銀杏萃取物與柴胡皂苷 a 對脂多醣誘導人類單核球 U-937 細胞 
發炎物質之影響

發炎反應為清除入侵體內病原而產生的防禦機制，而持續或過度的發炎反應則會對身體組織造成傷害，進而引發許多慢性疾病。銀杏萃取物 (EGb 761) 中有大量類黃酮與其特有之銀杏內酯與白果內酯，已有許多研究證實其具有抗氧化、改善血流、改善記憶等效用，而 EGb 761 對於抗發炎之效用其機制卻尚未明瞭。柴胡複方常被作為治療肝炎用藥，柴胡皂苷 a (saikosaponin a, SSa) 為其主要功效成分之一，研究發現其可能具有抗發炎之效用，但相關文獻仍非常稀少。本次實驗探討 EGb 761 與 SSa 對於以脂多醣 (lipopolysacchrides, LPS) 刺激分化後之人類單核球 U-937 細胞所產生發炎物質之影響。將細胞培養於含10% 胎牛血清之 RPMI 1640 培養液中，並置於 37℃、含 5% CO2 的細胞培養箱，以 160 nM phorbol 12-myristate 13-acetate 培養 48 小時誘導細胞分化為巨噬細胞，之後更換為無血清之 0.1% 牛血清蛋白培養液繼續培養。將細胞培養於含 200 μg/ml EGb 761、5 μg/ml SSa 或 200 μg/ml EGb 761 + 5 μg/ml SSa 之培養液 24 小時，再更換為含 1 μg/ml LPS 之培養液培養 24 小時後，收集培養液與細胞液進行發炎相關物質之分析。結果顯示 EGb 761、SSa 及 EGb 761 + SSa 組皆顯著降低培養液中因 LPS 刺激而釋放之腫瘤壞死因子-α (tumor necrosis factor-α, TNF-α)、介白素-1β (interleukin-1β)、介白素-10 (interleukin-10)、前列腺素 E2 (prostaglandin E2) 濃度及細胞中第二型環氧化酵素 (cyclooxygenase-2) 蛋白質表現量 (p<0.05)。此外也發現 200 μg/ml EGb 761 可清除 sodium nitroprusside 產生之一氧化氮 (nitric oxide, NO) (p<0.05)。經加乘效果公式計算後，發現 EGb 761 與 SSa 對刺激 TNF-α 分泌量與 NO 清除能力具有加乘效果。因此本實驗結果發現 200 μg/ml EGb 761 與 5 μg/ml SSa 能夠降低 LPS 刺激 U-937 巨噬細胞刺激而產生之發炎物質與清除 NO，並具加乘效果，因而可能具有抑制發炎之效用。

關鍵字

EGb 761 ；柴胡皂苷a ； U-937 ；發炎

並列摘要

Inflammation is a defense mechanism to scavenge the invaded pathogens. Sustained or excessive inflammation would cause harmful to the tissues, and further trigger many chronic diseases. Ginkgo biloba extract (EGb 761) contains a large number of flavonoids, ginkgolides and bilobalide, and many studies have reported its activities on enhancing antioxidative defense, increasing blood flow, and improving memory. However, the anti-inflammatory effect of EGb761 has not been clear. Bupleurum is often used as a compound for the treatment of hepatitis. Saikosaponin a (SSa), Bupleurum's major active ingredient, which was found to have anti-inflammatory effect, but very few studies have reported. This study investigated the effects of EGb 761 and/or SSa on lipopolysaccharide (LPS)-induced inflammatory responses in differentiated human monocytes U-937. Cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS), and placed in 37℃ and 5% CO2 incubator. The differentiation from monocytes to macrophages was induced 160 nM phorbol 12-myristate 13-acetate for 48 hours. After differentiation, the medium was replaced by FBS-free medium containing 0.1% bovine serum albumin. Cells were then cultured with 200 μg/ml EGb 761, 5 μg/ml SSa, or 200 μg/ml EGb 761 + 5 μg/ml SSa for 24 hours, and 1 μg/ml LPS was added to the medium for another 24 hours. Finally, the conditioned medium and cell lysate were collected. The results showed that 200 μg/ml EGb 761, 5 μg/ml SSa and 200 μg/ml EGb 761 + 5 μg/ml SSa treatments significantly decreased the secretion of LPS-induced tumor necrosis factor-α (TNF-α), interleukin-1β, interleukin-10 and prostaglandin E2 and protein expression of cyclooxygenase-2 (p<0.05). Additionally, 200 μg/ml EGb 761 could scavenge nitric oxide (NO) which produced by sodium nitroprusside (p<0.05). Finally, we found EGb 761 and SSa have synergistic effects on TNF-α seceretion and NO scavenging ability. Therefore, 200 μg/ml EGb 761 and 5 μg/ml SSa alleviate inflammation through decreasing inflammatory substances produced by LPS-induced U-937 macrophages, and scavenging NO. Furthermore, EGb 761 and SSa have synergistic effects.

並列關鍵字

EGb 761 ； saikosaponin a ； U-937 ； inflammation

參考文獻

洪慧珍 (2002) 銀杏葉萃取液對十二指腸潰瘍老鼠黏膜修復之影響。臺北醫學大學保健營養學研究所碩士論文。

Ahmad N, Srivatava RC, Agarwal R, Mukhtar H (1997) Nitric oxide synthase and skin tumor promotion. Biochem Biophys Res Commun 232: 328-331.

Alexander C, Rietschel ET (2001) Bacterial lipopolysaccharides and innate immunity. J Endotoxin Res 7: 167-202.

Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J, Pinsky MR (2001) Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care. Critical Care Medicine 29: 1303-1310.

Arend WP (1991) Interleukin-1 receptor antagonist. A new member of the interleukin-1 family. J Clin Inves 88: 1445-1451.

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銀杏萃取物與柴胡皂苷 a 對脂多醣誘導人類單核球 U-937 細胞發炎物質之影響

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