白芍总苷对肥大细胞脱颗粒的影响  

Effects of Total Glucosides of Paeony on Degranulation of Mast Cells

作　　者：陈丽明 黄彬 刘用诚 林久茂 林明和 CHEN Liming;HUANG Bin;LIU Yongcheng;LIN Jiumao;LIN Minghe(The Second People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350003,China;Institute of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学附属第二人民医院,福建福州350003 [2]福建中医药大学中西医结合研究院,福建福州350122

出　　处：《福建中医药》2024年第7期11-16,共6页Fujian Journal of Traditional Chinese Medicine

基　　金：福建中医药大学2022年校管临床专项课题(XB2022030)。

摘　　要：目的研究白芍总苷(TGP)对肥大细胞(MCs)脱颗粒的影响。方法将对数生长期的小鼠肥大细胞瘤细胞P815(下称P815细胞)分成12组,分别采用0、25、50、100、200、400μg/mL TGP溶液干预24 h和48 h,以筛选药物的安全干预浓度和时间。取对数生长期P815细胞分为对照组、模型组和低、中、高剂量组,低、中、高剂量组分别加入2 mL浓度为25、50、100μg/mL TGP溶液干预24 h,对照组及模型组加入与上述TGP溶液等体积的完全培养基孵育,随后除对照组外,其他各组加入浓度为20μg/mL的致敏剂C48/80刺激1 h诱导P815细胞脱颗粒。采用甲苯胺蓝染色观察5组细胞形态变化;ELISA法检测5组细胞中β-氨基己糖苷酶和组胺水平,荧光探针法检测细胞内钙离子相对荧光强度,转录组学测序分析模型组与对照组、模型组与中剂量组的差异表达基因,并对差异表达基因进行KEGG通路富集分析。结果与对照组比较,模型组细胞体积变大,形态不完整,边缘模糊,周围散在颗粒,细胞质淡染,细胞中β-氨基己糖苷酶和组胺水平显著增加,细胞内钙离子荧光强度明显增强(P<0.05);与模型组比较,中、高剂量组均可改善细胞脱颗粒形态变化,降低组胺和β-氨基己糖苷酶水平以及细胞内钙离子相对荧光强度(P<0.05),而低剂量组上述指标变化不明显(P>0.05),中剂量组与高剂量组比较,β-氨基己糖苷酶和组胺水平以及钙离子的荧光强度差异无统计学意义(P>0.05)。模型组与对照组以及模型组与中剂量组的差异表达基因主要富集在MAPK、JAK-STAT、IL-17、TNF、NF-κB信号通路。结论TGP可有效抑制C48/80诱导的P815细胞脱颗粒,改善细胞形态变化,抑制β-氨基己糖苷酶、组胺以及钙离子的释放,MAPK、JAK-STAT、IL-17、TNF、NF-κB信号通路可能是TGP抗过敏反应的关键靶通路。Objective:To investigate the impact of Total Glucosides of Paeony(TGP)on the degranulation of mast cells(MCs).Methods:P815 mastocytoma cells were divided into 12 groups and treated with TGP solution at concentrations of 0,25,50,100,200,and 400μg/mL for 24 h and 48 h to screen for safe intervention conditions.P815 cells in the logarithmic growth phase were divided into control group,model group,and TGP low-,medium-,and high-dose groups.The low-,medium-,and high-dose groups were incubated with 2 mL TGP solutions at 25,50,and 100μg/mL for 24 h,respectively,while the control and model groups were incubated with complete culture-medium of the same volume as the above TGP solutions.Subsequently,except for the control group,other groups of P815 cells were stimulated with 20μg/mL of the sensitizer compound 48/80 for 1 h to induce degranulation.Toluidine blue staining was used to observe cell morphological changes;ELISA was used to detect the levels ofβ-hexosaminidase and histamine;and a fluorescent probe method was used to detect the fluorescence intensity of calcium ions.Transcriptomics was used to screen for differentially expressed genes between the model group and the control group,as well as between the model group and the medium-dose group.Besides that,those genes were also used for KEGG pathway analysis.Results:Compared with the control group,the cell volumes of the model group increased with incomplete morphology,and blurred edges,scattered granules,pale cytoplasm,and levels of histamine andβ-hexosaminidase significantly increased,as well as a enhancement in intracellular calcium ion fluorescence intensity(P<0.05).Compared with the model group,both the medium-and high-dose groups could improve cellular morphology of degranulation,reduce levels of histamine andβ-hexosaminidase,and decrease fluorescence intensity of calcium ions(P<0.05),while the low-dose group showed no significant changes(P>0.05).There was no significant difference inβ-hexosaminidase,histamine,and calcium ion fluorescence intensity between

关 键 词：白芍总苷 肥大细胞 脱颗粒 过敏 转录组学 

分 类 号：R285[医药卫生—中药学]