黄精对白细胞介素-4诱导M2巨噬细胞能量代谢和极化的作用与机制  

Effects and mechanism of Polygonati Rhizoma on energy metabolism and polarization of interleukin-4-induced M2 macrophages

作　　者：侯亚琴 李明[2] 岳利多 郝冰洁 岳庆喜 范理宏 HOU Ya-qin;LI Ming;YUE Li-duo;HAO Bing-jie;YUE Qing-xi;FAN Li-hong(Shanghai Clinical College,Anhui Medical University,Hefei 230032,China;Shanghai Tenth People’s Hospital Affiliated to Tongji University,Shanghai 200072,China;Institute of Energy Metabolism and Health,School of Medicine,Tongji University,Shanghai 200072,China)

机构地区：[1]安徽医科大学上海临床学院,合肥230032 [2]同济大学附属第十人民医院,上海200072 [3]同济大学医学院能量代谢与健康研究所,上海200072

出　　处：《中华中医药杂志》2022年第8期4400-4404,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.31770131);上海市科学技术委员会一带一路联合实验室项目(No.20400750600)。

摘　　要：目的:探讨黄精对M2巨噬细胞能量代谢和极化的影响及其分子机制。方法:用白细胞介素-4(IL-4)构建M2巨噬细胞模型;CCK-8细胞增殖实验检测细胞活力;qPCR和Western blot实验检测M2巨噬细胞标志Arg1、CD206mRNA及蛋白和AMPK/PDH信号通路(p-AMPK、p-PDH、PDK)蛋白的表达;细胞能量代谢分析仪(Seahorse)分析M2巨噬细胞能量代谢;ATP检测试剂检测M2巨噬细胞总ATP产量。结果:用IL-4处理RAW264.7巨噬细胞和THP-1细胞成功构建M2巨噬细胞模型。黄精浓度<2.5 mg/mL时对巨噬细胞无明显细胞毒作用。黄精可抑制M2巨噬细胞标志Arg1和CD206mRNA及蛋白的表达(P<0.01,P<0.05),且抑制程度呈浓度依赖性;黄精可以降低M2巨噬细胞ATP产量、耗氧率(P<0.05,P<0.01),提升细胞外酸化率(P<0.01);黄精抑制p-AMPK、p-PDH蛋白的表达(P<0.05,P<0.01),黄精联合AMPK抑制剂使用后,p-AMPK、p-PDH蛋白表达更低(P<0.05)。结论:黄精抑制M2巨噬细胞线粒体氧化磷酸化,降低M2巨噬细胞ATP产量,抑制M2巨噬细胞极化,其机制与黄精抑制AMPK/PDH信号通路磷酸化相关。Objective:To explore the effects of Polygonati Rhizoma on energy metabolism and polarization of M2macrophages and its molecular mechanism.Methods:The M2 macrophage model was induced with interleukin-4(IL-4);CCK-8experiment detected cell vitality;qPCR and Western blot experiments were used to detect the mRNA and protein of Arg-1and the expression of CD206 and AMPK/PDH signaling pathway related proteins(p-AMPK,p-PDH,PDK);Cell Energy Metabolism Analyzer(Seahorse) analyzed the energy metabolism of M2 macrophages;enhanced ATP detection reagent detects the ATP production of M2 macrophages.Results:The M2 macrophage model was successfully constructed by treating RAW264.7macrophages and THP-1 cells with IL-4.There was no obvious cytotoxic effect on macrophages when the Polygonati Rhizoma concentration<2.5 mg/mL;Polygonati Rhizoma inhibited the expressions of M2 macrophage marker Arg1 and CD206 mRNA and protein(P<0.01,P<0.05),and its inhibitory effect was in a concentration-dependent manner;Polygonati Rhizoma reduced the ATP production,oxygen consumption rate of M2 macrophages(P<0.05,P<0.01),and improved the extracellular acidification rate(P<0.01).Polygonati Rhizoma inhibited the expression of p-AMPK and p-PDH proteins(P<0.05,P<0.01),and the expression of p-AMPK and p-PDH protein was lower when Polygonati Rhizoma combined with AMPK inhibitor(P<0.05).Conclusion:Polygonati Rhizoma down-regulates mitochondrial oxidative phosphorylation,reduces the ATP production in M2 macrophages,and inhibits M2 macrophage polarization and its mechanism maybe related to the inhibition of AMPK/PDH signaling pathway phosphorylation by Polygonati Rhizoma.

关 键 词：黄精 M2巨噬细胞 能量代谢 AMPK/PDH信号通路 肿瘤免疫 极化 

分 类 号：R285[医药卫生—中药学]