冠突散囊菌与木霉混合发酵液醇提物对小鼠巨噬细胞RAW264.7免疫调节的影响

为进一步开发冠突散囊菌的药理活性，该研究将冠突散囊菌与木霉共同发酵，通过不同体积分数的乙醇对混合发酵液化学活性部位进行追踪，并结合液质联用（LC-MS）技术对活性部位化合物组成进行分析，通过四氮甲基唑蓝（MTT）比色法、中性红吞噬实验及酶联免疫吸附测定方法分别研究冠突散囊菌与木霉混合发酵液活性部位对小鼠巨噬细胞RAW264.7增殖、吞噬以及分泌细胞因子的能力的影响。结果表明，冠突散囊菌与木霉混合发酵液体积分数70%乙醇提取部位有较好的免疫活性，该部位含有大黄素、芥子碱等生物活性成分。与阳性对照组脂多糖相比，500 μg/mL冠突散囊菌与木霉混合发酵液的体积分数70%乙醇提物能够使小鼠巨噬细胞RAW264.7增殖率提高12%，增强小鼠巨噬细胞RAW264.7吞噬中性红的能力，并能促进NO及细胞因子IL-6、TNF-α的分泌。

Effect of ethanol extracts from co-fermentation liquid of Eurotiunm cristatum and Trichoderma sp. on immunoregulation of mouse macrophages RAW264.7

To further excavate the pharmacological activity of Eurotiunm cristatum, E. cristatum and Trichoderma sp. were co-fermented. The chemical active parts of co-fermentation liquid were separated by different volume fractions of ethanol, and detected by LC-MS technology. The effects of the active parts of co-fermentation liquid of E. cristatum and Trichoderma sp. on the proliferation, phagocytosis and cytokines secretion ability of mouse macrophage RAW264.7 were determined by methyl thiazolyl tetrazolium (MTT) assay, neutral red phagocytosis tests and enzyme-linked immunosorbent assay, respectively. The results showed that volume fraction 70% ethanol extraction of the co-fermentation liquid of E. cristatum and Trichoderma sp. had good immune activity, and this part contained biologically active ingredients such as emodin and sinapine. Compared with the positive control group lipopolysaccharide, the volume fraction 70% ethanol extraction of the co-fermentation liquid of E. cristatum and Trichoderma sp. at 500 μg/ml could significantly increase the proliferation rate of mouse macrophages RAW264.7 by 12%, enhance the neutral red phagocytosis of mouse macrophages RAW264.7, and promote the secretion of NO and cytokines IL-6 and TNF-α.