Liposome-encapsulated clodronate specifically depletes spinal microglia and reduces initial neuropathic pain

Liposome-encapsulated clodronate (LEC) is a specific depletor of macrophages. Our study characterized the LEC depletory effects, given intrathecally, on spinal microglia and assessed its effects on initiation and maintenance of neuropathic pain. Measured by using the MTT assay, LEC treatment specifically inhibited cell viability of cultured primary microglia, but not astrocytes or neurons, from neonatal rats, with an IC₅₀ of 43 μg/mL. In spinal nerve ligation-induced neuropathic rats, pretreatment (1 day but not 5 days earlier) with intrathecal LEC specifically depleted microglia (but not astrocytes or neurons) in both contralateral and ipsilateral dorsal horns by the same degree (63% vs. 71%). Intrathecal injection of LEC reversibly blocked the antinociceptive effects of the GLP-1 receptor agonist exenatide and dynorphin A stimulator bulleyaconitine, which have been claimed to be mediated by spinal microglia, whereas it failed to alter morphine- or the glycine receptor agonist gelsemine-induced mechanical antiallodynia which was mediated via the neuronal mechanisms. Furthermore, intrathecal LEC injection significantly attenuated initial (one day after nerve injury) but not existing (2 weeks after nerve injury) mechanical allodynia. Our study demonstrated that LEC, given intrathecally, is a specific spinal microglial inhibitor and significantly reduces initiation but not maintenance of neuropathic pain, highlighting an opposite role of spinal microglia in different stages of neuropathic pain.