title: The TRPV1 channel in rodents is a major target for antinociceptive effect of the probiotic Lactobacillus reuteri DSM 17938.
abstract: Certain probiotic bacteria have been shown to reduce distension-dependent gut pain, but the mechanisms involved remain obscure. Live luminal Lactobacillus reuteri (DSM 17938) and its conditioned medium dose dependently reduced jejunal spinal nerve firing evoked by distension or capsaicin, and 80% of this response was blocked by a specific TRPV1 channel antagonist or in TRPV1 knockout mice. The specificity of DSM action on TRPV1 was further confirmed by its inhibition of capsaicin-induced intracellular calcium increases in dorsal root ganglion neurons. Another lactobacillus with ability to reduce gut pain did not modify this response. Prior feeding of rats with DSM inhibited the bradycardia induced by painful gastric distension. These results offer a system for the screening of new and improved candidate bacteria that may be useful as novel therapeutic adjuncts in gut pain. Certain bacteria exert visceral antinociceptive activity, but the mechanisms involved are not determined. Lactobacillus reuteri DSM 17938 was examined since it may be antinociceptive in children. Since transient receptor potential vanilloid 1 (TRPV1) channel activity may mediate nociceptive signals, we hypothesized that TRPV1 current is inhibited by DSM. We tested this by examining the effect of DSM on the firing frequency of spinal nerve fibres in murine jejunal mesenteric nerve bundles following serosal application of capsaicin. We also measured the effects of DSM on capsaicin-evoked increase in intracellular Ca(2+) or ionic current in dorsal root ganglion (DRG) neurons. Furthermore, we tested the in vivo antinociceptive effects of oral DSM on gastric distension in rats. Live DSM reduced the response of capsaicin- and distension-evoked firing of spinal nerve action potentials (238 ± 27.5% vs. 129 ± 17%). DSM also reduced the capsaicin-evoked TRPV1 ionic current in DRG neuronal primary culture from 83 ± 11% to 41 ± 8% of the initial response to capsaicin only. Another lactobacillus (Lactobacillus rhamnosus JB-1) with known visceral anti-nociceptive activity did not have these effects. DSM also inhibited capsaicin-evoked Ca(2+) increase in DRG neurons; an increase in Ca(2+) fluorescence intensity ratio of 2.36 ± 0.31 evoked by capsaicin was reduced to 1.25 ± 0.04. DSM releasable products (conditioned medium) mimicked DSM inhibition of capsaicin-evoked excitability. The TRPV1 antagonist 6-iodonordihydrocapsaicin or the use of TRPV1 knock-out mice revealed that TRPV1 channels mediate about 80% of the inhibitory effect of DSM on mesenteric nerve response to high intensity gut distension. Finally, feeding with DSM inhibited perception in rats of painful gastric distension. Our results identify a specific target channel for a probiotic with potential therapeutic properties.
authors.affiliation: McMaster Brain-Body Institute, St Joseph's Healthcare, Hamilton, 50 Charlton Avenue East, Hamilton, Ontario, Canada, L8N 4A6.
authors.forename: Azucena
authors.initials: A
authors.lastname: Perez-Burgos
authors.affiliation: Farncombe Family Digestive Health Research Institute, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.affiliation: McMaster Brain-Body Institute, St Joseph's Healthcare, Hamilton, 50 Charlton Avenue East, Hamilton, Ontario, Canada, L8N 4A6.
authors.forename: Karen-Anne
authors.initials: KA
authors.lastname: McVey Neufeld
authors.affiliation: McMaster Brain-Body Institute, St Joseph's Healthcare, Hamilton, 50 Charlton Avenue East, Hamilton, Ontario, Canada, L8N 4A6.
authors.forename: Yu-Kang
authors.initials: YK
authors.lastname: Mao
authors.affiliation: Firestone Institute for Respiratory Health, St Joseph's Hospital, Department of Medicine, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Ahmadzai
authors.affiliation: Firestone Institute for Respiratory Health, St Joseph's Hospital, Department of Medicine, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Luke J
authors.initials: LJ
authors.lastname: Janssen
authors.affiliation: McMaster Brain-Body Institute, St Joseph's Healthcare, Hamilton, 50 Charlton Avenue East, Hamilton, Ontario, Canada, L8N 4A6.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Stanisz
authors.affiliation: McMaster Brain-Body Institute, St Joseph's Healthcare, Hamilton, 50 Charlton Avenue East, Hamilton, Ontario, Canada, L8N 4A6.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada.
authors.forename: John
authors.initials: J
authors.lastname: Bienenstock
authors.affiliation: McMaster Brain-Body Institute, St Joseph's Healthcare, Hamilton, 50 Charlton Avenue East, Hamilton, Ontario, Canada, L8N 4A6.
authors.forename: Wolfgang A
authors.initials: WA
authors.lastname: Kunze
mesh_terms: D000698:Analgesia; D000818:Animals; D001919:Bradycardia / Q000209:etiology / Q000503:physiopathology / Q000628:therapy*; D002211:Capsaicin; D005727:Ganglia, Spinal / Q000502:physiology; D007583:Jejunum / Q000294:innervation / Q000502:physiology*; D052200:Limosilactobacillus reuteri; D008297:Male; D008643:Mesentery / Q000294:innervation / Q000502:physiology; D018345:Mice, Knockout; D019936:Probiotics / Q000494:pharmacology / Q000627:therapeutic use; D017207:Rats, Sprague-Dawley; D013127:Spinal Nerves / Q000502:physiology; D013272:Stomach Diseases / Q000150:complications / Q000503:physiopathology / Q000628:therapy*; D050916:TRPV Cation Channels / Q000235:genetics / Q000502:physiology*
chemical_list: D050916:TRPV Cation Channels; C494484:TRPV1 receptor; D002211:Capsaicin