黄芪蜜炙对RAW264.7细胞能量代谢及极化的影响  

Effects of honey-processed Astragalus on energy metabolism and polarization of RAW264.7 cells

作　　者：李鸿昌 裴科 解旺洋 孟祥龙 于子涵 李玟玲 蔡皓[2] LI Hong-chang;PEI Ke;XIE Wang-yang;MENG Xiang-long;YU Zi-han;LI Wen-ling;CAI Hao(School of Chinese Medicine and Food Engineering,Shanxi Key Laboratory for Chinese Medicine Processing,Shanxi University of Chinese Medicine,Jinzhong 030619,China;School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]山西中医药大学中药与食品工程学院,中药炮制山西省重点实验室,山西晋中030619 [2]南京中医药大学药学院,江苏南京210023

出　　处：《药学学报》2025年第2期459-470,共12页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(82074022);山西省科技创新人才团队项目(202304051001044);国家自然科学基金联合基金重点项目(U21A20410);分子中医药学国际联合研究中心开放课题基金资助项目(2022JD-KF-01);山西省中医药管理局项目(2023ZYYA033);山西中医药大学科研项目(2022PY-TH-13)。

摘　　要：本研究以RAW264.7细胞为研究对象,观察黄芪蜜炙对其能量代谢及极化的影响,探讨蜜炙增强黄芪补中益气功效的科学内涵。将生品黄芪及蜜炙黄芪水煎液灌胃大鼠制备含药血清,采用液质联用技术分析黄芪蜜炙前后含药血清中成分的变化。通过CCK-8实验检测细胞存活率,筛选含药血清浓度。通过中性红吞噬实验、ATP检测试剂盒和NO检测试剂盒评价生品黄芪及蜜炙黄芪对细胞吞噬率、ATP能量代谢和NO分泌量的差异。通过ELISA试剂盒检测生品黄芪及蜜炙黄芪对RAW264.7细胞TNF-α分泌量的影响。通过流式细胞术评价生品黄芪及蜜炙黄芪对RAW264.7细胞极化的影响。采用液质联用技术筛选细胞裂解液和培养液中与糖酵解相关的差异代谢物。本实验获得山西中医药大学实验动物伦理委员会批准(批准号:AWE202407352)。结果显示,黄芪蜜炙后的水煎液灌胃大鼠后制备的含药血清中甜菜碱、氨基酸和芒柄花苷等成分含量升高。CCK-8实验结果表明,生品黄芪和蜜炙黄芪各浓度含药血清对RAW264.7细胞均无细胞毒作用。含药血清给药细胞后,吞噬指数和ATP的产量均有不同程度的提高,正常状态下RAW264.7细胞的NO分泌量提高,炎症状态下RAW264.7细胞的NO分泌量减少,且蜜炙黄芪的效果优于生品黄芪。ELISA试剂盒的检测结果表明,生品黄芪及蜜炙黄芪的含药血清均能呈浓度依赖性地促进TNF-α的分泌,且蜜炙黄芪的促进效果强于生品黄芪。流式细胞术的检测结果表明,生品黄芪及蜜炙黄芪的含药血清均能促进RAW264.7细胞向M1型极化,并抑制向M2型极化,蜜炙黄芪的效果优于生品黄芪。与生品黄芪相比,蜜炙黄芪的含药血清细胞裂解液和培养液中与糖酵解相关的代谢物总体呈上升趋势,表明蜜炙黄芪促进糖酵解的效果优于生品黄芪。综上所述,蜜炙黄芪能促进RAW264.7细胞的极化及能量代谢,并参与正向的�In this study,RAW264.7 cells were employed to investigate the effects of honey-processed Astragalus on their energy metabolism and polarization,and explore the scientific connotation of the enhanced efficacy of honey-processed Astragalus on invigorating spleen-stomach and replenishing Qi.The medicated sera were prepared by intragastric administration of rats with water extracts of crude and honey-processed Astragalus,and the composition changes of medicated sera of crude and honey-processed Astragalus were analyzed by using LC-MS technology.The cell survival rates were detected and the concentrations of medicated sera were screened through CCK-8 assay.The differences of cell phagocytic rates,ATP energy metabolism,and NO secretion between crude and honey-processed Astragalus were evaluated by using neutral red phagocytosis assay,ATP detection kit,and NO detection kit.The effects of crude and honey-processed Astragalus on TNF-α secretion of RAW264.7 cells were detected by employing ELISA kit.The effects of crude and honey-processed Astragalus on polarization of RAW264.7 cells were evaluated by utilizing flow cytometry.The differential metabolites related to glycolysis in cell lysates and culture media were screened by using LC-MS technology.The experiment was approved by the experimental animal ethics committee from Shanxi University of Chinese Medicine(No.AWE202407352).The results showed that the contents of betaine,amino acids,and ononin in the prepared medicated sera of rats treated by intragastric administration with water extracts of honey-processed Astragalus increased compared to those in crude one.The results of CCK-8 experiment showed that there were no cytotoxic effects on RAW264.7 cells in the medicated sera of crude and honey-processd Astragalus at different concentration.The phagocytic index and ATP yield both increased to varying degrees after administration of the medicated sera to cells.The secretion of NO in normal and inflammatory cells increased and decreased respectively,and the effect of honey

关 键 词：生品和蜜炙黄芪 能量代谢 极化 补中益气 

分 类 号：R917[医药卫生—药物分析学]