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Chemokine CXCL13 mediates orofacial neuropathic pain via CXCR5/ERK pathway in the trigeminal ganglion of mice.

Zhang Q, Cao DL, Zhang ZJ, Jiang BC, Gao YJ - J Neuroinflammation (2016)

Bottom Line: The effect of shRNA targeting on CXCL13 or CXCR5 on pain hypersensitivity was checked by behavioral testing. pIONL induced persistent mechanical allodynia and increased the expression of ATF3, CXCL13, and CXCR5 in the TG.Furthermore, MEK inhibitor (PD98059) attenuated mechanical allodynia and reduced TNF-α and IL-1β upregulation induced by pIONL.Pretreatment with PD98059, Etanercept, or Diacerein partially blocked CXCL13-induced mechanical allodynia, and PD98059 also reduced CXCL13-induced TNF-α and IL-1β upregulation.

View Article: PubMed Central - PubMed

Affiliation: Pain Research Laboratory, Institute of Nautical Medicine, Jiangsu Key Laboratory of Inflammation and Molecular Drug Target, Nantong University, Seyuan Road, Nantong, Jiangsu, 226019, China.

ABSTRACT

Background: Trigeminal nerve damage-induced neuropathic pain is a severely debilitating chronic orofacial pain syndrome. Spinal chemokine CXCL13 and its receptor CXCR5 were recently demonstrated to play a pivotal role in the pathogenesis of spinal nerve ligation-induced neuropathic pain. Whether and how CXCL13/CXCR5 in the trigeminal ganglion (TG) mediates orofacial pain are unknown.

Methods: The partial infraorbital nerve ligation (pIONL) was used to induce trigeminal neuropathic pain in mice. The expression of ATF3, CXCL13, CXCR5, and phosphorylated extracellular signal-regulated kinase (pERK) in the TG was detected by immunofluorescence staining and western blot. The effect of shRNA targeting on CXCL13 or CXCR5 on pain hypersensitivity was checked by behavioral testing.

Results: pIONL induced persistent mechanical allodynia and increased the expression of ATF3, CXCL13, and CXCR5 in the TG. Inhibition of CXCL13 or CXCR5 by shRNA lentivirus attenuated pIONL-induced mechanical allodynia. Additionally, pIONL-induced neuropathic pain and the activation of ERK in the TG were reduced in Cxcr5 (-/-) mice. Furthermore, MEK inhibitor (PD98059) attenuated mechanical allodynia and reduced TNF-α and IL-1β upregulation induced by pIONL. TNF-α inhibitor (Etanercept) and IL-1β inhibitor (Diacerein) attenuated pIONL-induced orofacial pain. Finally, intra-TG injection of CXCL13 induced mechanical allodynia, increased the activation of ERK and the production of TNF-α and IL-1β in the TG of WT mice, but not in Cxcr5 (-/-) mice. Pretreatment with PD98059, Etanercept, or Diacerein partially blocked CXCL13-induced mechanical allodynia, and PD98059 also reduced CXCL13-induced TNF-α and IL-1β upregulation.

Conclusions: CXCL13 and CXCR5 contribute to orofacial pain via ERK-mediated proinflammatory cytokines production. Targeting CXCL13/CXCR5/ERK/TNF-α and IL-1β pathway in the trigeminal ganglion may offer effective treatment for orofacial neuropathic pain.

No MeSH data available.


