Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (4): 416-423.doi: 10.3736/jcim20120410

Previous Articles     Next Articles

Effects of electroacupuncture on expression of c-fos protein and N-methyl-D-aspartate receptor 1 in the rostral ventromedia medulla of rats with chronic visceral hyperalgesia

De-bo Qi, Wei-min Li()   

  1. Laboratory of Neural Network and Systems Biology, Shanghai Medical College, Fudan University, Shanghai 200032, China
  • Received:2011-12-05 Accepted:2011-12-27 Online:2012-04-20 Published:2018-04-15

Objective: Acupuncture has been clinically proved to be effective in treating abdominal pain in patients with irritable bowel syndrome (IBS). However, its neurobiological mechanism remains largely unexplored. The aim of this study was to investigate the effect of electroacupuncture (EA) in relieving chronic visceral hyperalgesia and the possible involvement of N-methyl-D-aspartate receptor 1 (NR1) in rostral ventromedia medulla (RVM) of the brain in an IBS rat model.
Methods: To establish the IBS rat model, male Sprague-Dawley neonatal rats received colon mechanical irritation on a daily basis from the 9th to the 22nd day after their birth. After a resting period of another two to four weeks, behavioral tests of pain threshold pressure (PTP) and abdominal withdrawal reflex (AWR) responding to colorectal distention (CRD) stimuli were conducted to judge the colorectal sensitive situation. Then administration of EA at acupoints of Zusanli (ST36) and Shangjuxu (ST37) bilaterally in the hind limbs was repeated four times every other day, while sham-EA was done by inserting needles at similar acupoints without electrical stimulation. Immunohistochemical method was used to display the expression of proto-oncogene protein c-fos and NR1 in RVM of rats.
Results: The results demonstrated that the PTP values and AWR scores, in response to the CRD stimuli, significantly decreased and increased, respectively (P<0.01, P<0.01), while the number of immunoreactive neurons of c-fos protein and NR1 significantly increased in nucleus reticularis gigantocellularis (Gi), nucleus lateralis paragigantocellulari (LPGi), nucleus reticularis gigantocellularis pars alpha (GiA) and nucleus raphe magnus (NRM) of RVM in IBS model rats compared with the normal rats (P<0.05). After EA treatment, PTP values and AWR scores significantly increased and decreased, respectively (P<0.01, P<0.05); the number of immunoreactive neurons of c-fos and NR1 significantly decreased respectively in Gi, LPGi and GiA and in Gi, LPGi, GiA and NRM (P<0.05). No such effects on PTP values, AWR scores and the number of immunoreactive neurons of c-fos and NR1 were observed after sham-EA treatment.
Conclusion: These data provide the evidence that EA can relieve chronic visceral hyperalgesia in rats with IBS, and such an effect may be correlated with inhibitory modulation of hyperactivity of neurons by means of down-regulating the high expression of NR1 in RVM of IBS model rats.

Key words: receptors, N-methyl-D-aspartate, hyperalgesia, acupuncture therapy, proto-oncogene protein c-fos, irritable bowel syndrome, rats

Figure 1

RVM in the brainstem sliced at the rostral end of the Ⅳ ventricle RVM: rostral ventromedia medulla; Gi: nucleus reticularis gigantocellularis; NRM: nucleus raphe magnus; GiA: nucleus reticularis gigantocellularis pars alpha; LPGi: nucleus lateralis paragigantocellulari; 4V: the Ⅳ ventricle; Sol: solitary tract; Sp5: spinal trigeminal tract; Py: pyramidal tract; RPa: raphe pallidus nucleus; ml: medial lemniscus; 7n: facial nucleus."

Figure 2

PTP values measured in response to continuous colorectal distention Data are represented as mean±standard error of mean, n=8; **P<0.01, vs normal control group; △△P<0.01, vs IBS model control group. PTP: pain threshold pressure; IBS: irritable bowel syndrome; EA: electroacupuncture."


Group n AWR score
20 mmHg 40 mmHg 60 mmHg 80 mmHg
Normal control 8 0.30±0.09 1.39±0.12 2.26±0.14 3.02±0.22
IBS model control 8 1.24±0.18** 2.17±0.17** 3.04±0.11** 3.78±0.08**


Group n AWR score
20 mmHg 40 mmHg 60 mmHg 80 mmHg
EA treatment
Before 8 1.27±0.21 2.21±0.19 2.98±0.11 3.77±0.10
After 8 0.33±0.09▲□ 1.36±0.10▲□ 2.25±0.11▲□ 2.88±0.10▲□
Sham-EA treatment
Before 8 1.29±0.08 2.27±0.15 2.81±0.09 3.75±0.09
After 8 0.69±0.10 1.87±0.11 2.48±0.13 3.15±0.21

Figure 3

Samples of microphotographs with the c-fos- (A-D) and NR1- (E-H) immunoreactive neurons of the RVM in different groups (Light microscopy, ×25) The images of immunohistochemical results were taken under the light microscopy. The arrows show c-fos- or NR1-immunoreactive neurons. N: Normal control group; M: IBS model control group; EA: IBS model rats after EA treament; SEA: IBS model rats after sham-EA treatment; RVM: rostral ventromedia medulla; NR1: N-methyl-D-aspartate receptor 1; IBS: irritable bowel syndrome; EA: electroacupuncture."


