幾種食藥用菇抗氧化、抗發炎活性及其抑制泡沫細胞形成能力

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幾種食藥用菇抗氧化、抗發炎活性及其抑制泡沫細胞形成能力

Antioxidant and anti-inflammatory activities of several edible and medicinal mushrooms and their inhibitory ability on foam cell formation

李佑軒 , Masters　　Advisor：毛正倫　　

繁體中文

舞菇；粉紅玫瑰菇；鴻喜菇；香杉芝；抗氧化；抗發炎；泡沫細胞； Grifola frondosa ； Pleurotus salmoneostramineus ； Hysizigus marmoreus ； Antrodia salmonea ； antioxidation ； anti-inflammation ； foam cell

Abstract │ Reference (223) │ Times Cited (3) │ Altmetrics

Abstract 〈TOP〉

適當的發炎反應可保護人體健康，但發炎反應過量分泌的發炎調控因子，將對細胞或器官產生致病性的影響。ROS在生物體內，同時扮演促進氧化壓力和誘導免疫細胞分泌發炎調控因子的角色。動脈粥狀硬化發展的關鍵為泡沫細胞之生成，泡沫細胞對此病灶的致病性具有決定性之影響。食藥用菇富含多種生理活性成分，具多種生理活性，是開發保健食品的良好素材。本研究目的主要探討舞菇(Grifola frondosa)、粉紅玫瑰菇 (Pleurotus salmoneostramineus)、鴻喜菇 (Hypsizigus marmoreus) 子實體和香杉芝 (Antrodia salmonea) 菌絲體70%乙醇萃取物，對抗發炎、抗氧化以及抑制巨噬細胞衍生為泡沫細胞之能力。

分析樣品中生理活性物質得知，舞菇子實體 (GF)、粉紅玫瑰菇子實體 (PS)、鴻喜菇子實體 (HM) 及香杉芝菌絲體 (AS) 之70%乙醇萃取物，總酚含量為 9.90-37.78 GAE mg/g extract，類黃酮為含量 0.43-10.46 RE mg/g extract，總酚及類黃酮均以AS含量最高。試管抗氧化能力分析結果指出，還原力以AS最佳，EC50濃度為0.86 mg/mL;清除DPPH能力方面，所有萃取物之EC50濃度，均大於20 mg/mL;螯合亞鐵離子能力方面，以PS最佳，EC50濃度為0.53 mg/mL。

抗發炎性質評估方面，探討萃取物對LPS誘導巨噬細胞J774.A1 24小時分泌發炎調控因子之影響。結果顯示，抑制NO效果以AS (40 μg/mL) 最好，抑制率達29.82%;抑制TNF-α效果以GF (500 μg/mL) 最好，抑制率達37.83%; 抑制IL-1β效果，HM (750 μg/mL) 及 AS (40 μg/mL) 實驗結果相似，均有良好抑制效果，其分泌量均判定為未檢出;抑制IL-6效果以GF (500 μg/mL) 最好，抑制率達80.97%;促進抗發炎細胞激素IL-10分泌之影響，以PS (200 μg/mL) 最能顯著提升IL-10分泌量。以促發炎/抗發炎細胞激素比值 (TNF-α+IL-6+IL-1β)/IL-10觀察整體細胞激素分泌之變化，結果顯示，GF (500 μg/mL)、PS (200 μg/mL) 及HM (750 μg/mL) 均顯著下降 促發炎/抗發炎細胞激素比值，達到抗發炎之效果。AS (20-40 μg/mL) 顯著提升促發炎/抗發炎細胞激素比值，可能具促進發炎反應之傾向。

胞內抗氧化評估方面，結果顯示，LPS誘導3及6小時，以AS (40 μg/mL)抑制效果最好，ROS生成量分別由109.12%下降至79.80%以及由221.53%下降至169.94%。LPS誘導12及24小時，以PS (200 μg/mL) 抑制效果最好，ROS生成量分別由267.10%下降至172.66%以及由229.24%下降至150.99%。

