mTOR pathway is involved in ADP-evoked astrocyte activation and ATP release in the spinal dorsal horn in a rat neuropathic pain model

Abstract

Background: ATP/ADP-evoked spinal astrocyte activation plays a vital role in the development of neuropathic pain. We aim to investigate the role of mammalian target of rapamycin (mTOR) pathway on the spinal astrocyte activation in the neuropathic pain development in rats.

Methods: Sprague Dawley (SD) rats were subjected to chronic constriction of the sciatic nerve (CCI). Rapamycin or ADP was intrathecally injected daily to explore their effects on spinal astrocyte activation and pain development. Expression of glial fibrillary acidic protein (GFAP) and mTOR in the spinal dorsal horn was assessed by immunohistochemistry. Von Frey hairs and Hargreaves paw withdrawal test were conducted to evaluate mechanical allodynia and thermal sensitivity, respectively. Firefly luciferase ATP assay was used to assess the change of ATP level in cerebrospinal fluid (CSF) and medium of cultured astrocytes.

Results: GFAP expression was enhanced in the ipsilateral spinal dorsal horn from day 3 after surgery. GFAP and mTOR expression in the rat spinal dorsal horn on post-surgical day 14 was enhanced by daily intrathecal injection of ADP, which was inhibited by rapamycin. Rapamycin decreased lower mechanical pain threshold and the thermal withdrawal latency. Intrathecal injection of ADP enhanced the ATP release, which was partially inhibited by rapamycin. Study of cultured astrocytes indicated that ATP could be released from astrocytes.

Conclusion: Our data demonstrated that ADP enhanced neuropathic pain in CCI rats, which was inhibited by rapamycin. This study indicates that targeting mTOR pathway could serve as a novel therapeutic strategy in neuropathic pain management.

Keywords: adenosine diphosphate; adenosine triphosphate; astrocyte; mammalian target of rapamycin; neuropathic pain.