Nagai Kenta, Ueno Yoshitaka, Tanaka Shinji, Hayashi Ryohei, Shinagawa Kei, Chayama Kazuaki
Department of Medicine and Molecular Science, Hiroshima University, Japan.
Department of Endoscopy, Hiroshima University, Japan.
Cell Immunol. 2017 Oct;320:20-28. doi: 10.1016/j.cellimm.2017.08.001. Epub 2017 Aug 4.
Non-steroidal anti-inflammatory drugs often cause ulcers in the human small intestine, but few effective agents exist to treat such injury. Ganoderma lucidum Karst, also known as "Reishi" or "Lingzhi", is a mushroom. We previously reported that a water-soluble extract from G. lucidum fungus mycelia (MAK) has anti-inflammatory effects in murine colitis induced by trinitrobenzene sulfonic acid, and induction of granulocyte macrophage colony-stimulating factor (GM-CSF) by MAK may provide anti-inflammatory effects. However, its effects on indomethacin-induced small intestinal injuries are unknown. The present study investigated the preventative effects of MAK via immunological function and the polysaccharides from MAK on indomethacin-induced ileitis in mice. Peritoneal macrophages (PMs) were stimulated in vitro with MAK and adoptively transferred to C57BL/6 mice intraperitoneally, which were then given indomethacin. Intestinal inflammation was evaluated after 24h. We performed in vivo antibody blockade to investigate the preventive role of GM-CSF, which derived from PMs stimulated with MAK. We then used PMs stimulated with MAK pre-treated by pectinase in an adoptive transfer assay to determine the preventive role of polysaccharides. Indomethacin-induced small intestinal injury was inhibited by adoptive transfer of PMs stimulated in vitro with MAK. In this transfer model, pre-treatment with anti-GM-CSF antibody but not with control antibody reversed the improvement of small intestinal inflammation by indomethacin. Pectinase pretreatment impaired the anti-inflammatory effect of MAK. PMs stimulated by MAK appear to contribute to the anti-inflammatory response through GM-CSF in small intestinal injury induced by indomethacin. The polysaccharides may be the components that elicit the anti-inflammatory effect.
非甾体抗炎药常导致人类小肠溃疡，但治疗此类损伤的有效药物很少。灵芝，也被称为“赤芝”或“灵芝”，是一种蘑菇。我们之前报道过，灵芝真菌菌丝体的水溶性提取物（MAK）在三硝基苯磺酸诱导的小鼠结肠炎中具有抗炎作用，并且MAK诱导粒细胞巨噬细胞集落刺激因子（GM-CSF）可能发挥抗炎作用。然而，其对吲哚美辛诱导的小肠损伤的影响尚不清楚。本研究通过免疫功能和MAK中的多糖来研究MAK对吲哚美辛诱导的小鼠回肠炎的预防作用。用MAK体外刺激腹腔巨噬细胞（PMs），然后将其腹腔内过继转移到C57BL/6小鼠体内，随后给小鼠注射吲哚美辛。24小时后评估肠道炎症。我们进行了体内抗体阻断实验，以研究源自用MAK刺激的PMs的GM-CSF的预防作用。然后，我们在过继转移实验中使用经果胶酶预处理的用MAK刺激的PMs来确定多糖的预防作用。用MAK体外刺激的PMs过继转移可抑制吲哚美辛诱导的小肠损伤。在这个转移模型中，用抗GM-CSF抗体而非对照抗体预处理可逆转吲哚美辛对小肠炎症的改善作用。果胶酶预处理削弱了MAK的抗炎作用。MAK刺激的PMs似乎通过GM-CSF在吲哚美辛诱导的小肠损伤中发挥抗炎反应。多糖可能是发挥抗炎作用的成分。