抑制生成泡沫細胞評估方面，結果顯示，以100?g/mL之oxLDL培養J774.A1 48小時，胞內脂質堆積量顯著上升至105.30%，以此作為建立巨噬細胞衍生為泡沫細胞之實驗平台。油紅染色實驗結果顯示，GF、PS、HM 和AS皆顯著提升J774.A1吞噬oxLDL能力，分別上升至101.82、103.84、103.92及102.84%，促進泡沫細胞之生成。探討萃取物對oxLDL誘導J774.A1及其衍生之泡沫細胞引起的發炎反應之影響，結果顯示，oxLDL無法誘導J774.A1及其衍生之泡沫細胞發炎分泌TNF-α、IL-6及IL-1β; 其中在PS、HM及AS處理下，可顯著促進巨噬細胞分泌TNF-α。

綜合以上實驗結果，GF、PS、HM及AS呈現了不同程度之抗氧化及抗發炎效果，另一方面，四種食藥用菇萃取物可能無法藉由抑制泡沫細胞生成或調節發炎反應來達到改善動脈粥狀硬化之發展。

Parallel Abstract 〈TOP〉

Appropriate inflammatory responses might protect human body from diseases, but excessive inflammation may be harmful to cells and organs leading to many   chronic diseases. ROS plays an important role in oxidative stress and modulates the secretion of inflammatory mediators. The formation of foam cells plays an important role in the development of atherosclerosis process. Edible and medicinal mushrooms contain various kinds of bioactive compounds, presenting multiple bioactivities. Thus, mushrooms are good sources to be used for health foods. In this study, the effect of 70% ethanolic extracts from Grifola frondosa (GF), Pleurotus salmoneostramineus (PS), Hysizigus marmoreus (HM) fruiting body and Antrodia salmonea (AS) mycelium on oxidation, inflammation and foam cells formation was investigated.

The results showed that the content of total phenols and flavonoids in GF, PS, HM and AS were 9.90-37.78 GAE mg/g extract and 0.43-10.46 RE mg/g extract, respectively. Among these extracts, AS has the highest content of total phenols and flavonoids. The in vitro antioxidant properties of extracts were evaluated. AS had superior activities on reducing power, and the EC50 were 0.86 mg/mL; on DPPH scavenging ability, the EC50 of all extracts were higher than 20 mg/mL; on the chelating ferrous ions, PS had superior activities, and the IC50 were 0.53 mg/mL.

According to the results of anti-inflammation, 125 μg/mL AS had superior effect on reducing the content of NO, and the inhibition was 29.82%; 500 μg/mL GF had superior effects on reducing the content of TNF-α and IL-6, and the inhibition were 37.83% and 80.97%, respectively; 750 μg/mL HM and 40 μg/mL AS had similar effect on reducing IL-1β production, the content were below the limit of detection. 200 μg/mL PS could significantly increase IL-10 production. The pro-/anti-inflammatory cytokine ratio by (TNF-α+IL-6+IL-1β)/IL-10 was also evaluated, to clarify the profile of various cytokines. The results indicated that 500 μg/mL GF, 200 μg/mL PS and 750 μg/mL HM exhibited great anti-inflammatory activities by significantly decreasing the pro-/anti-inflammatory cytokine ratios. In contrast, AS (20-40 μg/mL) could significantly increase the pro-/anti-inflammatory cytokine ratios. Overall, GF, PS and HM might have good anti-inflammatory potential, while AS might has pro-inflammatory potential.

The effect of extracts on LPS-induced ROS production in J774.A1 cells for 3, 6, 12 and 24 h was investigated. According to the results , after short induced time of 3 and 6 h, AS showed great inhibition on ROS production; after long induced time of 12 and 24 h, PS showed great inhibition on ROS production.

The cell model of foam cell formation was established by 100 μg/ml oxLDL induced J774.A1 to become foam cells after 48 h. According to the results of Oil red O staining, GF, PS, HM and AS could significantly increase the ability of macrophage to uptake oxidized modified low density lipoprotein (oxLDL). The effect of the extracts on oxLDL-induced inflammatory response in J774.A1 cells for 48 h was also evaluated. According to the result, oxLDL-induced J774.A1 could not release TNF-α, IL-1β and IL-6, which indicated that the inflammatory response did not occure. Among the extracts, PS, HM and AS induce J774.A1 activate, resulting in secretion of TNF-α.

In conclusion, GF, PS, HM and AS exhibited multiple antioxidant and anti-inflammatory properties. In the other hand, these extracts might not have good potential in decreasing the development of atherosclerosis by inhibiting the formation of foam cells and declining inflammatory responses.

Reference ( 223 ) 〈TOP〉

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