Background

Adenosine deaminase acting on RNA 3 (ADAR3) was known as a prognosis factor in gliomas, while its function on neuropathic pain (NP) is barely investigated. Therefore, our present study concentrated on the potential role of ADAR3 in NP.

Methods

The chronic constriction injury (CCI) mouse model was established to induce NP in vivo. Behavioral experiments were carried out to analyze mechanical allodynia and thermal hyperalgesia. RT-qPCR and western blotting assays were used to detect the mRNA and protein expressions. The ADAR3-overexpressed adenovirus was injected into the CCI mice through an intrathecal catheter. ELISA was used to detect the contents of IL (interleukin)-6, IL-10, TNF (tumor necrosis factor)-α, IL-1β and IL-18. NLR Family Pyrin Domain Containing 3 (NLRP3) was predicted to be the target gene of ADAR3 using Starbase. The interaction between ADAR3 and NLRP3 was verified via RNA pull-down, RNA immunoprecipitation and Pearson's correlation coefficient assays. Immunohistochemical staining assay visualized the expressions of NLRP3 and caspase1.

Results

Allodynia and hyperalgesia were exacerbated in the CCI mice, which implied a successful establishment of the NP model, while ADAR3 expression level was suppressed. After injecting ADAR3-overexpressed adenovirus into the CCI mice, allodynia, hyperalgesia and inflammation were all restrained. Moreover, NLRP3 was verified to negatively correlated with ADAR3. Additionally, the pyroptosis-related protein NLRP3, ASC, caspase1, IL-1β, IL-18 and GSDMD expressions were all decreased by ADAR3.

Conclusion

In conclusion, ADAR3 alleviated inflammation and pyroptosis of NP through targeting NLRP3, which suggested a therapeutical target for NP.

中文翻译：

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ADAR3通过靶向NLRP3减轻慢性收缩损伤小鼠神经性疼痛的炎症和细胞焦亡

背景

作用于 RNA 3 (ADAR3) 的腺苷脱氨酶被认为是神经胶质瘤的预后因素，而其对神经性疼痛 (NP) 的功能几乎没有研究。因此，我们目前的研究集中在 ADAR3 在 NP 中的潜在作用。

方法

建立慢性缩窄性损伤（CCI）小鼠模型以在体内诱导NP. 进行行为实验以分析机械异常性疼痛和热痛觉过敏。RT-qPCR 和蛋白质印迹分析用于检测 mRNA 和蛋白质表达。通过鞘内导管将 ADAR3 过表达的腺病毒注射到 CCI 小鼠中。采用ELISA法检测IL（白细胞介素）-6、IL-10、TNF（肿瘤坏死因子）-α、IL-1β和IL-18的含量。使用 Starbase 预测 NLR Family Pyrin Domain Containing 3 (NLRP3) 是 ADAR3 的靶基因。ADAR3 和 NLRP3 之间的相互作用通过 RNA pull-down、RNA 免疫沉淀和 Pearson 相关系数测定进行验证。免疫组织化学染色分析显示 NLRP3 和 caspase1 的表达。

结果

CCI 小鼠的异常性疼痛和痛觉过敏加剧，这意味着 NP 模型的成功建立，而 ADAR3 的表达水平受到抑制。将 ADAR3 过表达的腺病毒注射到 CCI 小鼠后，异常性疼痛、痛觉过敏和炎症均得到抑制。此外，NLRP3 被证实与 ADAR3 呈负相关。此外，焦亡相关蛋白 NLRP3、ASC、caspase1、IL-1β、IL-18 和 GSDMD 的表达均被 ADAR3 降低。

结论

总之，ADAR3 通过靶向 NLRP3 减轻了 NP 的炎症和细胞焦亡，这表明了 NP 的治疗靶点。