Inhibitory Effects of Botulinum Toxin Type A on Pyloric Cholinergic Muscle Contractility of Rat

10.4077/CJP.2016.BAE401

Peng Zhao；Hong-Xu Sun；Min Chu；Yi-Ping Hou

acetylcholine ； botulinum toxin type A ； pyloric smooth muscle ； synaptosomal-associated protein of 25 kDa ； toosendanin

The Chinese Journal of Physiology

59卷4期（2016 / 08 / 31）

218 - 224

英文

Botulinum toxin type A (BTX-A) selectively cleaves synaptosomal-associated protein of 25 kDa (SNAP-25) and results in inhibition of the fusion of synaptic vesicles containing neurotransmitters with the presynaptic membrane to undergo exocytosis and release. The aim of this study was to investigate whether BTX-A inhibited the pyloric smooth muscle contractility induced by acetylcholine (ACh) after BTX-A-mediated cleavage of SNAP-25 antagonized by toosendanin (TSN). Three groups of rat pyloric muscle strips were studied in vitro. All strips were allowed to equilibrate for 52 min under a basal loading tension of 1 g in Krebs solution and spontaneous contractile waves were recorded as their own controls before adding each drug. According to experimental protocols, 100 μM ACh, 1 μM atropine, 29.6 μM TSN and 10 U/ml BTX-A was added, respectively. BTX-A directly inhibited pyloric spontaneous contraction and ACh-induced contractile response. Addition of 10 U/ml BTX-A still inhibited pyloric smooth muscle contractility following incubation of TSN, while subsequent administration of 100 μM ACh had no effect. BTX-A inhibits pyloric smooth muscle contractility in our study suggesting BTX-A inhibits not only ACh release from cholinergic nerves but also muscarinic cholinergic muscular transmission.

