The present study aimed to investigate the effects of high‐mobility group box‐1 (HMGB‐1) on mechanical pain hypersensitivity and the underlying mechanism. Mouse primary astrocytes were isolated and treated as specified. A CCK‐8 assay was used to determine cytotoxicity and a gap junctional communication assay was performed. Ethidium bromide (EtBr) uptake was used to evaluate the hemichannel activity of primary astrocytes. A mouse model of neuropathic pain was developed and paw withdrawal threshold was used to evaluate hind paw sensitivity. RT‐qPCR and Western blot were used to determine mRNA and protein expression of genes, respectively. ELISA was used to measure the release of inflammatory cytokines. Treatment with HMGB‐1 increased the expression of both toll‐like receptor‐4 (TLR‐4) and connexin 43 (Cx43) in mouse primary astrocytes. HMGB‐1 also promoted gap junctional intercellular communication and hemichannel function. Our results also demonstrated that HMGB‐1‐regulated Cx43 through the JNK signaling pathway, and Cx43 was involved in HMGB‐1‐mediated inflammation in astrocytes. In vivo analysis supported the idea that HMGB‐1‐induced mechanical hypersensitivity was associated with Cx43. We therefore conclude that HMGB‐1‐induced mechanical pain hypersensitivity occurs through modulating astrocytic Cx43 via the TLR‐4/JNK signaling pathway.

中文翻译：

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High-mobility group box-1 通过 toll 样受体-4/JNK 信号通路通过星形细胞连接蛋白 43 诱导机械疼痛超敏反应。

本研究旨在研究高迁移率族框 1 (HMGB-1) 对机械性疼痛超敏反应的影响及其潜在机制。按规定分离和处理小鼠原代星形胶质细胞。使用 CCK-8 测定来确定细胞毒性，并进行间隙连接通讯测定。溴化乙锭 (EtBr) 摄取用于评估原代星形胶质细胞的半通道活性。开发了神经性疼痛的小鼠模型，并使用缩爪阈值来评估后爪敏感性。RT-qPCR 和蛋白质印迹分别用于确定基因的 mRNA 和蛋白质表达。ELISA用于测量炎性细胞因子的释放。HMGB-1 治疗增加了小鼠原代星形胶质细胞中 toll 样受体 4 (TLR-4) 和连接蛋白 43 (Cx43) 的表达。HMGB-1 还促进间隙连接细胞间通讯和半通道功能。我们的结果还表明 HMGB-1 通过 JNK 信号通路调节 Cx43，而 Cx43 参与 HMGB-1 介导的星形胶质细胞炎症。体内分析支持 HMGB-1 诱导的机械超敏反应与 Cx43 相关的观点。因此，我们得出结论，HMGB-1 诱导的机械疼痛超敏反应是通过 TLR-4/JNK 信号通路调节星形胶质细胞 Cx43 发生的。体内分析支持 HMGB-1 诱导的机械超敏反应与 Cx43 相关的观点。因此，我们得出结论，HMGB-1 诱导的机械疼痛超敏反应是通过 TLR-4/JNK 信号通路调节星形胶质细胞 Cx43 发生的。体内分析支持 HMGB-1 诱导的机械超敏反应与 Cx43 相关的观点。因此，我们得出结论，HMGB-1 诱导的机械疼痛超敏反应是通过 TLR-4/JNK 信号通路调节星形胶质细胞 Cx43 发生的。