Group n c-fos
Normal control 8 34.62±2.02 29.13±1.25 9.52±0.80 3.23±0.45
IBS model control 8 59.66±3.00* 37.34±1.95* 13.50±1.53* 4.84±0.76*
EA treatment 8 40.84±2.28 30.18±1.51 9.12±1.20 2.68±0.56
Sham-EA treatment 8 50.70±2.52 35.76±1.98 13.86±1.33 2.46±0.34


Group n NR1
Normal control 8 23.05±1.75 14.28±1.28 4.85±0.75 1.73±0.31
IBS model control 8 36.44±1.99* 21.15±1.45* 8.88±0.98* 3.00±0.50*
EA treatment 8 24.03±1.38 13.93±1.20 3.23±0.61 1.08±0.28
Sham-EA treatment 8 36.45±1.40 24.20±1.50 8.10±0.83 3.08±0.48
[1] Verne GN, Robinson ME, Price DD . Hypersensitivity to visceral and cutaneous pain in the irritable bowel syndrome. Pain. 2001; 93(1):7-14.
doi: 10.1016/S0304-3959(01)00285-8
[2] Sengupta JN . Visceral pain: the neurophysiological mechanism. Handb Exp Pharmacol. 2009; ( 194) : 31-74.
[3] Hungin AP, Whorwell PJ, Tack J, Mearin F . The prevalence, patterns and impact of irritable bowel syndrome: an international survey of 40,000 subjects. Aliment Pharmacol Ther. 2003 ; 17(5) : 643-650.
doi: 10.1046/j.1365-2036.2003.01456.x
[4] Gwee KA, Lu CL, Ghoshal UC . Epidemiology of irritable bowel syndrome in Asia: something old, something new, something borrowed.J Gastroenterol Hepatol. 2009; 24(10) : 1601-1607.
doi: 10.1111/jgh.2009.24.issue-10
[5] Cash B, Sullivan S, Barghout V . Total costs of IBS: employer and managed care perspective. Am J Manag Care. 2005; 11(1 Suppl) : S7-S16.
[6] Talley NJ, Gabriel SE, Harmsen WS, Zinsmeister AR, Evans RW . Medical costs in community subjects with irritable bowel syndrome. Gastroenterology. 1995; 109(6):1736-1741.
doi: 10.1016/0016-5085(95)90738-6
[7] Schneider A, Weiland C, Enck P, Joos S, Streitberger K, Maser-Gluth C, Zipfel S, Bagheri S, Herzog W, Friederich HC . Neuroendocrinological effects of acupuncture treatment in patients with irritable bowel syndrome. Complement Ther Med. 2007; 15(4):255-263.
doi: 10.1016/j.ctim.2006.12.002
[8] Xiao WB, Liu YL . Rectal hypersensitivity reduced by acupoint TENS in patients with diarrhea-predominant irritable bowel syndrome: a pilot study. Dig Dis Sci. 2004; 49(2) : 312-319.
doi: 10.1023/B:DDAS.0000017458.55517.33
[9] Musial F, Tao I, Dobos G . Is the analgesic effect of acupuncture a placebo effect? Schmerz. 2009 ; 23(4):341-346.
doi: 10.1007/s00482-009-0810-9
[10] Rohrbock RB, Hammer J, Vogelsang H, Talley NJ, Hammer HF . Acupuncture has a placebo effect on rectal perception but not on distensibility and spatial summation: a study in health and IBS. Am J Gastroenterol. 2004; 99(10):1990-1997.
doi: 10.1111/ajg.2004.99.issue-10
[11] Zhuo M, Gebhart GF . Facilitation and attenuation of a visceral nociceptive reflex from the rostroventral medulla in the rat. Gastroenterology. 2002 ; 122(4):1007-1019.
doi: 10.1053/gast.2002.32389
[12] Zhuo M, Sengupta JN, Gebhart GF . Biphasic modulation of spinal visceral nociceptive transmission from the rostroventral medial medulla in the rat.J Neurophysiol. 2002; 87(5) : 2225-2236.
doi: 10.1152/jn.2002.87.5.2225
[13] Urban MO, Coutinho SV, Gebhart GF . Biphasic modulation of visceral nociception by neurotensin in rat rostral ventromedial medulla.J Pharmacol Exp Ther. 1999 ; 290(1):207-213.
[14] Coutinho SV, Urban MO, Gebhart GF . Role of glutamate receptors and nitric oxide in the rostral ventromedial medulla in visceral hyperalgesia. Pain. 1998; 78(1):59-69.
doi: 10.1016/S0304-3959(98)00137-7
[15] Coutinho SV, Urban MO, Gebhart GF . The role of CNS NMDA receptors and nitric oxide in visceral hyperalgesia. Eur J Pharmacol. 2001 ; 429(1-3) : 319-325.
doi: 10.1016/S0014-2999(01)01331-0
[16] Porreca F, Ossipov MH, Gebhart GF . Chronic pain and medullary descending facilitation. Trends Neurosci. 2002; 25(6) : 319-325.
