FG2216 synthesis SGCs tightly ensheath the soma of DRG neuro
SGCs tightly ensheath the soma of DRG neurons (Costa and Moreira, 2015, Hanani, 2005, Takeda et al., 2009). Intimate associations between SGCs and neurons facilitate bidirectional regulation of SGC function and neuronal excitability. After nerve injury, neuronal hyperexcitability can result in SGC activation and thus induce neuropathic pain (Costa and Moreira, 2015, Hanani, 2005, Takeda et al., 2009, Ying et al., 2017). Elevation of the activation marker GFAP indicates activation of SGCs in DRG. The present study showed that GFAP FG2216 synthesis was increased in DRG SCGs after gp120 treatment, which suggests activation of SGCs in gp120 rats. P2Y12 is expressed in DRG SGCs (Katagiri et al., 2012, Kobayashi et al., 2013). Our results also indicated the co-expression values of P2Y12 and GFAP in the gp120 group were increased compared with those in the sham group. Thus, the P2Y12 receptor is involved in the transmission of signals by DRG SGCs resulting from nerve injury in gp120 rats. P2Y12 shRNA treatment in DRG SGCs decreased expression of GFAP protein and co-expression of P2Y12 and GFAP in gp120-treated rats. Simultaneously, the MWT and TWL were increased in gp120 rats after P2Y12 shRNA treatment compared with rats treated with gp120 alone. Therefore, down-regulation of the P2Y12 receptor decreased activation of DRG SGCs and relieved pain behaviors in gp120-treated rats. Activation of DRG SGCs can increase the release of proinflammatory cytokines. Enhanced levels of proinflammatory cytokines augment abnormal neuronal excitability and contribute to neuropathic pain (Kajander et al., 1992, Sachs et al., 2002, Takeda et al., 2007, Takeda et al., 2009). HIV gp120 also stimulates the nervous system to release pro-inflammatory cytokines (Hao, 2013, Yuan et al., 2014, Zheng et al., 2011). Our results indicated that up-regulation of the P2Y12 receptor in gp120-treated rats was accompanied by enhanced IL-1β protein expression. TNF-R1 is activated by TNF-α in DRG neurons, increasing neuronal excitability (Illes et al., 2012, Kajander et al., 1992). Our results also revealed that the expression levels of TNF-R1 in DRG of gp120-treated rats were enhanced compared with sham rats. Up-regulation of the IL-1β protein and TNF-R1 in DRG can lead to abnormal neuronal excitability. Proinflammatory cytokines in DRG enhanced neuronal excitability and promoted hyperalgesia in gp120-treated rats. P2Y12 shRNA treatment decreased the expression levels of the IL-1β protein and TNF-R1 in DRG and reduced hyperalgesia in gp120 rats. P2Y12 receptor activation augmented IL-1β release in neuropathic pain (Horváth et al., 2014). Thus, down-regulation of the P2Y12 receptor decreased activation of DRG SGCs, and then reduced expression of IL-1β and TNF-R1. Down-regulated expression of IL-1β and TNF-R1 lessened abnormal signal between neuron and SGC in DRG as well as relieved neuropathic pain behaviors induced by gp120 treatment. Activation of the protein kinase B/Akt signaling pathway is involved in neuropathic pain mechanisms (Guedes et al., 2008, Xu et al., 2007, Sun et al., 2006). Accordingly, inhibition of protein kinase B/Akt activation reduced pain behaviors associated with neuropathic pain (Sun et al., 2006, Xu et al., 2007). The P2Y12 receptor is involved in Akt activation (Irino et al., 2008). Our data indicated that the integrated optical density (IOD) ratio of p-Akt to Akt in gp120 rats was higher than in sham rats. It is possible that Akt phosphorylation in DRG is involved in hyperalgesia in gp120-treated rats. After P2Y12 shRNA treatment in gp120 rats, the IOD ratios of p-Akt to Akt were significantly decreased. Our studies revealed that P2Y12 shRNA treatment silenced up-regulation of the P2Y12 receptor and decreased p-Akt in DRG of gp120-treated rats. Consequently, the degree of p-Akt reduction in the gp120-treated rats was closely associated with reduction in pain behaviors after P2Y12 shRNA treatment. Therefore, we conclude that silencing the P2Y12 receptor reduced Akt activation in DRG in gp120-treated rats.