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

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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
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