doi: 10.1016/S0166-2236(02)02157-4
[17] Al-Chaer ED, Kawasaki M, Pasricha PJ . A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development. Gastroenterology. 2000; 119(5):1276-1285.
doi: 10.1053/gast.2000.19576
[18] Cui KM, Li WM, Gao X, Chung K, Chung JM, Wu GC . Electro-acupuncture relieves chronic visceral hyperalgesia in rats. Neurosci Lett. 2005; 376(1):20-23.
doi: 10.1016/j.neulet.2004.11.018
[19] Zhou J Li WM. Influence of electroacupuncture on expression of N-methyl-D-aspartatic receptors in the spinal cord of irritable bowel syndrome rat. Shanghai Zhen Jiu Za Zhi. 2008; 27(6): 38-40. Chinese with abstract in English.
周娟, 李为民 . 电针对肠易激综合征大鼠脊髓NMDA 受体表达的影响.上海针灸杂志. 2008; 27(6):38-40.
[20] Sagar SM, Sharp FR, Curran T . Expression of c-fos protein in brain: metabolic mapping at the cellular level. Science. 1988; 240(4857):1328-1331.
doi: 10.1126/science.3131879
[21] Paxinos G, Watson C. The rat brain in stereotaxic coordinates. 6th ed. Waltham: Academic Press. 2007 : 270-298.
[22] Drossman DA . Chronic functional abdominal pain. Am J Gastroenterol. 1996; 91(11):2270-2281.
[23] Winston J, Shenoy M, Medley D, Naniwadekar A, Pasricha PJ . The vanilloid receptor initiates and maintains colonic hypersensitivity induced by neonatal colon irritation in rats. Gastroenterology. 2007; 132(2):615-627.
doi: 10.1053/j.gastro.2006.11.014
[24] Chung EK, Bian ZX, Xu HX, Sung JJ . Neonatal maternal separation increases brain-derived neurotrophic factor and tyrosine kinase receptor B expression in the descending pain modulatory system. Neurosignals. 2009; 17(3) : 213-221.
doi: 10.1159/000224631
[1] Ralph Fernando Rosas, Aline Armiliato Emer, Ana Paula Batisti, Daniela Dero Ludtke, Bruna Lenfers Turnes, Franciane Bobinski, Francisco José Cidral-Filho, Daniel Fernandes Martins. Far infrared-emitting ceramics decrease Freund’s adjuvant-induced inflammatory hyperalgesia in mice through cytokine modulation and activation of peripheral inhibitory neuroreceptors. Journal of Integrative Medicine, 2018, 16(6): 396-403.
[2] Morufu Eyitayo Balogun, Elizabeth Enohnyaket Besong, Jacinta Nkechi Obimma, Ogochukwu Sophia Mbamalu, Fankou Serges Athanase Djobissie. Protective roles of Vigna subterranea (Bambara nut) in rats with aspirin-induced gastric mucosal injury. Journal of Integrative Medicine, 2018, 16(5): 342-349.
[3] Lucky Legbosi Nwidu, Raphael Ellis Teme. Hot aqueous leaf extract of Lasianthera africana (Icacinaceae) attenuates rifampicin-isoniazid-induced hepatotoxicity. Journal of Integrative Medicine, 2018, 16(4): 263-272.
[4] Maria T.Chao, AlexandraChang, SanjayReddy, James D.Harrison, JosephAcquah, MiriaToveg, TrilceSantana, Frederick M.Hecht. Adjunctive acupuncture for pain and symptom management in the inpatient setting: Protocol for a pilot hybrid effectiveness-implementation study. Journal of Integrative Medicine, 2016, 14(3): 228-238.
[5] Paulo AraujoDias, André Brito Bastos Guimarães, Andrea de Oliveira Albuquerque, Karoline Lucasde Oliveira, Maria Luzete CostaCavalcante, Sergio BotelhoGuimarães. Short-term complementary and alternative medicine on quality of life in women with fibromyalgia. Journal of Integrative Medicine, 2016, 14(1): 29-35.
[6] Bosch Peggy, van den Noort Maurits, Yeo Sujung, Lim Sabina, Coenen Anton, van Luijtelaar Gilles. The effect of acupuncture on mood and working memory in patients with depression and schizophrenia. Journal of Integrative Medicine, 2015, 13(6): 380-390.
[7] Sung Min Lim, Sook-Hyun Lee. Effectiveness of bee venom acupuncture in alleviating post-stroke shoulder pain: A systematic review and meta-analysis. Journal of Integrative Medicine, 2015, 13(4): 241-247.
