Electroacupuncture inhibits excessive interferon-γ evoked up-regulation of P2X4 receptor in spinal microglia in a CCI rat model for neuropathic pain

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Electroacupuncture inhibits excessive interferon-γ evoked up-regulation of P2X4 receptor in spinal microglia in a CCI rat model for neuropathic pain X.-M. Chen, J. Xu, J.-G. Song, B.-J. Zheng, X.-R. Wang British Journal of Anaesthesia Volume 114, Issue 1, Pages 150-157 (January 2015) DOI: 10.1093/bja/aeu199 Copyright © 2015 The Author(s) Terms and Conditions

Fig 1 PWTs for von Frey test in rats with chronic sciatic nerve constriction. (a) ipsilateral; (b) contralateral paw. EA significantly reduced allodynia shown by raised ipsilateral PWT. Mean (sd), n=12. British Journal of Anaesthesia 2015 114, 150-157DOI: (10.1093/bja/aeu199) Copyright © 2015 The Author(s) Terms and Conditions

Fig 2 P2X4R expression in spinal dorsal horn from rats undergoing chronic sciatic nerve constriction. (a) Representative photomicrographs of immunofluorescence of P2X4R (red) and OX42 (green) on the ipsilateral spinal dorsal horn on Day 14 post-nerve injury. (b) Real-time PCR showing relative mRNA level of P2X4R in spinal dorsal horn ipsilateral to injured nerve post-surgery (individual data points shown, n=4 for each group). (c) Representative western blot of P2X4R protein detected by anti-P2X4R antibody and relative level in the spinal cord ipsilateral to the nerve injury post surgery (individual data points shown, n=4 for each group). British Journal of Anaesthesia 2015 114, 150-157DOI: (10.1093/bja/aeu199) Copyright © 2015 The Author(s) Terms and Conditions

Fig 3 IFN-γ expression in spinal dorsal horn from rats undergoing chronic sciatic nerve constriction. (a) Real-time PCR (n=4) and (b). IFN-γ measured by enzyme immunoassay (n=5) on different days post-surgery (individual data points shown). British Journal of Anaesthesia 2015 114, 150-157DOI: (10.1093/bja/aeu199) Copyright © 2015 The Author(s) Terms and Conditions

Fig 4 P2X4R expression in cultured microglia. (a) Representative photomicrographs of immunofluorescence in microglia (P2X4R, red; OX42, green) with or without IFN-γ treatment for 24 h. Percentage of P2X4R+ cells are shown on the right. (b) Representative western blot (right) and relative expression (left) of P2X4R in cultured cells with or without IFN-γ treatment. Individual data points shown, n=3 for each group. **P<0.01. British Journal of Anaesthesia 2015 114, 150-157DOI: (10.1093/bja/aeu199) Copyright © 2015 The Author(s) Terms and Conditions

Fig 5 Effect of EA on rats after spinal administration of IFN-γ. (a) von Frey PWT. Mean (sd), n=10 for each group. (b) Representative photomicrographs of immunofluorescence in spinal microglia in rats on Day 7 after intrathecal injection of IFN-γ. P2X4R (red) and OX42 (green); scale bar, 100 μm. (c) Representative western blot (top) and relative P2X4R expression (bottom, n=4) in spinal dorsal horn on different days after IFN-γ administration. Individual data points shown. British Journal of Anaesthesia 2015 114, 150-157DOI: (10.1093/bja/aeu199) Copyright © 2015 The Author(s) Terms and Conditions