非類固醇抗炎藥與天然物合併治療對抗發炎反應之協同作用

Synergistic Effects of NSAIDs with Natural Products on Inflammatory Responses

曾頌惠(Sung-Hui Tseng)

指導教授 ： 楊玲玲 ； 王靜瓊

臺北醫學大學/藥學院/生藥學研究所/碩士(2002年)

摘要

中文摘要 發炎時所產生的紅、腫、熱、痛的生理現象，從分子生物科學探討，事實上包含了許多細胞內相關路徑的啟動，與細胞介質的相互調控，以期讓生物體能盡快回復到生理平衡狀態。臨床上，非類固醇消炎藥為治療發炎最常用的藥物，但往往只能發揮控制症狀的療效，因而很多台灣的民眾中西藥合併服用以加強治療效果。因此了解中西藥合併治療時是否會產生加乘或拮抗的作用，是一項重要的課題。故本研究從從中藥基準方中篩選出二十三種常用於治療關節炎之中藥材進行其抗發炎研究並探討其與西藥之協同作用。杜仲、黃芩、黃柏50%酒精萃取物對以lipopolysaccharide (LPS)誘導的老鼠巨噬細胞RAW 264.7所產生的Nitric Oxide (NO)有明顯的抑制作用。獨活、杜仲、紅花、黃芩、黃柏、大黃、麻黃、附子、當歸對prostaglandin E2 (PGE2) 有明顯的抑制作用。當萃取物與indomethacin同時處理LPS誘導後之細胞後，大部分的萃取物對NO的抑制作用有明顯的增加；但大黃、黃芩、黃柏，萃取物與celebrex同時處理時，只有續斷對NO的產量有明顯的增加抑制作用。另外，再以carrageenin誘導大鼠胸肋膜炎肢體內抗發炎模式，進一步了解中西藥合併使用對抗發炎作用是否有影響，結果顯示：單獨使用烏藥與烏藥加indomethacin組效果優於indomethacin或NO抑制劑(L-NAME)單獨使用。由上述結果得知，中西藥合併治療並非完全不可行。 繼而針對白朮、大黃、黃芩中具有抗發炎效果的指標成分atractylenolide I, rhein, emodin, wogonin, oroxylin A, baicalein, baicalin 與非類固醇消炎藥(indomethacin, celebrex,) 合併處理LPS活化的RAW 264.7 細胞，來探討其對NO 與PGE2加成或拮抗的結果及機轉。結果顯示二種非類固醇消炎藥皆能明顯增加atractylenolide I抑制NO的效果，而atractylenolide I亦不影響二種非類固醇消炎藥對PGE2的抑制率。但只有indomethacin能加乘emodin與 rhein對NO的抑制率，同時indomethacin對PGE2的抑制率亦不會受到影響。Wogonin, oroxylin A, baicalein, baicalin 對NO的抑制率在加入NSAIDs 後也沒有明顯的增加。因而推測：一，非選擇性的NSAIDs對天然物之抗發炎效果協同作用較強；二，黃芩與大黃之抗發炎活性成分與indomethacin合併並無增強NO的抑制作用，而致萃取物一無協同作用；三，用中藥中已知之天然物合併西藥，探討中西藥合併之作用，間接可探討中西藥合併是否具可行性為重要之實驗模式。

關鍵字

誘導型一氧化氮酵素 ； 環氧化酵素 ； 抗發炎 ； 老鼠巨噬細胞 ； 胸肋膜炎 ； 天然物 ； 非類固醇抗炎藥

並列摘要

Combine therapy of different classes of drugs in treatment of those diseases which are difficult to gain good clinical result, such as cancer, hypertension, rheumatoid arthritis, has gained more popularity due to better result and less side effects in the past decade. In the field of inflammation, many natural compounds have been found to possess anti-inflammatory property. Therefore, we investigated the effects of Chinese herbs in the absence or presence of NSAIDs on antiinflammatory responses. We first sorted out 23 most frequently used herbs for treating arthritis. These 23 herbs were than extracted with 50 % ethanol. The anti-inflammatory effects of these extracts were examined on LPS-induced NO and PGE2 production and expression of iNOS and COX-2 proteins in RAW 264.7 murine macrophage. Eucommia Cortex, Scrutelleriae Radix, and Phellodendri Cortex showed significant inhibition on NO production by LPS-activated Raw 264.7 cells. Angelicae Laxiflorae Radix, Carthami Flos, Angelicae Sinensis Radix, Aconiti Tuber, Rhei Rhizoma, Eucommia Cortex, Ephedra Herba, Scrutellariae Radix, and Phellodendri Cortex, 9 out of the 23 crude extracts, showed significant inhibition on PGE2 production . Combined treatment of the 23 herbs with indomethacin increased 15 herbs’ inhibition on NO production . However, when celebrex ( COX-2 inhibitor) was used to combined with the 23 herbs, the inhibitory effect of most of the herbs on NO production by LPS-activated RAW 264.7 cells were antagonized. We then determined whether combined treatment in vivo produces better anti-inflammatory effect. Carrageenin induced pleurisy in rats is a well established model of acute inflammation, and NSAIDs have been reported to inhibit a variety of inflammatory parameter. Adult male Wistar rats were used in the experiment. Drugs including NO inhibitor(L-NAME), indomethacin, or both, crude extract of Linderae Radix or with indomethacin were given 30min i.p. before the induction of pleurisy by 1% carrageenin. The group of rat treated with Linderae Radix or Linderae Radix with indomethacin showed less exudates than the rats treated with indomethacin or L-NAME. In the second part , we investigated the effects of anti-inflammatory compounds from Atractylodes Rhizoma(atractylenolide I), Scrutellariae Radix(wogonin, baicalein, baicalin, oroxylin A) and Rhei Rhizoma (emdin, rhein)on LPS-stimulated NO and PGE2 production in RAW 264.7 cells. The inhibitory effects when the compounds were combined with COX inhibitors were also investigated. The results indicated that both COX inhibitors could increase the inhibitory effect of atractylenolide I on NO production by LPS-induced RAW 264.7 cells. Only indomethacin could enhance the inhibitory effect of emodin and rhein on NO production by LPS-induced RAW 264.7 cells. The inhibitory effects on NO production of wogonin, baicalein, and oroxylin A did not increase after combined with NSAIDs. From the above results, we thus suggested the following: 1, Nonselective COX inhibitors can enhance the anti-inflammatory effect of natural compounds more significantly than selective COX inhibitors; 2, NSAIDs could antagonized the anti-inflammatory effect of Scrutellariae Radix and Rhei Rhizoma and was most likely due to NSAIDs could antagonize the anti-inflammatory effect of wogonin, baicalein, and oroxylin A; 3, the models used in the experiments could provide useful informations for future research on the combined treatment of medicine and Chinese herbs.

並列關鍵字

inducible nitric oxide ； cyclooxygenase ； antiinflammation ； RAW264.7 ； pleurisy ； natural compounds ； NSAIDs

參考文獻

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