异槲皮素通过激活YAP蛋白对巨噬细胞极化表型的影响  

Effects of isoquercetin on the phenotype of macrophage polarization through activation of YAP protein

作　　者：戴淑颖 刘怿 孙霖 张郜晨茜 舒琦瑾[2] DAI Shu-ying;LIU Yi;SUN Lin;ZHANG Gao-chen-xi;SHU Qi-jin(First Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou 310053,China;Department of Oncology,Zhejiang Provincial Hospital of Chinese Medicine,Hangzhou 310060,China)

机构地区：[1]浙江中医药大学第一临床医学院,杭州310053 [2]浙江省中医院肿瘤科,杭州310060

出　　处：《中华中医药杂志》2023年第5期2444-2449,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82274596,No.81873243)。

摘　　要：目的:探讨异槲皮素(I S O)对巨噬细胞极化表型的影响及相关分子机制。方法:体外培养RAW264.7、THP-1细胞株,采用CCK-8法检测ISO作用后巨噬细胞活力。根据CCK-8结果设置低、中、高剂量ISO组,采用Western Blot、RT-qPCR、流式细胞术检测ISO对M0及M2型巨噬细胞中M2型标志物及Yes相关蛋白(YAP)表达水平的影响。此外,加入YAP抑制剂,检测其对巨噬细胞极化表型的影响。结果:在M0及M2巨噬细胞模型中,ISO作用巨噬细胞24 h后,细胞中M2型标志物甘露糖受体(CD206)及YAP表达量增加(P<0.05,P<0.01)。流式细胞术也证实ISO作用后巨噬细胞中CD206平均荧光强度(MFI)显著升高(P<0.05,P<0.01)。此外,YAP抑制剂能减弱ISO上调巨噬细胞CD206表达(P<0.01)。结论:异槲皮素作为免疫调节剂,可以通过激活YAP从而促进巨噬细胞抗炎M2表型的极化。Objective:To explore the effects of isoquercetin(ISO)on the polarization of macrophages and the underlying mechanisms.Methods:The RAW264.7 and THP-1 cell lines were cultured in vitro,while the cell viability after ISO treatment was detected by the cell counting Kit-8(CCK-8)method.According to the results of CCK-8,low,medium,and high doses of ISO were added to macrophages.Western Blot,RT-qPCR and flow cytometry were used to detect the expression of ISO on M2 markers and Yes-related protein(YAP)in M0 and M2 macrophages.In addition,a YAP inhibitor was added and its effect on macrophage polarization was examined.Results:In the M0 and M2 macrophages,the expression of M2 marker[mannose receptor(CD206)]and YAP in the cells increased after ISO-treated for 24 h(P<0.05,P<0.01).Flow cytometry also confirmed that the mean fluorescence intensity(MFI)of CD206 in macrophages was significantly increased after ISO treatment(P<0.05,P<0.01).In addition,YAP inhibitors can attenuate the effect of ISO on the up-expression of CD206 in macrophages(P<0.01).Conclusion:ISO acts as an immunomodulator and promotes the M2 polarization of macrophages,thereby playing an anti-inflammatory role via up-regulation of YAP.

关 键 词：异槲皮素 巨噬细胞极化 抗炎 Yes相关蛋白 免疫 

分 类 号：R285[医药卫生—中药学]