Related in: MedlinePlus

Inhibition of CXCL13 by shRNA lentivirus attenuated pIONL-induced mechanical allodynia. a Pretreatment with LV-Cxcl13 shRNA increased the contact time between 3 and 21 days after the operation, compared to LV-NC-injected mice. Intra-TG injection was performed 7 days before pIONL (arrow). *P < 0.05, **P < 0.01, ***P < 0.001. Two-way repeated measures ANOVA followed by Bonferroni test. b Posttreatment with LV-Cxcl13 shRNA increased the contact time between 7 and 14 days after the operation, compared to LV-NC treatment. Intra-TG injection was performed 3 days after pIONL (arrow). *P < 0.05, **P < 0.01. Two-way repeated measures ANOVA followed by Bonferroni test. c Representative fluorescence photomicrograph shows GFP expression in the TG 7 days after intra-TG infusion of lentivirus vector. d Real-time PCR assay of Cxcl13 shows that pretreatment with LV-Cxcl13 shRNA inhibited pIONL-induced Cxcl13 upregulation. **P < 0.01, ***P < 0.001. Student’s t test
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Fig3: Inhibition of CXCL13 by shRNA lentivirus attenuated pIONL-induced mechanical allodynia. a Pretreatment with LV-Cxcl13 shRNA increased the contact time between 3 and 21 days after the operation, compared to LV-NC-injected mice. Intra-TG injection was performed 7 days before pIONL (arrow). *P < 0.05, **P < 0.01, ***P < 0.001. Two-way repeated measures ANOVA followed by Bonferroni test. b Posttreatment with LV-Cxcl13 shRNA increased the contact time between 7 and 14 days after the operation, compared to LV-NC treatment. Intra-TG injection was performed 3 days after pIONL (arrow). *P < 0.05, **P < 0.01. Two-way repeated measures ANOVA followed by Bonferroni test. c Representative fluorescence photomicrograph shows GFP expression in the TG 7 days after intra-TG infusion of lentivirus vector. d Real-time PCR assay of Cxcl13 shows that pretreatment with LV-Cxcl13 shRNA inhibited pIONL-induced Cxcl13 upregulation. **P < 0.01, ***P < 0.001. Student’s t test

Mentions: To examine the role of CXCL13 in the development and maintenance of trigeminal neuropathic pain, we injected Cxcl13 shRNA lentivirus vectors (LV-Cxcl13 shRNA) into the TG before or after pIONL. Our previous study has shown that LV-Cxcl13 shRNA effectively decreased CXCL13 expression both in vitro and in vivo [11]. As shown in Fig. 3a, compared to control lentivirus injection (LV-NC), pretreatment with LV-Cxcl13 shRNA (7 days before pIONL) inhibited pIONL-induced mechanical allodynia for more than 21 days (P < 0.001, two-way repeated measures ANOVA). In addition, posttreatment with LV-Cxcl13 shRNA (3 days after pIONL) also attenuated mechanical allodynia from day 7 to day 14 (P < 0.001, two-way repeated measures ANOVA, Fig. 3b). These data suggest that CXCL13 plays an important role in both development and maintenance of trigeminal neuropathic pain.Fig. 3


Chemokine CXCL13 mediates orofacial neuropathic pain via CXCR5/ERK pathway in the trigeminal ganglion of mice.

Zhang Q, Cao DL, Zhang ZJ, Jiang BC, Gao YJ - J Neuroinflammation (2016)

Inhibition of CXCL13 by shRNA lentivirus attenuated pIONL-induced mechanical allodynia. a Pretreatment with LV-Cxcl13 shRNA increased the contact time between 3 and 21 days after the operation, compared to LV-NC-injected mice. Intra-TG injection was performed 7 days before pIONL (arrow). *P < 0.05, **P < 0.01, ***P < 0.001. Two-way repeated measures ANOVA followed by Bonferroni test. b Posttreatment with LV-Cxcl13 shRNA increased the contact time between 7 and 14 days after the operation, compared to LV-NC treatment. Intra-TG injection was performed 3 days after pIONL (arrow). *P < 0.05, **P < 0.01. Two-way repeated measures ANOVA followed by Bonferroni test. c Representative fluorescence photomicrograph shows GFP expression in the TG 7 days after intra-TG infusion of lentivirus vector. d Real-time PCR assay of Cxcl13 shows that pretreatment with LV-Cxcl13 shRNA inhibited pIONL-induced Cxcl13 upregulation. **P < 0.01, ***P < 0.001. Student’s t test
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Related In: Results  -  Collection

