破骨细胞活化过程中免疫内酯LY267108对核因子κB的抑制

背景：目前尚无理想药物可用于人工关节无菌性松动的防治。研究表明红霉素对假体周围骨溶解具有较强的抑制作用，然而其抗菌活性却限制了其在人工关节松动防治中的应用。免疫内酯 LY267108是一种新型红霉素衍生物，其消除了抗菌活性，同时保留了抗炎活性。 目的：评价LY267108在破骨细胞活化过程中对核因子kB的抑制作用。 方法：将RANKL、巨噬细胞-集落刺激因子加入小鼠RAW264.7细胞系建立破骨细胞诱导模型，同时分别加入不同浓度的阿仑膦酸钠、红霉素及LY267108共培养48 h，分别采用电泳迁移率分析法及蛋白免疫印迹法测定胞核内核因子κB活性和胞浆内κB抑制蛋白α含量。 结果与结论：LY267108对核因子κB具有较强的抑制活性，10 mg/L LY267108、25 mg/L红霉素与10 mg/L阿仑膦酸钠对核因子κB具有相似的抑制作用，且显著强于10 mg/L红霉素，而25 mg/L LY267108具有最强的抑制作用。10 mg/L LY267108、25 mg/L红霉素与10 mg/L阿仑膦酸钠组中胞浆内κB抑制蛋白α水平差异无显著性意义，但显著高于10 mg/L红霉素组，而25 mg/L LY267108组胞浆内κB抑制蛋白α水平最高。提示免疫内酯LY267108在破骨细胞活化过程中，对核因子kB具有较红霉素更强的抑制作用，且安全性高于阿仑膦酸钠。同时 LY267108无抗菌活性的特性，使其成为防治人工关节无菌性松动的潜在理想药物。而LY267108对κB抑制蛋白α降解的抑制作用，可能是其抑制核因子κB活化的机制之一。

Inhibitory effects of LY267108 on nuclear factor kappa B during osteoclast activation

BACKGROUND：No ideal drugs can be used in the prevention and treatment of aseptic loosening of artificial joints. Some researchs showed that erythromycin has strong inhibitory effects on periprosthetic osteolysis. Its antibacterial activity, however, limits its application in artificial joint loosening prevention. LY267108 is a new type of erythromycin derivatives, eliminates the antibacterial activities, and retains the anti-inflammatory activity.
OBJECTIVE：To evaluate inhibitory effect of LY267108 on nuclear factor kappa B during osteoclast activation.
METHODS：RANKL and macrophage colony-stimulating factor were added to RAW264.7 cellline of a mouse model induced by osteoclasts. Simultaneously, different concentrations of alendronate sodium, erythromycin and LY267108 were cocultured for 48 hours. The activity of nuclear factor kappa B and content of intracytoplasmic inhibitory subunit of nuclear factor kappa B alpha were measured by electrophoretic mobility shift assay and western blot assay.
RESULTS AND CONCLUSION：LY267108 has a strong inhibitory effect on nuclear factor kappa B. 10 mg/L LY267108, 25 mg/L erythromycin and 10 mg/L alendronate sodium had similar inhibitory effects on nuclear factor kappa B, which was obviously stronger than 10 mg/L erythromycin. However, 25 mg/L LY267108 had strongest inhibitory effects. No significant difference in intracytoplasmic inhibitory subunit of nuclear factor kappa B alpha levels was detected among 10 mg/L LY267108, 25 mg/L erythromycin and 10 mg/L alendronate sodium groups, but was stil apparently higher than 10 mg/L erythromycin group. Levels of intracytoplasmic inhibitory subunit of nuclear factor kappa B alpha were highest in the 25 mg/L LY267108 group. Results indicated that LY267108 in the process of osteoclast activation had stronger inhibitory effects on nuclear factor kappa B compared with erythromycin, and its safety was higher than alendronate sodium. Simultaneously, LY267108 did not have antimicrobial activity, and became a potential ideal drug for prevention and treatment of aseptic loosening of artificial joints. However, the inhibitory effects of LY267108 on the degradation of inhibitory subunit of nuclear factor kappa B alpha would be a mechanism of inhibiting the activation of nuclear factor kappa B.