小檗胺和小檗碱影响嗜酸性粒细胞凋亡及活性的体外研究  

Effects of berbamine and berberine on the apoptosis and activity of eosinophils in vitro

作　　者：程序 谷春 安成[1] 侯雪筠[1] 费佳欣 刘贵建[1] 李勇 庞博[1] CHENG Xu;GU Chun;AN Cheng;HOU Xuejun;FEI Jiaxin;LIU Guijian;LI Yong;PANG Bo(Clinical Laboratory,Guang’anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China;Institute of Pharmacy,Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国中医科学院广安门医院检验科,北京100053 [2]北京协和医学院药物研究所,100050

出　　处：《免疫学杂志》2024年第5期411-417,424,共8页Immunological Journal

摘　　要：目的探索小檗碱(berberine,BBR)和小檗胺(berbamine,BA)对嗜酸性粒细胞凋亡及活性的作用,为嗜酸性粒细胞相关疾病的治疗提供新的思路。方法取入院患者的抗凝全血,经过磁珠分选技术进行嗜酸性粒细胞纯化,流式细胞术检测比较细胞纯化率。培养纯化的嗜酸粒细胞并给予不同浓度的BBR、BA干预。使用免疫荧光显微镜观察嗜酸性粒细胞凋亡情况;流式细胞术检测嗜酸性粒细胞凋亡率、活性氧释放情况、线粒体膜电位情况及CD11b、CD62L、CD63分子表达情况。结果10μmol/L BBR对嗜酸性粒细胞在18 h、24 h、48 h显示有显著的抗凋亡作用;而100μmol/L和150μmol/L BBR对嗜酸性粒细胞在24 h、48 h显示有显著的促凋亡作用,尤其是早期凋亡。5μmol/L、15μmol/L和25μmol/L BA对嗜酸性粒细胞在24 h、48 h显示有显著的促凋亡作用,尤其是晚期凋亡。在嗜酸性粒细胞中,BA浓度依赖性干预活性氧的产生及线粒体膜电位的下降,在25μmol/L达到最大效果,而BBR的干预没有使线粒体膜电位、活性氧释放情况发生变化。25μmol/L BA预处理对人嗜酸性细胞趋化因子-2干预后嗜酸粒细胞CD11b表达升高和CD62L表达降低有显著的抑制作用。结论BBR对嗜酸性粒细胞的数量具有双向调节作用,BA具有促嗜酸性粒细胞凋亡、抑制嗜酸性粒细胞激活的作用,二者在调节嗜酸性粒细胞相关疾病进展方面具有一定应用价值,其调节机制值得进一步探讨。This study was performed to explore the effects of berberine(BBR)and berbamine(BA)on the apoptosis and activity of eosinophils,and to provide new ideas for the treatment of eosinophil-related diseases.Anticoagulated whole blood was taken from hospitalized patients,and eosinophils were purified by magnetic bead sorting technology,and the cell purification rate was compared by using flow cytometry.The purified eosinophils were cultured and stimulated with different concentrations of BBR and BA,and the apoptosis of eosinophils was observed using immunofluorescence microscopy.Flow cytometry was used to detect the apoptosis rate,reactive oxygen species(ROS)release,mitochondrial membrane potential,and CD11b,CD62L and CD63 molecular expression of eosinophils.Data showed that 10μmol/L BBR displayed significant anti-apoptosis effect on eosinophils at 18 h,24 h,48 h,however,100μmol/L and 150μmol/L BBR displayed significant apoptosis-promoting effects on eosinophils at 24 h,48 h,especially in the early stage.BA at 5μmol/L,15μmol/L and 25μmol/L showed significant apoptosis-promoting effects on eosinophils at 24 h,48 h,especially in the late stage.In eosinophils,BA stimulated the production of ROS and the decrease of mitochondrial membrane potential in a concentrationdependent manner,reaching the maximum effect at 25μmol/L,while BBR stimulation did not change mitochondria and ROS.Pretreatment with 25μmol/L BA significantly inhibited the increase of CD11b expression and the decrease of CD62L expression in eosinophils after eotaxin-2 stimulation.In conclusion,berberine has a bidirectional regulatory effect on the quantity of eosinophils,while berbamine promotes eosinophil apoptosis and inhibits eosinophil activation.Both compounds hold significant potential for regulating the progression of eosinophil-associated diseases,and their regulatory mechanisms need further investigation.

关 键 词：嗜酸性粒细胞 小檗碱 小檗胺 凋亡 

分 类 号：R743.34[医药卫生—神经病学与精神病学]