School of Pharmacy, Jinan University, Guangzhou, 510632, Guangdong, China,
Inflammation. 2015;38(3):1374-83. doi: 10.1007/s10753-015-0109-1.
Inflammation was the important pathological process of many disease developments, but current therapeutic means for inflammatory diseases are not satisfactory. Chemokines and their receptors represent valuable targets for anti-inflammatory drug discovery. The N15P polypeptide (sequence: LGASWHRPDKCCLGY) is independently developed by our research group, it is a new CXCR4 antagonist drug derived from viral macrophage inflammatory protein-II (vMIP-II). This study aims to clarify the anti-inflammatory potency of N15P polypeptide on the lipopolysaccharide (LPS)-induced inflammation in vitro. In this study, we evaluated the anti-inflammatory effects of N15P polypeptide by the LPS-induced peripheral blood mononuclear cell (PBMC) model and measured the level of inflammatory factors (tumor necrosis factor alpha (TNF-α), IL-6, IL-8, nuclear factor kappaB (NF-κB), cyclooxygenase-2 (COX-2), Toll-like receptor 4 (TLR4), MyD88, phosphoinositide 3-kinase (PI3K), and Akt). The messenger RNA (mRNA) expressions of inflammatory factors were analyzed by real-time PCR (RT-PCR) microarray analysis, and the production of inflammatory factors was measured further by enzyme-linked immunosorbent assay (ELISA) and Western blot. The results showed that the expression of inflammatory factors (TNF-α, IL-6, IL-8, NF-κB, COX-2, TLR4, MyD88, PI3K, and Akt) was downregulated by N15P peptide, suggesting that N15P peptide has a strong inhibitory effect on the inflammatory responses induced by LPS.
炎症是许多疾病发展的重要病理过程，但目前针对炎症性疾病的治疗手段并不令人满意。趋化因子及其受体是抗炎药物发现的有价值的靶点。N15P 多肽（序列：LGASWHRPDKCCLGY）是由我们的研究团队独立开发的，它是一种源自病毒巨噬细胞炎症蛋白-II（vMIP-II）的新型 CXCR4 拮抗剂药物。本研究旨在阐明 N15P 多肽在体外脂多糖（LPS）诱导的炎症中的抗炎效力。在这项研究中，我们通过 LPS 诱导的外周血单个核细胞（PBMC）模型评估了 N15P 多肽的抗炎作用，并测量了炎症因子（肿瘤坏死因子-α（TNF-α）、IL-6、IL-8、核因子 kappaB（NF-κB）、环氧化酶-2（COX-2）、Toll 样受体 4（TLR4）、MyD88、磷酸肌醇 3-激酶（PI3K）和 Akt）的水平。通过实时 PCR（RT-PCR）微阵列分析分析了炎症因子的信使 RNA（mRNA）表达，并通过酶联免疫吸附测定（ELISA）和 Western blot 进一步测量了炎症因子的产生。结果表明，N15P 肽下调了炎症因子（TNF-α、IL-6、IL-8、NF-κB、COX-2、TLR4、MyD88、PI3K 和 Akt）的表达，表明 N15P 肽对 LPS 诱导的炎症反应具有很强的抑制作用。