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Fig3: Inhibition of CXCL13 by shRNA lentivirus attenuated pIONL-induced mechanical allodynia. a Pretreatment with LV-Cxcl13 shRNA increased the contact time between 3 and 21 days after the operation, compared to LV-NC-injected mice. Intra-TG injection was performed 7 days before pIONL (arrow). *P < 0.05, **P < 0.01, ***P < 0.001. Two-way repeated measures ANOVA followed by Bonferroni test. b Posttreatment with LV-Cxcl13 shRNA increased the contact time between 7 and 14 days after the operation, compared to LV-NC treatment. Intra-TG injection was performed 3 days after pIONL (arrow). *P < 0.05, **P < 0.01. Two-way repeated measures ANOVA followed by Bonferroni test. c Representative fluorescence photomicrograph shows GFP expression in the TG 7 days after intra-TG infusion of lentivirus vector. d Real-time PCR assay of Cxcl13 shows that pretreatment with LV-Cxcl13 shRNA inhibited pIONL-induced Cxcl13 upregulation. **P < 0.01, ***P < 0.001. Student’s t test
Mentions: To examine the role of CXCL13 in the development and maintenance of trigeminal neuropathic pain, we injected Cxcl13 shRNA lentivirus vectors (LV-Cxcl13 shRNA) into the TG before or after pIONL. Our previous study has shown that LV-Cxcl13 shRNA effectively decreased CXCL13 expression both in vitro and in vivo [11]. As shown in Fig. 3a, compared to control lentivirus injection (LV-NC), pretreatment with LV-Cxcl13 shRNA (7 days before pIONL) inhibited pIONL-induced mechanical allodynia for more than 21 days (P < 0.001, two-way repeated measures ANOVA). In addition, posttreatment with LV-Cxcl13 shRNA (3 days after pIONL) also attenuated mechanical allodynia from day 7 to day 14 (P < 0.001, two-way repeated measures ANOVA, Fig. 3b). These data suggest that CXCL13 plays an important role in both development and maintenance of trigeminal neuropathic pain.Fig. 3

Bottom Line: The effect of shRNA targeting on CXCL13 or CXCR5 on pain hypersensitivity was checked by behavioral testing. pIONL induced persistent mechanical allodynia and increased the expression of ATF3, CXCL13, and CXCR5 in the TG.Furthermore, MEK inhibitor (PD98059) attenuated mechanical allodynia and reduced TNF-α and IL-1β upregulation induced by pIONL.Pretreatment with PD98059, Etanercept, or Diacerein partially blocked CXCL13-induced mechanical allodynia, and PD98059 also reduced CXCL13-induced TNF-α and IL-1β upregulation.

View Article: PubMed Central - PubMed

Affiliation: Pain Research Laboratory, Institute of Nautical Medicine, Jiangsu Key Laboratory of Inflammation and Molecular Drug Target, Nantong University, Seyuan Road, Nantong, Jiangsu, 226019, China.

ABSTRACT

Background: Trigeminal nerve damage-induced neuropathic pain is a severely debilitating chronic orofacial pain syndrome. Spinal chemokine CXCL13 and its receptor CXCR5 were recently demonstrated to play a pivotal role in the pathogenesis of spinal nerve ligation-induced neuropathic pain. Whether and how CXCL13/CXCR5 in the trigeminal ganglion (TG) mediates orofacial pain are unknown.

Methods: The partial infraorbital nerve ligation (pIONL) was used to induce trigeminal neuropathic pain in mice. The expression of ATF3, CXCL13, CXCR5, and phosphorylated extracellular signal-regulated kinase (pERK) in the TG was detected by immunofluorescence staining and western blot. The effect of shRNA targeting on CXCL13 or CXCR5 on pain hypersensitivity was checked by behavioral testing.

Results: pIONL induced persistent mechanical allodynia and increased the expression of ATF3, CXCL13, and CXCR5 in the TG. Inhibition of CXCL13 or CXCR5 by shRNA lentivirus attenuated pIONL-induced mechanical allodynia. Additionally, pIONL-induced neuropathic pain and the activation of ERK in the TG were reduced in Cxcr5 (-/-) mice. Furthermore, MEK inhibitor (PD98059) attenuated mechanical allodynia and reduced TNF-α and IL-1β upregulation induced by pIONL. TNF-α inhibitor (Etanercept) and IL-1β inhibitor (Diacerein) attenuated pIONL-induced orofacial pain. Finally, intra-TG injection of CXCL13 induced mechanical allodynia, increased the activation of ERK and the production of TNF-α and IL-1β in the TG of WT mice, but not in Cxcr5 (-/-) mice. Pretreatment with PD98059, Etanercept, or Diacerein partially blocked CXCL13-induced mechanical allodynia, and PD98059 also reduced CXCL13-induced TNF-α and IL-1β upregulation.

Conclusions: CXCL13 and CXCR5 contribute to orofacial pain via ERK-mediated proinflammatory cytokines production. Targeting CXCL13/CXCR5/ERK/TNF-α and IL-1β pathway in the trigeminal ganglion may offer effective treatment for orofacial neuropathic pain.

No MeSH data available.


Related in: MedlinePlus