以一氧化氮合成及第二型環氧化之誘發表現探討如意金黃膏之消炎止痛作用   全文下載

Inducible Expression of NOS and COX-2 in Evaluating the Effects of Ruyi-Jinhuang Gao on Somatic Pain – Associated Inflammation

「如意金黃散」係中國古方，屬於傷科用散劑，記載於醫宗金鑑（清朝，作者：吳謙），也是目前中醫傷科之外用散劑。本研究之「如意金黃膏」原料即「如意金黃散」。本研究對如意金黃膏進行有關抗發炎及致敏性之實驗。以西方墨點分析發現如意金黃膏可抑制脂多醣（1 μg/mL）所誘發巨噬細胞一氧化氮合成（iNOS）及第二型環氧化?（COX-2）之表現，具抗發炎作用之潛力。其植物成員則顯示不同程度抑制一氧化氮合成及第二型環氧化?之表現。其中，以甘草、薑黃、蒼朮在細胞培養15 hr之抑制效果最強。此一結果，可用於改善如意金黃散處方之參考。如意金黃膏之植物成員萃取物對於巨噬細胞在細胞培養後6及15 hr呈現不同的細胞存活率或細胞毒性；如意金黃膏本身在細胞培養後15 hr才呈現明顯的細胞毒性（50%）。薑黃及蒼朮亦呈現細胞毒性，但是甘草之細胞毒性較小。如意金黃膏及croton oil不增加血中之IL-4及TNF-α，顯示如意金黃膏並無顯著之致敏性。疼痛科患者貼用如意金黃膏可改善疼痛，顯示如意金黃膏具有消炎止痛之潛力。此作用可能與一氧化氮合成?及第二型環氧化?之抑制與部份細胞毒性有關。如意金黃膏中之甘草是最具關鍵之植物成員，其細胞毒性較小，但是與抑制iNOS及COX-2有關之消炎作用最強。

Ruyi-Jinhuang Gao (RJG) is an ointment prepared from Ruyi-Jinhuang San (RJS), a traditional formula of powder-type Chinese medicine, usually used in anti-inflammatory analgesic care in China. The present study is aimed to detect the anti-inflammatory signaling of RJG exposed by an experimental model of lipopolysaccharides (LPS)-induced pro-inflammatory expression of inducible nitric oxide synthase (iNOS) and prostaglandin endoperoxide synthase (PGH synthase-2, COX-2). LPS-induced activation of iNOS and COX-2 has been recognized to increase cytokines and nitric oxide (NO), which play predominant roles in inflammation. In the culture of RAW264.7 cells, RJG concentration-dependently inhibited LPS-induced iNOS and COX-2 expression. However, the herbal components of RJG displayed different results, including increase and decrease of both protein expressions. Among them, inhibition by Curcuma Zedorarica Rhizoma (Cu), Atractylodis Lancea Rhizoma (At), and Glycyrrhizae Radix (Gl) are more potent than that by others. Inhibition by Cu and At are associated with their cytotoxicity. Glycyrrhizae Radix (Gl), with lower cytotoxicity in comparison with Cu and At, is the most potent component of RJG on inhibiting LPS-induced iNOS and COX-2 expression. In comparison with LPS-induced tumor necrosis factor-α (TNF-α), croton oil-induced formation of TNF-α and interleukin-4 (IL- 4), RJG did not increase the pro-inflammatory cytokines. These facts indicated that RJG could not significantly sensitize an immunologic response during the treatment of non-wounded inflammation on body surface. Ruyi-Jinhuang Patch (RJP), a pharmaceutical preparation of RJG, has proven to have significant relief of cutaneous inflammation-associated somatic pain in clinical use.