背景:炎症在慢性疾病的发展中起着关键作用。减少慢性炎症是预防和管理许多慢性疾病的重要策略。在中医中,经过加工的ButhusmartensiiKarsch(BmK)蝎子(也称为“Quanxie”)已被用于治疗慢性炎症性关节炎和脊柱炎数百年,这表明“Quanxie”可能被用作鉴定新的抗炎化合物的资源。然而,加工的BmK蝎子抗炎作用的分子基础和强调的机制仍不清楚。
目的:该研究旨在确定巨噬细胞表达的Kv1.3在加工的BmK蝎毒的抗炎作用中的潜在参与,以及鉴定源自加工过的BmK蝎子的新Kv1.3阻断剂。
方法:在本研究中,使用角叉菜胶诱导的爪水肿测定体内和体外抗炎活性,分别建立LPS诱导的脓毒症小鼠模型和LPS诱导的巨噬细胞活化模型。加工的BmK蝎水提取物的效果,在转染的HEK293细胞或小鼠BMDMs上通过全细胞电压钳记录检测不同钾通道上加工的BmK毒液和BmKK2。使用RT-PCR和竞争性酶联免疫吸附法检测细胞因子。高效液相色谱法,使用SDS-PAGE和肽质谱分析来分离和鉴定BmKK2。SiRNA,采用免疫印迹和流式细胞术分析BmKK2的抗炎机制。
结果:在这里,我们证明了BmKK2,一种靶向Kv1.3的热稳定毒素是加工过的BmK蝎中的关键抗炎成分。BmKK2通过靶向和抑制巨噬细胞Kv1.3的活性来抑制炎症,从而抑制NF-kB-NLPR3通路的激活和随后的炎症因子的释放。
结论:这些发现为“Quanxie”抗炎作用的分子基础提供了新的见解,并强调了靶向巨噬细胞上表达的Kv1.3作为抗炎方法的重要性。
BACKGROUND: Inflammation plays pivotal role in the development of chronic diseases. Reducing chronic inflammation is an important strategy for preventing and managing many chronic diseases. In traditional Chinese medicine, the processed Buthus martensii Karsch (BmK)
scorpion (also called \"Quanxie\") has been used to treat chronic inflammatory arthritis and spondylitis for hundreds of years suggests that \"Quanxie\" could potentially be utilized as a resource for identifying new anti-inflammatory compounds. However, the molecular basis and the underline mechanism for the anti-inflammatory effect of processed BmK
scorpion are still unclear.
OBJECTIVE: The study aims to determine the potential involvement of macrophage-expressed Kv1.3 in the anti-inflammatory effect of processed BmK
scorpion venom, as well as to identify new Kv1.3 blockers derived from processed BmK
scorpion.
METHODS: In this study, the in vivo and in vitro anti-inflammatory activities were determined using carrageenan-induced paw edema, LPS-induced sepsis mouse models and LPS-induced macrophage activation model respectively. The effect of processed BmK
scorpion water extract, processed BmK venom and BmKK2 on different potassium channels were detected by whole-cell voltage-clamp recordings on transfected HEK293 cells or mouse BMDMs. The cytokines were detected using Q-PCR and competitive enzyme-linked immunosorbent assay. High performance liquid chromatography, SDS-PAGE and peptide Mass Spectrometry analysis were used to isolate and identify the BmKK2. SiRNA, western blotting and flow cytometry were used to analysis the anti-inflammatory mechanism of BmKK2.
RESULTS: Here we demonstrate that BmKK2, a thermostable toxin targeting Kv1.3 is the critical anti-inflammatory component in the processed BmK scorpion. BmKK2 inhibits inflammation by targeting and inhibiting the activity of macrophage Kv1.3, thereby inhibiting the activation of NF-κB-NLRP3 pathway and the subsequent release of inflammatory factors.
CONCLUSIONS: These findings provide new insights into the molecular basis of the anti-inflammatory effects of \"Quanxie\" and highlight the importance of targeting Kv1.3 expressed on macrophages as an anti-inflammatory approach.