OBJECTIVE The suppressors of cytokine signaling (SOCS) proteins are physiological suppressors of cytokine signaling which have been identified as a negative feedback loop to weaken cytokine signaling. However, the underlying molecular mechanisms is unknown. This study was to investigate the role of SOCS1 in the oxygen-glucose deprivation and reoxygenation (OGDR) or LPS-induced inflammation in microglia cell line BV-2 cells. MATERIALS AND METHODS BV-2 microglial cells were used to construct inflammation model. A SOCS1 over-expression plasmid was constructed, and the SOCS1-deficient cells were generated by utilizing the CRISPR/CAS9 system. BV-2 microglial cells were pretreated with over-expression plasmid or SOCS1 CRISPR plasmid before OGDR and LPS stimulation. The effect of SOCS1 on proinflammatory cytokines, toll-like receptor 4 (TLR4), and reactive oxygen species (ROS) were evaluated. RESULTS We found that SOCS1 increased in OGDR or LPS-treated BV-2 microglial cells in vitro. SOCS1 over-expression significantly reduced the production of proinflammatory cytokines including tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6, and CRISPR/CAS9-mediated SOCS1 knockout reversed this effect. Also we determined that SOCS1 over-expression reduced the level of reactive oxygen species (ROS) while the absence of SOCS1 increased the production of ROS after OGDR or LPS-stimulated inflammation. Furthermore, we found that OGDR and LPS induced the expression of toll-like receptor 4 (TLR4) in BV2 cells. Nevertheless, SOCS1 over-expression attenuated the expression of TLR4, while knockdown of SOCS1 upregulated TLR4. CONCLUSIONS Our study indicated that SOCS1 played a protective role under inflammatory conditions in OGDR or LPS treated BV-2 cells through regulating ROS and TLR4. These data demonstrated that SOCS1 served as a potential therapeutic target to alleviate inflammation after ischemic stroke.

中文翻译：

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细胞因子信号转导抑制剂 (SOCS)-1 通过调节 BV2 细胞中的 ROS 和 TLR4 来抑制神经炎症。

目的 细胞因子信号转导抑制蛋白 (SOCS) 蛋白是细胞因子信号转导的生理抑制因子，已被确定为削弱细胞因子信号转导的负反馈回路。然而，潜在的分子机制是未知的。本研究旨在研究 SOCS1 在小胶质细胞系 BV-2 细胞氧-葡萄糖剥夺和复氧 (OGDR) 或 LPS 诱导的炎症中的作用。材料与方法 BV-2小胶质细胞用于构建炎症模型。构建了 SOCS1 过表达质粒，利用 CRISPR/CAS9 系统生成了 SOCS1 缺陷细胞。在 OGDR 和 LPS 刺激之前，用过表达质粒或 SOCS1 CRISPR 质粒预处理 BV-2 小胶质细胞。SOCS1 对促炎细胞因子、Toll 样受体 4 (TLR4) 的影响，和活性氧 (ROS) 进行了评估。结果 我们发现 SOCS1 在体外 OGDR 或 LPS 处理的 BV-2 小胶质细胞中增加。SOCS1 过表达显着减少了促炎细胞因子的产生，包括肿瘤坏死因子 α (TNF-α)、白细胞介素 1β (IL-1β) 和 IL-6，而 CRISPR/CAS9 介导的 SOCS1 敲除逆转了这种作用。我们还确定 SOCS1 过度表达降低了活性氧 (ROS) 的水平，而 SOCS1 的缺失增加了 OGDR 或 LPS 刺激的炎症后 ROS 的产生。此外，我们发现 OGDR 和 LPS 诱导 BV2 细胞中 toll 样受体 4 (TLR4) 的表达。然而，SOCS1 过表达减弱了 TLR4 的表达，而 SOCS1 的敲低上调了 TLR4。结论 我们的研究表明，SOCS1 通过调节 ROS 和 TLR4，在 OGDR 或 LPS 处理的 BV-2 细胞中在炎症条件下发挥保护作用。这些数据表明 SOCS1 可作为减轻缺血性中风后炎症的潜在治疗靶点。