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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (12): 1490-1496.doi: 10.3736/jcim20121224

• Original Experimental Research • Previous Articles     Next Articles

Effects of electroacupuncture on expression of c-fos protein in the spinal dorsal horn of rats with chronic visceral hyperalgesia

De-bo Qi, Wei-min Li()   

  1. Laboratory of Neuronal Network and Systems Biology, Shanghai Medical College, Fudan University, Shanghai 200032, China
  • Received:2012-06-10 Accepted:2012-07-04 Online:2012-12-20 Published:2018-12-15

Objective: Acupuncture is widely used in clinics to suppress chronic visceral pain in patients with irritable bowel syndrome (IBS); however, the exact neurobiological mechanisms for its therapeutic effects need further exploration. The aim of this study was to investigate the possible involvement of spinal neurons in the effects of electroacupuncture (EA) in relieving chronic visceral hyperalgesia in a rat model of IBS.
Methods: Colon mechanical irritation was applied to male neonatal Sprague-Dawley rats to establish the IBS model. Behavioral test of the abdominal withdraw reflex (AWR) response to colorectal distention stimuli was conducted to judge the degree of colorectal sensitivity. EA at acupoints Zusanli (ST36) and Shangjuxu (ST37) was applied bilaterally in a total of four times every other day, while sham-EA at similar acupoints was done by inserting needles without electrical stimulation. Immunohistochemical methods were used to display the expression of proto-oncogene protein c-fos in the spinal dorsal horn.
Results: It was found that AWR scores were significantly increased in the IBS model rats (P<0.01), accompanied with significant increase in the expression of c-fos protein in the superficial laminae (SDH, laminae Ⅰ and Ⅱ) and nucleus proprius (NP, laminae Ⅲ and Ⅳ), the neck of the dorsal horn (NECK, laminae V and VI) at lumbosacral (L6-S2) spinal level, and in NECK at thoracolumbar (T13-L2) spinal level, when compared with normal rats (P<0.05). After EA treatment, AWR scores and the expression of c-fos protein in SDH, NP and NECK at similar spinal levels were significantly decreased in the IBS model rats (P<0.05). No such effects on either AWR scores or the expression of c-fos protein were observed in IBS model rats after sham-EA treatment.
Conclusion: The abnormally high neuronal excitability in the spinal dorsal horn may be an important reason underlying the visceral hyperalgesia in IBS model rats. EA treatment can relieve the chronic visceral hyperalgesia in IBS rats by suppressing the abnormal neuronal excitability in the spinal dorsal horn.

Key words: hyperalgesia, acupuncture therapy, proto-oncogene proteins c-fos, irritable bowel syndrome, rats

Figure 1

Schematic diagram for measuring AWR scores in rats AWR responses to graded CRD of 20, 40, 60 and 80 mmHg were visually observed and were divided into 5 scores: 0, no behavioral response to CRD; 1, an immobility response during the CRD at the onset of the stimulus; 2, a mild contraction of the abdominal muscles, but no lifting the abdomen off the platform; 3, a strong contraction of the abdominal muscles and lifting the abdomen off the platform, no lifting the pelvic structure off the platform; 4, body arching and lifting the pelvic structure and scrotum. AWR: abdominal withdrawal reflex; CRD: colorectal distention."

Figure 2

The spinal dorsal horn sliced at the level of T13-L2 and L6-S2 segments A: The sample of transversal photographs at the L1 level; B: The sample of transversal photographs at the L6 level. SDH: superficial laminae (laminae Ⅰ and Ⅱ); NP: nucleus proprius (laminae Ⅲ and Ⅳ); NECK: neck of the dorsal horn (laminae Ⅴ and Ⅵ); 1-10: spinal laminae Ⅰ - Ⅹ."

"

Group n 20 mmHg 40 mmHg 60 mmHg 80 mmHg
Normal control 8 0.35±0.10 1.38±0.09 2.04±0.09 2.67±0.07
IBS model 8 1.00±0.09** 2.19±0.07** 2.90±0.08** 3.67±0.12**
EA treatment 8 0.29±0.06 1.39±0.05 2.32±0.07 3.01±0.07
Sham-EA treatment 8 0.77±0.05 1.91±0.07 2.79±0.07 3.60±0.08

Figure 3

Representative microphotographs of c-fos protein immunoreactive neurons in thoracolumbar segments (Light microscopy, ×100) N: Normal control group; M: IBS model control group; EA: IBS model rats after EA treatment; SEA: IBS model rats after sham-EA treatment; IBS: irritable bowel syndrome; EA: electroacupuncture."

"

Group n SDH NP NECK Central canal region
Normal control 8 29.67±1.30 18.94±1.39 10.16±0.95 18.73±1.25
IBS model 8 28.36±1.14 22.10±1.02 14.30± 0.80* 18.60±0.87
EA treatment 8 27.09±1.42 19.23±0.94 9.74±0.74 16.04±0.88
Sham-EA treatment 8 27.84±1.03 22.66±1.14 10.83±0.73 15.37±0.88

Figure 4

Representative microphotographs of c-fos protein immunoreactive neurons in lumbosacral segments (Light microscopy, ×100) N: Normal control group; M: IBS model control group; EA: IBS model rats after EA treatment; SEA: IBS model rats after sham-EA treatment; IBS: irritable bowel syndrome; EA: electroacupuncture."

"

Group n SDH NP NECK Central canal region
Normal control 8 19.37±0.71 19.39±0.68 18.25±0.72 9.09±0.45
IBS model 8 30.77±1.29* 35.38±1.39* 24.87±1.03* 11.36±1.0*
EA treatment 8 22.33±0.85 21.48±0.82 15.94±0.88 7.32±0.45
Sham-EA treatment 8 27.89±1.04 28.85±1.16 23.05±1.14 10.58±0.67
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