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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

pIONL induces persistent CXCR5 expression in the TG. a The time course of Cxcr5 mRNA expression in the ipsilateral TG from naïve, sham, and pIONL-operated mice. pIONL increased Cxcr5 expression at 3, 10, and 21 days, compared to sham. *P < 0.05, **P < 0.01. Two-way ANOVA followed by Bonferroni test. b Western blot shows increased CXCR5 protein level 10 days after pIONL, compared to sham. *P < 0.05. One-way ANOVA followed by Bonferroni test. c Representative images of CXCR5 immunofluorescence in the TG. CXCR5-IR was low in naïve mice (c) and sham mice (d), but increased in the TG of pIONL mice (e). f–h Double staining of CXCR5 (f) and neuronal marker β-III tubulin (g) shows the neuronal expression of CXCR5
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Fig4: pIONL induces persistent CXCR5 expression in the TG. a The time course of Cxcr5 mRNA expression in the ipsilateral TG from naïve, sham, and pIONL-operated mice. pIONL increased Cxcr5 expression at 3, 10, and 21 days, compared to sham. *P < 0.05, **P < 0.01. Two-way ANOVA followed by Bonferroni test. b Western blot shows increased CXCR5 protein level 10 days after pIONL, compared to sham. *P < 0.05. One-way ANOVA followed by Bonferroni test. c Representative images of CXCR5 immunofluorescence in the TG. CXCR5-IR was low in naïve mice (c) and sham mice (d), but increased in the TG of pIONL mice (e). f–h Double staining of CXCR5 (f) and neuronal marker β-III tubulin (g) shows the neuronal expression of CXCR5

Mentions: CXCR5 was reported to be the sole receptor for CXCL13 [25]. We then checked the time course of Cxcr5 mRNA expression in the TG. pIONL induced persistent Cxcr5 mRNA upregulation, which started at day 3 and maintained at day 21 (P < 0.05 or 0.01, pIONL vs. sham, two-way ANOVA followed by Bonferroni test, Fig. 4a). Western blot showed that CXCR5 protein level was significantly increased in pIONL mice at 10 days (P < 0.05, pIONL vs. sham or naive, one-way ANOVA followed by Bonferroni test, Fig. 4b). Immunostaining further revealed that CXCR5 had low expression in the TG in naïve mice (Fig. 4c) and sham-operated mice (Fig. 4d). CXCR5-IR was markedly increased 10 days after pIONL (Fig. 4e). In addition, CXCR5 was highly colocalized with β-III tubulin (Fig. 4f–h), indicating the neuronal expression of CXCR5 in the TG.Fig. 4


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)

pIONL induces persistent CXCR5 expression in the TG. a The time course of Cxcr5 mRNA expression in the ipsilateral TG from naïve, sham, and pIONL-operated mice. pIONL increased Cxcr5 expression at 3, 10, and 21 days, compared to sham. *P < 0.05, **P < 0.01. Two-way ANOVA followed by Bonferroni test. b Western blot shows increased CXCR5 protein level 10 days after pIONL, compared to sham. *P < 0.05. One-way ANOVA followed by Bonferroni test. c Representative images of CXCR5 immunofluorescence in the TG. CXCR5-IR was low in naïve mice (c) and sham mice (d), but increased in the TG of pIONL mice (e). f–h Double staining of CXCR5 (f) and neuronal marker β-III tubulin (g) shows the neuronal expression of CXCR5
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Fig4: pIONL induces persistent CXCR5 expression in the TG. a The time course of Cxcr5 mRNA expression in the ipsilateral TG from naïve, sham, and pIONL-operated mice. pIONL increased Cxcr5 expression at 3, 10, and 21 days, compared to sham. *P < 0.05, **P < 0.01. Two-way ANOVA followed by Bonferroni test. b Western blot shows increased CXCR5 protein level 10 days after pIONL, compared to sham. *P < 0.05. One-way ANOVA followed by Bonferroni test. c Representative images of CXCR5 immunofluorescence in the TG. CXCR5-IR was low in naïve mice (c) and sham mice (d), but increased in the TG of pIONL mice (e). f–h Double staining of CXCR5 (f) and neuronal marker β-III tubulin (g) shows the neuronal expression of CXCR5
Mentions: CXCR5 was reported to be the sole receptor for CXCL13 [25]. We then checked the time course of Cxcr5 mRNA expression in the TG. pIONL induced persistent Cxcr5 mRNA upregulation, which started at day 3 and maintained at day 21 (P < 0.05 or 0.01, pIONL vs. sham, two-way ANOVA followed by Bonferroni test, Fig. 4a). Western blot showed that CXCR5 protein level was significantly increased in pIONL mice at 10 days (P < 0.05, pIONL vs. sham or naive, one-way ANOVA followed by Bonferroni test, Fig. 4b). Immunostaining further revealed that CXCR5 had low expression in the TG in naïve mice (Fig. 4c) and sham-operated mice (Fig. 4d). CXCR5-IR was markedly increased 10 days after pIONL (Fig. 4e). In addition, CXCR5 was highly colocalized with β-III tubulin (Fig. 4f–h), indicating the neuronal expression of CXCR5 in the TG.Fig. 4

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