Intrathecal Injection of JWH-015 Attenuates Bone Cancer Pain Via Time-Dependent Modification of Pro-inflammatory Cytokines Expression and Astrocytes Activity in Spinal Cord

Abstract

Cannabinoid receptor type 2 (CB2) agonists display potential analgesic effects in acute and neuropathic pain. However, its complex cellular and molecular mechanisms in bone cancer pain remain unclear. And less relevant reports concerned its time-dependent effects on the long-lasting modifications of behavior, spinal inflammatory cytokines levels, astrocytes activity induced by bone cancer pain. A rat model of bone cancer pain induced by intra-tibia inoculation of Walker 256 mammary gland carcinoma cells was utilized. Pain behaviors at different time points were assessed by ambulatory pain scores and paw withdrawal mechanical threshold (PWMT). Pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, IL-18, and tumor necrosis factor alpha (TNF-α), were quantitated by Western blots. Glial activity was assessed by immunohistochemistry. Intra-tibia inoculation of Walker 256 mammary gland carcinoma cells induced progressive bone cancer pain; a long-term up-regulation of IL-1β, IL-6, IL-18, and TNF-α; and the activation of glia in spinal cord. Activation of microglia was first evident on day 4 after surgery and reached to a peak on day 7 while activation of astrocytes was on day 10. A single intrathecal injection of JWH-015 attenuated bone cancer induced spontaneous pain and mechanical allodynia, reduced the expression of pro-inflammatory cytokines, and inhibited the activity of astrocytes. All the modifications were transient and peaked at 24 h after JWH-015 administration. Furthermore, the protective effects of JWH-015 were reversed in the presence of CB2-selective antagonist AM630. Overall, our results provided evidences for the persistent participation of inflammation reaction in the progression of bone cancer pain, and demonstrated that JWH-015 reduced the expression of IL-1β, IL-6, IL-18, and TNF-α and inhibited astrocytes activation in a time-dependent manner, thereby displaying an analgesic effect.

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Abbreviations

CB1 :

Cannabinoid receptor type 1

CB2 :

Cannabinoid receptor type 2

IL:

Interleukin

TNF-α:

Tumor necrosis factor alpha

PWMT:

Paw withdrawal mechanical threshold

CNS:

Central nervous system

DMSO:

Dimethyl sulfoxide

PBS:

Phosphate buffered saline

GFAP:

Glial fibrillary acidic protein

SD:

Standard deviation

CCR:

Chemokine receptor

COX:

Cyclo-oxygen-ase

NMDA:

N-methyl-d-aspartic acid receptor

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Acknowledgments

This research was supported by the National Natural Science Foundation of China (81070892, 81171047, 81171048, and 81371207) and a grant from the Department of Health of Jiangsu Province of China (XK201140, RC2011006).

Conflict of Interest

The authors declare that they have no conflicts of interest.

Authors’ Contributions

All of the authors read and approved the final manuscript. CEL made substantial contributions to the experiments. YL was mainly involved in the pain behavioral tests and the spinal astrocyte activity assay. BS and YZ performed the surgical procedure, administration of drugs, and Western blotting studies. YES and BLH were responsible for statistical analyses. All of these individuals participated in drafting the manuscript. XPG and ZLM conceived the idea, designed the study, and helped to revise the manuscript.

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Correspondence to Zhengliang Ma or Xiaoping Gu.

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Cui’e Lu, Yue Liu, Bei Sun and Yu’e Sun contributed equally to this work.

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Lu, C., Liu, Y., Sun, B. et al. Intrathecal Injection of JWH-015 Attenuates Bone Cancer Pain Via Time-Dependent Modification of Pro-inflammatory Cytokines Expression and Astrocytes Activity in Spinal Cord. Inflammation 38, 1880–1890 (2015). https://doi.org/10.1007/s10753-015-0168-3

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KEY WORDS

  • bone cancer pain
  • CB2
  • astrocytes
  • pro-inflammatory cytokines
  • time-dependent