Park Hye Ji, Lee Seong Ho, Son Dong Ju, Oh Ki Wan, Kim Ki Hyun, Song Ho Sueb, Kim Goon Joung, Oh Goo Taeg, Yoon Do Young, Hong Jin Tae
College of Pharmacy, Chungbuk National University, 48 Gaesin-dong, Heungduk-gu, Cheongju, Chungbuk 361-763, South Korea.
Arthritis Rheum. 2004 Nov;50(11):3504-15. doi: 10.1002/art.20626.
To investigate the molecular mechanisms of the antiarthritic effects of bee venom (BV) and melittin (a major component of BV) in a murine macrophage cell line (Raw 264.7) and in synoviocytes obtained from patients with rheumatoid arthritis.
We evaluated the antiarthritic effects of BV in a rat model of carrageenan-induced acute edema in the paw and in a rat model of chronic adjuvant-induced arthritis. The inhibitory effects of BV and melittin on inflammatory gene expression were measured by Western blotting, and the generation of prostaglandin E(2) (PGE(2)) and nitric oxide (NO) and the intracellular calcium level were assayed. NF-kappaB DNA binding and transcriptional activity were determined by gel mobility shift assay or by luciferase assay. Direct binding of BV and melittin to the p50 subunit of NF-kappaB was determined with a surface plasmon resonance analyzer.
BV (0.8 and 1.6 mug/kg) reduced the effects of carrageenan- and adjuvant-induced arthritis. This reducing effect was consistent with the inhibitory effects of BV (0.5, 1, and 5 mug/ml) and melittin (5 and 10 mug/ml) on lipopolysaccharide (LPS; 1 mug/ml)-induced expression of cyclooxygenase 2, cytosolic phospholipase A(2), inducible NO synthase, generation of PGE(2) and NO, and the intracellular calcium level. BV and melittin prevented LPS-induced transcriptional and DNA binding activity of NF-kappaB via the inhibition of IkappaB release and p50 translocation. BV (affinity [K(d)] = 4.6 x 10(-6)M) and melittin (K(d) = 1.2 x 10(-8)M) bound directly to p50.
Target inactivation of NF-kappaB by directly binding to the p50 subunit is an important mechanism of the antiarthritic effects of BV.
研究蜂毒（BV）及其主要成分蜂毒肽在小鼠巨噬细胞系（Raw 264.7）和类风湿关节炎患者滑膜细胞中的抗关节炎作用分子机制。
我们在角叉菜胶诱导的大鼠足急性水肿模型和慢性佐剂诱导的关节炎大鼠模型中评估了BV的抗关节炎作用。通过蛋白质印迹法检测BV和蜂毒肽对炎症基因表达的抑制作用，并检测前列腺素E2（PGE2）、一氧化氮（NO）的生成及细胞内钙水平。通过凝胶迁移率变动分析或荧光素酶测定法确定核因子κB（NF-κB）DNA结合和转录活性。用表面等离子体共振分析仪测定BV和蜂毒肽与NF-κB p50亚基的直接结合。
BV（0.8和1.6μg/kg）减轻了角叉菜胶和佐剂诱导的关节炎的影响。这种减轻作用与BV（0.5、1和5μg/ml）和蜂毒肽（5和10μg/ml）对脂多糖（LPS；1μg/ml）诱导的环氧化酶2、胞质磷脂酶A2、诱导型NO合酶表达、PGE2和NO生成以及细胞内钙水平的抑制作用一致。BV和蜂毒肽通过抑制IκB释放和p50易位，阻止了LPS诱导的NF-κB转录和DNA结合活性。BV（亲和常数[K(d)] = 4.6 x 10(-6)M）和蜂毒肽（K(d) = 1.2 x 10(-8)M）直接与p50结合。
通过直接与p50亚基结合使NF-κB靶向失活是BV抗关节炎作用的重要机制。
