Journal: The journal of pain
Article Title: Microglial P2Y12 Signaling Contributes to Cisplatin-induced Pain Hypersensitivity via IL-18-mediated Central Sensitization in the Spinal Cord.
doi: 10.1016/j.jpain.2023.01.005
Figure Lengend Snippet: Figure 5. Spinal blockade of the P2Y12/Src family kinase (SFK)/p38 pathway suppresses the cisplatin-induced IL-18 production. (A) IL-18 immunoreactivity was colocalized with IBA-1, (B) but not with NeuN or GFAP in the spinal dorsal horn. (C) IL-18 immunoreac- tivity was colocalized with P2Y12, p-SFK, and p-p38 in the spinal dorsal horn. Tissues were harvested on day 15 post administration. Scale bars: 50 mm and 20 mm (zoom). (D−F) Real-time qPCR analyses showed that MRS2395 (D), PP2 (E) or SB239063 (F) significantly attenuated the cisplatin-induced upregulation of IL-18 mRNA expression in the spinal dorsal horn. (G−I) Western blotting analyses showed that MRS2395 (G), PP2 (H) or SB239063 (I) significantly attenuated the cisplatin-induced upregulation of IL-18 protein expression in the spinal dorsal horn. MRS2395 (2 mg, intrathecally), PP2 (10 mg, intrathecally), PP3 (10 mg, intrathecally) or SB239063 (10 mg, intrathecally) was injected daily once, on days 12, 13, 14, and 15 after the first cisplatin injection. Tissues were harvested 4 h following the last injection on day 15. Data are shown as the mean § SEM. **P < .01, in comparison with the saline + DMSO (or PP3) group; ##P < .01, compared with the cisplatin + DMSO (or PP3) group, n = 4 per group, one-way ANOVA with the post hoc Dunnett test. Abbreviations: ANOVA, analysis of variance; DMSO, dimethyl sulfoxide; GFAP, glial fibrillary acidic protein; IBA-1, ionized cal- cium-binding adapter molecule 1; IL-18, interleukin-18; NeuN, neuronal nuclei; qPCR, quantitative polymerase chain reaction; SEM, standard error of the mean; SFK, Src family kinase.TagedEnd
Article Snippet: Src family kinase inhibitor PP2 (S7008) and P38 inhibitor SB239063 (S7741) were obtained from Selleck Chemicals (China).
Techniques: Expressing, Western Blot, Injection, Comparison, Saline, Binding Assay, Real-time Polymerase Chain Reaction