Korean J Physiol Pharmacol.  2016 Jul;20(4):407-414. 10.4196/kjpp.2016.20.4.407.

Involvement of spinal muscarinic and serotonergic receptors in the anti-allodynic effect of electroacupuncture in rats with oxaliplatin-induced neuropathic pain

• Lee JH
• Go D
• Kim W
• Lee G
• Bae H
• Quan FS
• Kim SK

• KMID: 2371058
• DOI: http://doi.org/10.4196/kjpp.2016.20.4.407

Abstract

This study was performed to investigate whether the spinal cholinergic and serotonergic analgesic systems mediate the relieving effect of electroacupuncture (EA) on oxaliplatin-induced neuropathic cold allodynia in rats. The cold allodynia induced by an oxaliplatin injection (6 mg/kg, i.p.) was evaluated by immersing the rat's tail into cold water (4℃) and measuring the withdrawal latency. EA stimulation (2 Hz, 0.3-ms pulse duration, 0.2~0.3 mA) at the acupoint ST36, GV3, or LI11 all showed a significant anti-allodynic effect, which was stronger at ST36. The analgesic effect of EA at ST36 was blocked by intraperitoneal injection of muscarinic acetylcholine receptor antagonist (atropine, 1 mg/kg), but not by nicotinic (mecamylamine, 2 mg/kg) receptor antagonist. Furthermore, intrathecal administration of M(2) (methoctramine, 10 µg) and M(3) (4-DAMP, 10 µg) receptor antagonist, but not M(1) (pirenzepine, 10 µg) receptor antagonist, blocked the effect. Also, spinal administration of 5-HT(3) (MDL-72222, 12 µg) receptor antagonist, but not 5-HT(1A) (NAN-190, 15 µg) or 5-HT(2A) (ketanserin, 30 µg) receptor antagonist, prevented the anti-allodynic effect of EA. These results suggest that EA may have a signifi cant analgesic action against oxaliplatin-induced neuropathic pain, which is mediated by spinal cholinergic (M(2), M(3)) and serotonergic (5-HT(3)) receptors.

Keyword

Acetylcholine; Cold allodynia; Electroacupuncture; Oxaliplatin; Serotonin

MeSH Terms

Acetylcholine
Acupuncture Points
Animals
Electroacupuncture*
Hyperalgesia
Injections, Intraperitoneal
Neuralgia*
Rats*
Receptors, Muscarinic
Serotonin
Tail
Water
Acetylcholine
Receptors, Muscarinic
Serotonin
Water

Figure

• 

  Fig. 1 Experimental schedule and acupoint specific effect of EA on oxaliplatin-induced cold allodynia.(A) Behavioral tests for cold allodynia were performed before and 20 min after EA treatment in the three groups: (B) LI11 (n=8), (C) GV3 (n=6), and (D) ST36 (n=10). The effect of EA at LI11, GV3 and ST36 significantly increased the TWL. (E) Potency of EA against cold allodynia was significantly greater at GV3 and ST36 comparing with LI11. Data are presented as mean±S.E.M.; ***p<0.001, **p<0.01, *p<0.05; by paired t-test.

• 

  Fig. 2 Effects of non-selective muscarinic and nicotinic acetylcholine receptor antagonists on EA-induced anti-allodynia.Rats with cold allodynia induced by a single injection of oxaliplatin were randomly divided into three groups. The behavioral tests for cold allodynia were performed before an intraperitoneal injection of antagonists and after EA treatment: (A) NS (n=5), (B) atropine (n=5), and (C) mecamylamine (n=7). The analgesic effect of EA at ST36 was blocked by intraperitoneal injection of atropine (non-selective muscarinic receptor antagonist, 1 mg/kg), but not by intraperitoneal injection of mecamylamine (non-selective nicotinic receptor antagonist, 2 mg/kg). Data are presented as mean±S.E.M.; **p<0.01; by paired t-test.

• 

  Fig. 3 Effects of spinal muscarinic acetylcholine receptor subtype antagonists on EA-induced anti-allodynia.The behavioral tests for cold allodynia were performed before the intrathecal injection of receptor antagonists and after EA treatment in four groups: (A) NS (n=4), (B) pirenzepine (n=6), (C) methoctramine (n=8), and (D) 4-DAMP (n=7). Methoctramine (M2 receptor antagonist), and 4-DAMP (M3 receptor antagonist) blocked the analgesic effect of EA, whereas, NS and pirenzepine (M1 receptor antagonist) did not block the analgesic effect of EA. Data are presented as mean±S.E.M.; **p<0.01; by paired t-test.

• 

  Fig. 4 Effect of spinal serotonergic receptor subtypes antagonist on EA-induced anti-allodynia.The behavioral tests for cold allodynia were performed before the intrathecal injection of antagonists and after EA treatment in four groups: (A) DMSO (n=4), (B) NAN-190 (n=6), (C) ketanserin (n=8), and (D) MDL-72222 (n=6). MDL-72222 (5-HT3 receptor antagonist) completely blocked the analgesic effect of EA; however, DMSO, NAN-190 (5-HT1A receptor antagonist), and ketanserin (5-HT2A receptor antagonist) did not alter the anti-allodynic effect of EA. Data are presented as mean±SEM.; ***p<0.001, **p<0.01, *p<0.05; by paired t-test.

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