- Authors: Jin, X. H., Wang, L. N., Zuo, J. L., Yang, J. P., Liu, S. L.
- Year: 2014
- Journal: J Neurosci Res 92 1690-702
- Applications: in vivo / siRNA / in vivo-jetPEI
100 nM of siRNA with a stabilizing 2'-FU substitution at the 3'-end were complexed with in vivo-jetPEI in a total volume of 10 µl and injected intrathecally in the lumbare region of rats. Three injections were performed at 3 consecutive days.
Previous studies have suggested that the microglial P2X7 purinoceptor is involved in the release of tumor necrosis factor-alpha (TNFalpha) following activation of toll-like receptor-4 (TLR4), which is associated with nociceptive behavior. In addition, this progress is evoked by the activation of the P2X4 purinoceptor (P2X4R). Although P2X4R is also localized within spinal microglia in the dorsal horn, little is known about its role in cancer-induced bone pain (CIBP), which is in some ways unique. With the present rat model of CIBP, we demonstrate a critical role of the microglial P2X4R in the enhanced nociceptive transmission, which is associated with TLR4 activation and secretion of brain-derived neurotrophic factor (BDNF) and TNFalpha in the dorsal horn. We assessed mechanical threshold and spontaneous pain of CIBP rats. Moreover, P2X4R small interfering RNA (siRNA) was administered intrathecally, and real-time PCR, Western blots, immunofluorescence histochemistry, and ELISA were used to detect the expression of P2X4R, TLR4, OX-42, phosphorylated-p38 MAPK (p-p38), BDNF, and TNFalpha. Compared with controls, intrathecal injection of P2X4R siRNA could prevent nociceptive behavior induced by ATP plus lipopolysaccharide and CIBP and reduce the expression of P2X4R, TLR4, p-p38, BDNF, and TNFalpha. In addition, the increase of BDNF protein in rat microglial cells depended on P2X4 receptor signaling, which is partially associated with TLR4 activation. The ability of microglial P2X4R to activate TLR4 in spinal cord leading to behavioral hypersensitivity and oversecretion of BDNF could provide an opportunity for the prevention and treatment of CIBP.