[8] Kylie Connolly, Douglas Jackson, Candice Pullen, Andrew Fenning. Alpha-adrenoceptor antagonism by Crassostrea gigas oyster extract inhibits noradrenaline-induced vascular contraction in Wistar rats. Journal of Integrative Medicine, 2015, 13(3): 194-200.
[9] Hussein O. B. Oloyede, Matthew C. Adaja, Taofeek O. Ajiboye, Musa O. Salawu. Anti-ulcerogenic activity of aqueous extract of Carica papaya seed on indomethacin-induced peptic ulcer in male albino rats. Journal of Integrative Medicine, 2015, 13(2): 105-114.
[10] Udhaya Lavinya Baskaran, Sherry Joseph Martin, Rasool Mahaboobkhan, Sabina Evan Prince. Protective role of Triphala, an Indian traditional herbal formulation, against the nephrotoxic effects of bromobenzene in Wistar albino rats. Journal of Integrative Medicine, 2015, 13(2): 115-121.
[11] Marco Gemma, Elisa Nicelli, Luigi Gioia, Elena Moizo, Luigi Beretta, Maria Rosa Calvi. Acupuncture accelerates recovery after general anesthesia: A prospective randomized controlled trial. Journal of Integrative Medicine, 2015, 13(2): 99-104.
[12] Anirudha A. Lande, Shirishkumar D. Ambavade, Uma S. Swami, Prafulla P. Adkara Prashant D. Ambavade, Arun B. Waghamare. Saponins isolated from roots of Chlorophytum borivilianum reduce acute and chronic inflammation and histone deacetylase. Journal of Integrative Medicine, 2015, 13(1): 25-33.
[13] Kwan Leung Chia. Symptomatic Tarlov cyst and electroacupuncture: More studies required. Journal of Integrative Medicine, 2015, 13(1): 58-60.
[14] Elizabeth Abidemi Balogun, Sylvia Orume Malomo, Joseph Oluwatope Adebayo, Ahmed Adebayo Ishola, Ayodele Olufemi Soladoye, Lawrence Aderemi Olatunji, Olatunji Matthew Kolawole, Stephen Olubunmi Oguntoye, Abiola Samuel Babatunde, Oluwole Busayo Akinola . In vivo antimalarial activity and toxicological effects of methanolic extract of Cocos nucifera (Dwarf red variety) husk fibre. Journal of Integrative Medicine, 2014, 12(6): 504-511.
[15] Xin-fang Zhang, Ji Zhu, Wen-ye Geng, Shu-jun Zhao, Chuan-wei Jiang, Sheng-rong Cai, Miao Cheng, Chuan-yun Zhou, Zi-bing Liu. Electroacupuncture at Feishu (BL13) and Zusanli (ST36) down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease. Journal of Integrative Medicine, 2014, 12(5): 417-424.
Full text



[1] Wei-xiong Liang. Problems-solving strategies in clinical treatment guideline for traditional Chinese medicine and integrative medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 1-4
[2] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. An explanation on "guiding principles of clinical research on mild cognitive impairment (protocol)". Journal of Chinese Integrative Medicine, 2008, 6(1): 15-21
[3] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[4] Yi-ting He, Qing-lin Zha, Jian-ping Yu, Yong Tan, Cheng Lu, Ai-ping Lv. Principal factor analysis of symptoms of rheumatoid arthritis and their correlations with efficacy of traditional Chinese medicine and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 32-36
[5] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[6] Wei Zhang, Xiang-feng Lu, Xiao-mei Zhang, Jian-jun Wu, Liang-duo Jiang. A rat model of pulmonary fibrosis induced by infusing bleomycin quickly through tracheal intubation. Journal of Chinese Integrative Medicine, 2008, 6(1): 60-67
[7] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[8] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[9] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. Guiding principles of clinical research on mild cognitive impairment (protocol). Journal of Chinese Integrative Medicine, 2008, 6(1): 9-14
[10] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97