Ulinastatin (UTI) is a broad-spectrum serine protease inhibitor isolated and purified from human urine with strong anti-inflammatory and cytoprotective actions, which is widely used for the treatment of various diseases, such as pancreatitis and sepsis. Although the therapeutic effects of UTI are reported to be associated with a variety of mechanisms, the signaling pathways mediating the anti-inflammatory action of UTI remain to be elucidated. In the present study we carried out a systematic study on the anti-inflammatory and anti-oxidative mechanisms of UTI and their relationships in LPS-treated RAW264.7 cells. Pretreatment with UTI (1000 and 5000 U/mL) dose-dependently decreased the mRNA levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, iNOS) and upregulated anti-inflammatory cytokines (IL-10 and TGF-β1) in LPS-treated RAW264.7 cells. UTI pretreatment significantly inhibited the nuclear translocation of NF-κB by preventing the degradation of IκB-α. UTI pretreatment only markedly inhibited the phosphorylation of JNK at Thr183, but it did not affect the phosphorylation of JNK at Tyr185, ERK-1/2 and p38 MAPK; JNK was found to function upstream of the IκB-α/NF-κB signaling pathway. Furthermore, UTI pretreatment significantly suppressed LPS-induced ROS production by activating PI3K/Akt pathways and the nuclear translocation of Nrf2 via promotion of p62-associated Keap1 degradation. However, JNK was not involved in mediating the anti-oxidative stress effects of UTI. In summary, this study shows that UTI exerts both anti-inflammatory and anti-oxidative effects by targeting the JNK/NF-κB and PI3K/Akt/Nrf2 pathways.

中文翻译：

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乌司他丁通过抑制JNK /NF-κB信号通路和激活PI3K / Akt / Nrf2通路来减轻LPS诱导的小鼠巨噬细胞RAW264.7细胞的炎症。

乌司他丁（UTI）是一种从人尿液中分离纯化的广谱丝氨酸蛋白酶抑制剂，具有很强的抗炎和细胞保护作用，广泛用于治疗各种疾病，例如胰腺炎和败血症。尽管据报道UTI的治疗作用与多种机制有关，但仍需要阐明介导UTI抗炎作用的信号传导途径。在本研究中，我们对UTI的抗炎和抗氧化机制及其在LPS处理的RAW264.7细胞中的关系进行了系统的研究。用UTI（1000和5000 U / mL）进行预处理可剂量依赖性地降低促炎细胞因子（IL-1β，IL-6，TNF-α，iNOS）和抗炎细胞因子（IL-10和TGF）的mRNA水平-β1）在LPS处理的RAW264.7细胞中。UTI预处理可通过阻止IκB-α的降解来显着抑制NF-κB的核易位。UTI预处理仅显着抑制Thr183处JNK的磷酸化，但不影响Tyr185，ERK-1 / 2和p38 MAPK处JNK的磷酸化。发现JNK在IκB-α/NF-κB信号传导途径的上游起作用。此外，UTI预处理通过激活PI3K / Akt途径以及通过促进p62相关的Keap1降解促进Nrf2的核转运，从而显着抑制LPS诱导的ROS产生。但是，JNK不参与介导UTI的抗氧化应激作用。总之，这项研究表明，UTI通过靶向JNK /NF-κB和PI3K / Akt / Nrf2途径发挥抗炎和抗氧化作用。