1. Lin, C.C.,Chang, C.W.,Tsai, S.J.(2015).Optimal effect of phenol block in the sciatic nerve of rat: standardization of minimized dosage and duration of application.Chinese J. Physiol.,58,237-243.
   連結：
2. Blasi, J.,Chapman, E.R.,Link, E.,Binz, T.,Yamasaki, S.,De Camilli, P.,Südhof, T.C.,Niemann, H.,Jahn, R.(1993).Botulinum toxin A selectively cleaves the synaptic protein SNAP-25.Nature,365,160-163.
3. Brin, M.F.,Fahn, S.,Moskowitz, C.,Friedman, A.,Shale, H.M.,Greene, P.E.,Blitzer, A.,List, T.,Lange, D.,Lovelace, R.E.,McMahon, M.(1987).Localized injections of botulinum toxin for the treatment of focal dystonia and hemifacial spasm.Mov. Disord.,2,237-254.
4. Castro, W.H.,Gomez, R.S.,da Silva Oliveira, J.,Moura, M.D.,Gomez, R.S.(2005).Botulinum toxin type A in the management of masseter muscle hypertrophy.J. Oral Maxillofac. Surg.,63,20-24.
5. Dolly, J.O.(2005).Molecular definition of neuronal targets for novel neurotherapeutics: SNAREs and Kv1 channels.Neurotoxicology,26,753-760.
6. Dressler, D.,Saberi, F.A.,Barbosa, E.R.(2005).Botulinum toxinmechanisms of action.Arq. Neuropsiquiatr.,63,180-185.
7. Eswaramoorthy, S.,Kumaran, D.,Swaminathan, S.(2002).A novel mechanism for clostridium botulinum neurotoxin inhibition.Biochemistry,41,9795-9802.
8. Ezzeddine, D.,Jit, R.,Katz, N.,Gopalswamy, N.,Bhutani, M.S.(2002).Pyloric injection of botulinum toxin for treatment of diabetic gastroparesis.Gastrointest. Endosc.,55,920-923.
9. Goonetilleke, A.,Harris, J.B.(2004).Clostridal neurotoxins.J. Neurol. Neurosurg. Psychiatry,75(Suppl 3),iii35- iii39.
10. Guarda-Nardini, L.,Manfredini, D.,Salamone, M.,Salmaso, L.,Tonello, S.,Ferronato, G.(2008).Efficacy of botulinum toxin in treating myofascial pain in bruxers: a controlled placebo pilot study.Cranio,26,126-135.
11. Hou, Y.P.,Zhang, Y.P.,Song, Y.F.,Zhou, C.M.,Wang, Y.C.,Xie, G.L.(2007).Botulinum toxin type A inhibits rat pyloric myoelectrical activityand substance P release in vivo.Can. J. Physiol. Pharmacol.,85,209-214.
12. Jahn, R.,Lang, T.,Sudhof, T.C.(2003).Membrane fusion.Cell,112,519-533.
13. James, A.N.,Ryan, J.P.,Parkman, H.P.(2003).Inhibitory effects of botulinum toxin on pyloric and antral smooth muscle.Am. J. Physiol. Gastrointest. Liver Physiol.,285,G291-G297.
14. Jankovic, J.,Brin, M.F.(1991).Therapeutic uses of botulinum toxin.N. Engl. J. Med.,324,1186-1194.
15. Ji, J.,Salapatek, A.M.F.,Lau, H.,Wang, G.,Gaisano, H.Y.,Diamant, H.E.(2002).SNAP-25, a SNARE protein, inhibits two types of K+ channels in esophageal smooth muscle.Gastroenterology,122,994-1006.
16. Lacy, B.E.,Zayat, E.N.,Crowell, M.D.,Schuster, M.M.(2002).Botulinum toxin for the treatment of gastroparesis: a preliminary report.Am. J. Gastroenterol.,97,1548-1552.
17. Lawrence, G.W.,Foran, P.,Oliver Dolly, J.(2002).Insights into a basis for incomplete inhibition by botulinum toxin A of Ca2+-evoked exocytosis from permeabilised chromaffin cells.Toxicology,181-182,249-253.
18. Lee, S.J.,McCall, W.D., Jr.,Kim, Y.K.,Chung, S.C.,Chung, J.W.(2010).Effect of botulinum toxin injection on nocturnal bruxism: a randomized controlled trial.Am. J. Phys. Med. Rehabil.,89,16-23.
19. Li, C.,Micci, M.A.,Murthy, K.S.,Pasricha, P.J.(2014).Substance P is essential for maintaining gut muscle contractility: a novel role for coneurotransmission revealed by botulinum toxin.Am. J. Physiol. Gastrointest. Liver Physiol.,306,G839-G848.
20. Llewellyn-Smith, I.J.,Furness, J.B.,Gibbins, I.L.,Costa, M.(1988).Quantitative ultrastructural analysis of enkephalin-, substance p-, and vip-immunoreactive nerve fibers in the circular muscle of the guinea pig small intestine.J. Comp. Neurol.,272,139-148.
21. Ma, J.,Shen, J.,Lee, C.A.,Elsaidi, G.A.,Smith, T.L.,Walker, F.O.,Rushing, J.T.,Tan, K.H.,Koman, L.A.,Smith, B.P.(2005).Gene expression of nAChR, SNAP-25 and GAP-43 in skeletal muscles following botulinum toxin A injection: a study in rats.J. Orthop. Res.,23,302-309.
22. Mackenzie, I.,Burnstock, G.,Dolly, J.O.(1982).The effects of purified botulinum neurotoxin type A on cholinergic, and adrenergic and non-adrenergic, atropine-resistant autonomic neuromuscular transmission.Neuroscience,7,997-1006.
23. Mearin, F.,Camilleri, M.,Malagelada, J.R.(1986).Pyloric dysfunction in diabetics with recurrent nausea and vomiting.Gastroenterology,90,1919-1925.
24. Mense, S.(2004).Neurobiological basis for the use of botulinum toxin in pain therapy.J. Neurol.,251(Suppl 1),I1- I7.
25. Miller, L.S.,Szych, G.A.,Kantor, S.B.,Bromer, M.Q.,Knight, L.C.,Mauer, A.H.,Fisher, R.S.,Parkman, H.P.(2002).Treatment of idiopathic gastroparesis with injection of botulinum toxin into the pyloric sphincter muscle.Am. J. Gastroenterol.,97,1653-1660.
26. Persaud, R.,Garas, G.,Silva, S.,Stamatoglou, C.,Chatrath, P.,Patel, K.(2013).An evidence-based review of botulinum toxin (Botox) applications in non-cosmetic head and neck conditions.JRSM Short Rep.,4,10.
27. Rao, K.N.,Kumaran, D.,Binz, T.,Swaminathan, S.(2005).Structural analysis of the catalytic domain of tetanus neurotoxin.Toxicon,45,929-939.
28. Reddymasu, S.C.,Singh, S.,Sankula, R.,Lavenbarg, T.A.,Olyaee, M.,McCallum, R.W.(2009).Endoscopic pyloric injection of botulinum toxin-A for the treatment of postvagotomy gastroparesis.Am. J. Med. Sci.,337,161-164.
29. Rossetto, O.,Rigoni, M.,Montecucco, C.(2004).Different mechanism of blockade of neuroexocytosis by presynaptic neurotoxins.Toxicol. Lett.,149,91-101.
30. Saffrey, M.J.(2004).Ageing of the enteric nervous system.Mech. Ageing Dev.,125,899-906.
31. Sand, J.,Nordback, I.,Arvola, P.,Porsti, I.,Kalloo, A.,Pasricha, P.(1998).Effects of botulinum toxin A on the sphincter of Oddi: an in vivo and in vitro study.Gut,42,507-510.
32. Shao, Y.F.,Xie, J.F.,Ren, Y.X.,Wang, C.,Kong, X.P.,Zong, X.J.,Fan, L.L.,Hou, Y.P.(2015).The inhibitory effect of botulinum toxin type A on rat pyloric smooth muscle contractile response to substance P in vitro.Toxins,7,4143-4156.
33. Shi, Y.L.,Wang, Z.F.(2004).Cure of experimental botulism and antibotulismic effect of toosendanin.Acta Pharmacol. Sin.,25,839-848.
34. Simpson, L.L.(1986).Molecular pharmacology of botulinum toxin and tetanus toxin.Annu. Rev. Pharmacol. Toxicol.,26,427-453.
35. Simpson, L.L.,Maksymowych, A.B.,Kouguchi, H.,Dubois, G.,Bora, R.S.,Joshi, S.(2005).The role of exoproteases in governing intraneuronal metabolism of botulinum toxin.Protein J.,24,155-165.
36. Steele, P.A.,Brookes, S.J.,Costa, M.(1991).Immunohistochemical identification of cholinergic neurons in the myenteric plexus of guinea-pig small intestine.Neuroscience,45,227-239.
37. Taylor, G.S.,Bywater, R.A.(1986).Antagonism of non-cholinergic excitatory junction potentials in the guinea-pig ileum by a substance p analogue antagonist.Neurosci. Lett.,63,23-26.
38. Washbourne, P.,Pellizzari, R.,Baldini, G.,Wilson, M.C.,Montecucco, C.(1997).Botulinum neurotoxin types A and E require the SNARE motif in SNAP-25 for proteolysis.FEBS Lett.,418,1-5.
39. Zhou, J.Y.,Wang, Z.F.,Ren, X.M.,Tang, M.Z.,Shi, Y.L.(2003).Antagonism of botulinum toxin type A-induced cleavage of SNAP-25 in rat cerebral synaptosome by toosendanin.FEBS Lett.,555,375-379.