Cordyceps sinensis, a traditional medicinal fungus, has been widely used in China for its various biological activities. However, its usage has been limited during the past decades due to high price and rarity, so isolation of mycelial strains from Cordyceps to achieve a large-scale production by fermentation has become a trend. The aim of this study is to investigate the effects of water extracts of two natural Cordyceps, C. sinensis (DCXC) and C. gunni (GNCC) and fifteen cultured Cordyceps mycelia, which the fungus strains from natural C. sinensis (LZST4, QHIIJC, BNQ, BNZK, QHJ, 50542, 50562, 50677, HS-1, HS-2, WJM, PHUM, BMB and CNB) and C. militaris (BCC), on macrophage function in vitro. The assays included proliferation, production of NO, iNOS protein levels, phagocytosis, pro-inflammatory cytokines, and translocation of NF-kB and protein expression of NF-KB p65 on RAW 264.7 mouse macrophage cells. Cell viability, NO production, phagocytic activity, pro-inflammatory cytokines, NF-KB activation and expressions of iNOS and NF-KB p65 were investigated by MTT assay, colorimetric method, flow cytometry, ELISA, immunostaining and Western Blot, respetively. The results showed that two natural Cordyceps (DCXC and GNCC and three cultured Cordyceps mycelia (LZST4, QHI1JC and BNQ) significantly increased cell viability and production of NO, induced expression of iNOS and enhanced phagocytosis of RAW 264.7 macrophage cells. DCXC, GNCC, LZST4, QH11JC and BNQ significantly increased the secretion of TNE-o. High concentration of LZST4 and QH11JC, moreover, promoted the production of IL-1a, IL-1ß and IL-10. DCXC at high concentration also increased the secretion of IL-1ß and IL-10, and GNCC increased the production of IL-10. In addition, LZST4 markedly induced the translocation of NF-KB in macrophages and increased p-NF-KB p65 expression on RAW 264.7 macrophages. Therefore, natural C. gunni (GNCC) and cultured Cordyceps mycelia, LZST4, QH11JC and BNQ, have similar effects with natural C. sinensis (DCXC) on RAW 264.7 macrophages.

冬盐豆草是中國傅統的名武滋補中藥材，與人參，鹿茸並列為三大滋神品，員有多種藥理活性。逦去雞十年，由於價格昂費和资源稀缺導致夭総蟲章的使 用受到極大限制。因此從天然冬蟲豆草中分離菌株，並投入到大規模生產成為 蟲草產業發展的必然趨勢。 本研究的目的旨在考察兩種天然蟲草，冬蟲夏草 (DCXC）和古尼蟲草 (GNCC) 及十五種人工蟲草菌絲體，其中14 種為從天然冬蟲复草中分離所得 ## (LZST4, QHI1JC, BQ, BNZK, QHJ. 50542, 50562, 50677, HS-1, HIS-2、WIM、FEIM、BMB 和 CNB）和1種為北蟲草(BCC)菌株，水提物對 RAW 264.7 巨噬細胞功能的影響，比較天然和人工蟲草免疫作用的差異。实验内容包括蟲草水提物對 RAW 264.7 巨噬細胞的細胞增殖、吞噬功能、NO释放、iNos 蛋白表达、細胞因子釋放、NF-KB 核轉位和 NF-KB p65 蛋白表达的影響。 實驗採用 MTT 法檢測細胞增殖作用、Griess 試劑檢測 NO 的釋放量、流式細 胞檢測儀檢測細胞的吞噬能办 ELISA 法定量細胞因子 螢光標記法觀察 NF-KB 的核轉位和 Western Blot 法检测 iNOS 及NF-KB p65 的蛋白表達。 實驗結果表明兩種天然蟲草(DCXC 和 GNCC) 和三種人工蟲草菌絲體 (LZST4、QH11JC 和BNQ）可顯著增强 RAW 264.7 巨噬細胞活性，增加 NO 释放、iOS 蛋白表達和提高巨噬細胞吞噬能力。另外，这五種蟲草可以不同 程度提高巨噬細胞細胞因子 TNF-Q的釋放，LZST4 和 QH11JC 在高濃度時還可 以增加 兀-10 IL-18 和I-10的釋放，DCXC 在高濃度時可增加I-18 和IL-10 的释放，GNCC 也能促进I-10的釋放。Western Blot 實驗結果表明 只有LZST4 能增加磷酸化 NF-KB p65 蛋白表達，免疫荧光标记试验也进一步证实 LZST4 可诱导 NF-KB 核转位。 本研究显示 GNCC、LZST4、QH11JC 和 BNQ 與天然冬蟲夏草 DCXC具 有相似的顯著增强 RAW 264.7 巨噬細胞功能的作用，值得進一步研究。