Auricularia auricula-judae Hot Water Extracts Activate Macrophages Toward M1 Phenotype

黑木耳熱水萃取物活化巨噬細胞趨向M1功能型

鍾惠貞(Hui-Chen Chung) ； 余哲仁(Zer-Ran Yu) ； 王璧娟(Be-Jen Wang) ； 林淑美(Shu-Mei Lin)

《臺灣農業化學與食品科學》 57卷5&6期 (2019/12) Pp. 220-232

https://doi.org/10.6578/TJACFS.201912_57(5_6).0003

摘要

Proper modulation of immune response is a novel strategy for disease prevention and clinical treatment. Edible mushrooms are potent dietary sources of natural immune modulators which components are highly diversified in biological function, chemical structure and molecular weight. In the present study, we determined the immune-modulating activity of the crude hot water extract of Auricularia auricula-judae (AACE). AACE was fractionized into four subfractions based on molecular weight, assigned as F1 (＜10 kDa), F2 (10-50 kDa), F3 (50-100 kDa), and F4 (＞100 kDa). The immune-modulating activity of AACE and the sub-fractions was evaluated in murine macrophages. The results indicated that crude hot water extract and the subfractions, except low molecular weight-fraction (F1), exhibited immune-stimulating, rather than anti-inflammatory activity, by triggering macrophage activation toward M1-like functional phenotype determined by the secretion of inflammatory immune mediators, including NO, TNF-α, IL-1, and IL-6, and the induction of M1 cell specific cell surface marker, CD11c. F2 and F4 exhibited the highest capacity of macrophage activation among AACE and the sub-fractions. At the concentration of 25 μg/mL, F2 and F4 fractions exhibited similar potency of macrophage-stimulating activity as 0.5 μg/mL lipopolysaccharide, while F1 (＜10 kDa) showed limited activating effect even at the dosage of 100 μg/mL AACEs stimulated macrophages likely through toll-like receptor-4 (TLR-4) signaling. Supplementation of AACE, F2 or F3, at 50 μg/mL, exerted anti-proliferative effect on cancer cells co-cultured with macrophages. These results suggested that A. auricula-judae was a dietary source of potent immunostimulants comprising various types of macrophage-activating components. The most potent immunostimulants were the components with molecular weight at 10-50 kDa or higher than 100 kDa, and elimination of low molecular weight (＜ 10 kDa) components might enhance AACE immune-stimulating activity. Our study provides scientific data supporting nutraceuticals application of A. auricula-judae and might prompt further studies to identify specific immunostimulant compounds in AACE for nutraceutical application.

關鍵字

Auricularia auricula-judae ； Dietary adjuvant ； Immunostimulant ； Macrophage activation ； M1 phenotype ； Immunotherapy

並列摘要

免疫系統的調節是疾病預防治療的重要策略。食用真菌具多種生物活性，更是免疫調節功能化合物的重要膳食來源。各類真菌所含的功能性成分在生物功能、化學結構及分子大小上極具多樣化。本研究主要探討常用食用真菌－黑木耳（Auricularia auricula-judae）的免疫調節功效。黑木耳熱水粗萃物（AACE）依分子量區分為F1（＜10 kDa）、F2（10-50 kDa）、F3（50-100 kDa）及F4（＞100 kDa）。並以小鼠巨噬細胞株（Raw264.7）作為試驗模式。結果顯示，除低分子量區分物（F1）外，黑木耳粗萃物和其他區分物均可誘導巨噬細胞活化並使其趨向M1功能表現型，包括誘發發炎反應調節因子－NO，以及TNF-α、IL-1和IL-6等細胞激素的釋放，並促進M1細胞特異基因－CD11c的表現。黑木耳粗萃物及所有區分物相較，以F2及F4活化巨噬細胞的能力最強。25 μg/mL的F2與F4活化巨噬細胞之效能相當於0.5 μg/mL脂多醣（LPS）的誘發活性。F2、F4及AACE活化巨噬細胞主要是透過TLR-4途徑。而黑木耳萃取物刺激巨噬細胞活化與其抗癌效能有關。在肺癌細胞及巨噬細胞共培養下，補充50 μg/mL的AACE、F2或F3區分物具有抑制癌細胞增殖的效能。以上證實黑木耳含有多種活化巨噬細胞之功能性成分，具有活化免疫反應的功效，可作為免疫調節相關保健食品或特殊疾病營養配方（例如癌症免疫療法膳食輔佐劑）的開發應用。

並列關鍵字

黑木耳 ； 膳食輔佐劑 ； 免疫刺激物 ； 巨噬細胞活化 ； M1功能型 ； 免疫治療