Section of Rheumatology, Department of Biochemistry, Rush University Medical Center, Chicago, Illinois 60612, USA.
J Cell Physiol. 2013 Sep;228(9):1884-96. doi: 10.1002/jcp.24350.
The catabolic cytokine interleukin-1 (IL-1) and endotoxin lipopolysaccharide (LPS) are well-known inflammatory mediators involved in degenerative disc disease, and inhibitors of IL-1 and LPS may potentially be used to slow or prevent disc degeneration in vivo. Here, we elucidate the striking anti-catabolic and anti-inflammatory effects of bovine lactoferricin (LfcinB) in the intervertebral disc (IVD) via antagonism of both IL-1 and LPS-mediated catabolic activity using in vitro and ex vivo analyses. Specifically, we demonstrate the biological counteraction of LfcinB against IL-1 and LPS-mediated proteoglycan (PG) depletion, matrix-degrading enzyme production, and enzyme activity in long-term (alginate beads) and short-term (monolayer) culture models using bovine and human nucleus pulposus (NP) cells. LfcinB significantly attenuates the IL-1 and LPS-mediated suppression of PG production and synthesis, and thus restores PG accumulation and pericellular matrix formation. Simultaneously, LfcinB antagonizes catabolic factor mediated induction of multiple cartilage-degrading enzymes, including MMP-1, MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5, in bovine NP cells at both mRNA and protein levels. LfcinB also suppresses the catabolic factor-induced stimulation of oxidative and inflammatory factors such as iNOS, IL-6, and toll-like receptor-2 (TLR-2) and TLR-4. Finally, the ability of LfcinB to antagonize IL-1 and LPS-mediated suppression of PG is upheld in an en bloc intradiscal microinjection model followed by ex vivo organ culture using both mouse and rabbit IVD tissue, suggesting a potential therapeutic benefit of LfcinB on degenerative disc disease in the future.
分解代谢细胞因子白细胞介素-1(IL-1)和内毒素脂多糖(LPS)是已知的参与退行性椎间盘疾病的炎症介质,IL-1 和 LPS 的抑制剂可能潜在地用于减缓或预防体内椎间盘的退化。在这里,我们通过体外和离体分析阐明了牛乳铁蛋白(LfcinB)通过拮抗 IL-1 和 LPS 介导的分解代谢活性,对椎间盘(IVD)具有惊人的抗分解代谢和抗炎作用。具体而言,我们证明了 LfcinB 对 IL-1 和 LPS 介导的蛋白聚糖(PG)耗竭、基质降解酶产生和牛和人椎间盘核(NP)细胞的短期(单层)和长期(藻酸盐珠)培养模型中的酶活性的生物学拮抗作用。LfcinB 显著减弱了 IL-1 和 LPS 介导的 PG 产生和合成的抑制作用,从而恢复了 PG 的积累和细胞外基质的形成。同时,LfcinB 拮抗了多种软骨降解酶,包括 MMP-1、MMP-3、MMP-13、ADAMTS-4 和 ADAMTS-5,在牛 NP 细胞中的基因和蛋白水平上,均由分解代谢因子介导诱导。LfcinB 还抑制了分解代谢因子诱导的氧化和炎症因子的刺激,如 iNOS、IL-6 和 Toll 样受体-2(TLR-2)和 TLR-4。最后,LfcinB 拮抗 IL-1 和 LPS 介导的 PG 抑制的能力在使用小鼠和兔 IVD 组织的离体椎间盘内微注射模型和随后的离体器官培养中得到维持,这表明 LfcinB 在未来对退行性椎间盘疾病具有潜在的治疗益处。