Abstract：
To develop novel bovine lactoferrin (bLF) peptides targeting bLF-tumour necrosis factor (TNF) receptor-associated factor 6 (TRAF6) binding sites, we identified two peptides that could target bLF-TRAF6 binding sites using structural analysis. Moreover, another peptide that could bind to the TRAF6 dimerization area was selected from the bLF sequence. The effects of each peptide on cytokine expression in lipopolysaccharide (LPS)-stimulated osteoblasts (ST2) and on osteoclastogenesis were examined using an LPS-treated co-culture of primary bone marrow cells (BMCs) with ST2 cells and a single culture of osteoclast precursor cells (RAW-D) treated with soluble receptor activator of NF-κB ligand. Finally, the effectiveness of these peptides against LPS-induced alveolar bone destruction was assessed. Two of the three peptides significantly suppressed LPS-induced TNF-α and interleukin-1β expression in ST2 cells. Additionally, these peptides inhibited and reversed LPS-induced receptor activator of NF-κB ligand (RANKL) upregulation and osteoprotegerin (OPG) downregulation, respectively. Furthermore, both peptides significantly reduced LPS-induced osteoclastogenesis in the BMC-ST2 co-culture and RANKL-induced osteoclastogenesis in RAW-D cells. In vivo, topical application of these peptides significantly reduced the osteoclast number by downregulating RANKL and upregulating OPG in the periodontal ligament. It is indicated that the novel bLF peptides can be used to treat periodontitis-associated bone destruction.
摘要：
开发针对bLF-肿瘤坏死因子（TNF）受体相关因子6（TRAF6）结合位点的新型牛乳铁蛋白（bLF）肽，我们使用结构分析鉴定了两种可以靶向bLF-TRAF6结合位点的肽.此外，可以结合TRAF6二聚化区的另一种肽选自bLF序列。使用LPS处理的原代骨髓细胞（BMC）与ST2细胞的共培养和用NF-κB配体的可溶性受体激活剂处理的破骨细胞前体细胞（RAW-D）的单一培养物，检查了每种肽对脂多糖（LPS）刺激的成骨细胞（ST2）中细胞因子表达和破骨细胞生成的影响。最后,评估了这些肽对LPS诱导的牙槽骨破坏的有效性。三种肽中的两种显着抑制了LPS诱导的ST2细胞中TNF-α和白介素-1β的表达。此外,这些肽抑制和逆转LPS诱导的NF-κB受体激活剂配体（RANKL）上调和骨保护素（OPG）下调，分别。此外，两种肽均显着降低了BMC-ST2共培养物中LPS诱导的破骨细胞生成和RAW-D细胞中RANKL诱导的破骨细胞生成。在体内,这些肽的局部应用通过下调RANKL和上调牙周膜中的OPG而显着减少了破骨细胞的数量。表明新的bLF肽可用于治疗牙周炎相关的骨破坏。
