Abstract：
Due to the urgent need to create appropriate treatment techniques, which are currently unavailable, LPS-induced sepsis has become a serious concern on a global scale. The primary active component in the pathophysiology of inflammatory diseases such as sepsis is the Gram-negative bacterial lipopolysaccharide (LPS). LPS interacts with cell surface TLR4 in macrophages, causing the formation of reactive oxygen species (ROS), TNF-α, IL-1β and oxidative stress. It also significantly activates the MAPKs and NF-κB pathway. Excessive production of pro-inflammatory cytokines is one of the primary characteristic features in the onset and progression of inflammation. Cytokines mainly signal through the JAK/STAT pathway. We hypothesize that blocking of TLR4 along with TNFR1 might be beneficial in suppressing the effects of STAT1/STAT3 due to the stimulation of SOCS3 proteins. Prior to the LPS challenge, the macrophages were treated with antibodies against TLR4 and TNFR1 either individually or in combination. On analysis of the macrophage populations by flowcytometry, it was seen that receptor blockade facilitated the phenotypic shift of the M1 macrophages towards M2 resulting in lowered oxidative stress. Blocking of TLR4/TNFR1 upregulated the SOCS3 and mTOR expressions that enabled the transition of inflammatory M1 macrophages towards the anti-inflammatory M2 phenotype, which might be crucial in curbing the inflammatory responses. Also the reduction in the production of inflammatory cytokines such as IL-6, IL-1β due to the reduction in the activation of the STAT1 and STAT3 molecules was observed in our combination treatment group. All these results indicated that neutralization of both TLR4 and TNFR1 might provide new insights in establishing an alternative therapeutic strategy for LPS-sepsis.
摘要：
由于迫切需要创造适当的治疗技术，目前不可用，LPS诱导的脓毒症在全球范围内已成为严重关注的问题。炎性疾病如脓毒症的病理生理学中的主要活性成分是革兰氏阴性细菌脂多糖(LPS)。LPS与巨噬细胞中的细胞表面TLR4相互作用，导致活性氧（ROS）的形成，TNF-α,IL-1β与氧化应激。它还显著激活MAPKs和NF-κB途径。促炎细胞因子的过度产生是炎症发作和进展的主要特征之一。细胞因子主要通过JAK/STAT途径发出信号。我们假设由于SOCS3蛋白的刺激，TLR4和TNFR1的阻断可能有利于抑制STAT1/STAT3的作用。在LPS挑战之前，用针对TLR4和TNFR1的抗体单独或联合治疗巨噬细胞.通过流式细胞术分析巨噬细胞群，可以看出,受体阻断促进了M1巨噬细胞向M2的表型转变,从而降低了氧化应激。TLR4/TNFR1的阻断上调SOCS3和mTOR的表达，使得炎性M1巨噬细胞向抗炎M2表型转变，这可能对抑制炎症反应至关重要。在我们的组合治疗组中还观察到由于STAT1和STAT3分子的活化减少而导致的炎性细胞因子如IL-6、IL-1β的产生减少。所有这些结果表明，TLR4和TNFR1的中和可能为建立LPS败血症的替代治疗策略提供新的见解。
