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目的在前期研究发现免疫调控蛋白Tim-3能负性调控炎症反应的基础上,探讨该分子能否通过调控免疫及炎症细胞稳态抑制百草枯(paraquat,PQ)中毒导致的急性炎症性损伤,以期为该类中毒性疾病的防治提供新的理论依据。方法制备PQ中毒小鼠模型,首先利用Tim-3转基因小鼠观察Tim-3高表达对PQ中毒后组织损伤及炎症介质、PQ血清浓度变化的影响;其次,制备Tim-3融合蛋白(s Tim-3),观察用该融合蛋白阻断Tim-3通路对PQ中毒后组织损伤及炎症介质变化的影响并探讨其机制。结果 PQ中毒后Tim3-转基因小鼠较野生型小鼠表现出肺及肾损伤减轻,同时Tim3-转基因小鼠肺内炎症介质TNF-α、IL-1β、IL-6的水平较野生型低。监测血清中的PQ浓度发现,Tim-3转基因小鼠血清中的PQ浓度显著低于对照组。而用Tim-3-trx融合蛋白阻断Tim-3信号后,PQ中毒小鼠的免疫反应及炎症损伤进一步加重。体外机制研究发现,Tim-3信号在减轻巨噬细胞炎症介质表达、维持炎症细胞稳态中发挥关键作用。结论在PQ中毒中,Tim-3对维持炎症反应稳态发挥关键作用,深入认识该信号通路的调控机制可望为减轻中毒后急性免疫组织损伤、降低患者死亡率提供新的策略。
OBJECTIVE: In the previous study, we found that Tim-3 could negatively regulate the inflammatory response and explore whether this molecule can inhibit the acute inflammatory injury caused by paraquat (PQ) poisoning through the regulation of immunity and inflammatory cell homeostasis, With a view to provide a new theoretical basis for the prevention and treatment of such toxic diseases. Methods The model of PQ poisoning mice was established. Tim-3 transgenic mice were used to observe the effects of Tim-3 overexpression on tissue injury, inflammatory mediators and serum PQ concentration after PQ poisoning. Secondly, Tim-3 fusion protein -3). The effect of blocking Tim-3 pathway on tissue damage and inflammatory mediators after PQ poisoning was observed by using the fusion protein and its mechanism. Results The expression of TNF-α, IL-1β and IL-6 in the lung tissue of Tim3-transgenic mice was lower than that of the wild-type mice after PQ poisoning. Monitoring PQ concentrations in serum revealed that the PQ concentration in serum of Tim-3 transgenic mice was significantly lower than that of the control group. However, Tim-3-trx fusion protein blocking Tim-3 signal, PQ poisoning mice immune response and inflammatory injury further aggravated. In vitro mechanism study found that Tim-3 signal plays a key role in alleviating the expression of inflammatory mediators in macrophages and maintaining the homeostasis of inflammatory cells. Conclusion In PQ poisoning, Tim-3 plays a key role in maintaining homeostasis of inflammatory response. To understand the regulatory mechanism of this signaling pathway is expected to provide a new strategy for alleviating the acute immune tissue damage after poisoning and reducing the mortality of patients.