Hippocampus. International Journal of Molecular Medicine, 31, 1047-1056.
[5]	Sung, Y.H., Kim, S.C., Hong, H.P., Park, C.Y., Shin, M.S., Kim, C.J., et al. (2012) Treadmill Exercise Ameliorates Dopaminergic Neuronal Loss through Suppressing Microglial Activation in Parkinson’s Disease Mice. Life Sciences, 91, 1309-1316. http://dx.doi.org/10.1016/j.lfs.2012.10.003
[6]	Okuno, S., Saito, A., Hayashi, T. and Chan, P.H. (2004) The c-Jun N-Terminal Protein Kinase Signaling Pathway Mediates Bax Activation and Subsequent Neuronal Apoptosis through Interaction with Bim after Transient Focal Cerebral Ischemia. Journal of Neuroscience, 24, 7879-7887. http://dx.doi.org/10.1523/JNEUROSCI.1745-04.2004
[7]	Chen, M.H., Ren, Q.X., Yang, W.F., Chen, X.L., Lu, C. and Sun, J. (2013) Influences of HIF-lα on Bax/Bcl-2 and VEGF Expressions in Rats with Spinal Cord Injury. International Journal of Clinical and Experimental Pathology, 6, 2312-2322.
[8]	Li, Y., Gu, J., Liu, Y., Long, H., Wang, G., Yin, G., et al. (2013) iNOS Participates in Apoptosis of Spinal Cord Neurons via p-BAD Dephosphorylation Following Ischemia/Reperfusion (I/R) Injury in Rat Spinal Cord. Neuroscience Letters, 545, 117-122. http://dx.doi.org/10.1016/j.neulet.2013.04.043
[9]	Sun, W., Winseck, A., Vinsant, S., Park, O.H., Kim, H. and Oppenheim, R.W. (2004) Programmed Cell Death of Adult-Generated Hippocampal Neurons Is Mediated by the Proapoptotic Gene Bax. Journal of Neuroscience, 24, 11205-11213. http://dx.doi.org/10.1523/JNEUROSCI.1436-04.2004
[10]	Norrbrink, C. and Lundeberg, T. (2011) Acupuncture and Massage Therapy for Neuropathic Pain Following Spinal Cord Injury: An Exploratory Study. Acupuncture in Medicine, 29, 108-115. http://dx.doi.org/10.1136/aim.2010.003269
[11]	Pei, J., Sun, L., Chen, R., Zhu, T., Qian, Y. and Yuan, D. (2001) The Effect of Electro-Acupuncture on Motor Function Recovery in Patients with Acute Cerebral Infarction: A Randomly Controlled Trial. Journal of Traditional Chinese Medicine, 21, 270-272.
[12]	Lin, H.S., Ji, Z.S., Zheng, L.H., Guo, G.Q., Chen, B., Zhang, G.W. and Wu, H. (2012) Effect of Methylprednisolone on the Activities of Caspase-3, -6, -8 and -9 in Rabbits with Acute Spinal Cord Injury. Experimental and Therapeutic Medicine, 4, 49-54.
[13]	Zhou, Y., Liu, X.H., Qu, S.D., Yang, J., Wang, Z.W., Gao, C.J., et al. (2013) Hyperbaric Oxygen Intervention on Expression of Hypoxia-Inducible Factor-1α and Vascular Endothelial Growth Factor in Spinal Cord Injury Models in Rats. Chinese Medical Journal (English), 126, 3897-3903.
[14]	Rajan, R.K., Juler, G. and Eltorai, I.M. (2000) Sigmoid Colon Rupture Secondary to Credé’s Method in a Patient with Spinal Cord Injury. The Journal of Spinal Cord Medicine, 23, 90-91.
[15]	Nomura, S., Ishido, T., Teranishi, J. and Makiyama, K. (2000) Long-Term Analysis of Suprapubic Cystostomy Drainage in Patients with Neurogenic Bladder. Urologia Internationalis, 65, 185-189. http://dx.doi.org/10.1159/000064873
[16]	Feng, X.D., Yang, S.L., Liu, J., Huang, J., Peng, J., Lin, J.M., et al. (2013) Electroacupuncture Ameliorates Cognitive Impairment through Inhibition of NF-κB-Mediated Neuronal Cell Apoptosis in Cerebral Ischemia-Reperfusion Injured Rats. Molecular Medicine Reports, 7, 1516-1522.
[17]	Lee, B., Sur, B., Shim, J., Hahm, D.H. and Lee, H. (2014) Acupuncture Stimulation Improves Scopolamine-Induced Cognitive Impairment via Activation of Cholinergic System and Regulation of BDNF and CREB Expressions in Rats. BMC Complementary and Alternative Medicine, 14, 338.
[18]	Cha, M.H., Bai, S.J., Lee, K.H., Cho, Z.H., Kim, Y.B., Lee, H.J., et al. (2010) Acute Electroacupuncture Inhibits Nitric oxide Synthase Expression in the Spinal Cord of Neuropathic Rats. Neurological Research, 32, 96-100. http://dx.doi.org/10.1179/016164109X12537002794363
[19]	Liu, Z., Ding, Y. and Zeng, Y.S. (2011) A New Combined Therapeutic Strategy of Governor Vessel Electro-Acupuncture and Adult Stem Cell Transplantation Promotes the Recovery of Injured Spinal Cord. Current Medicinal Chemistry, 18, 5165-5171. http://dx.doi.org/10.2174/092986711797636144
[20]	Kim, D.H., Ko, I.G., Kim, B.K., Kim, T.W., Kim, S.E., Shin, M.S., et al. (2010) Treadmill Exercise Inhibits Traumatic Brain Injury-Induced Hippocampal Apoptosis. Physiology & Behavior, 101, 660-665. http://dx.doi.org/10.1016/j.physbeh.2010.09.021
[21]	Crowe, M.J., Bresnahan, J.C., Shuman, S.L., Masters, J.N. and Beattie, M.S. (1997) Apoptosis and Delayed Degeneration after Spinal Cord Injury in Rats and Monkeys. Nature Medicine, 3, 73-76.
[22]	Hou, Q., Cymbalyuk, E., Hsu, S.C., Xu, M. and Hsu, Y.T. (2003) Apoptosis Modulatory Activities of Transiently Expressed Bcl-2: Roles in Cytochrome c Release and Bax Regulation. Apoptosis, 8, 617-629. http://dx.doi.org/10.1023/A:1026187526113
[23]	Zhang, X.H., Chen, S.Y., Tang, L., Shen, Y.Z., Luo, L., Xu, C.W., et al. (2013) Myricetin Induces Apoptosis in HepG2 Cells through Akt/p70S6K/Bad Signaling and Mitochondrial Apoptotic Pathway. Anti-Cancer Agents in Medicinal Chemistry, 13, 1575-1581. http://dx.doi.org/10.2174/1871520613666131125123059
[24]	Bleicken, S., Wagner, C. and Garcia-Saez, A.J. (2013) Mechanistic Differences in the Membrane Activity of Bax and Bcl-xL Correlate with Their Opposing Roles in Apoptosis. Biophysical Journal, 104, 421-431. http://dx.doi.org/10.1016/j.bpj.2012.12.010
[25]	Wang, T., Liu, C.Z., Yu, J.C., Jiang, W. and Han, J.X. (2009) Acupuncture Protected Cerebral Multi-Infarction Rats from Memory Impairment by Regulating the Expression of Apoptosis Related Genes Bcl-2 and Bax in Hippocampus. Physiology & Behavior, 96, 155-161. http://dx.doi.org/10.1016/j.physbeh.2008.09.024