藤黄酚对小鼠脾淋巴细胞增殖及细胞因子分泌的抑制作用  

Inhibitory Effect of Garcinol on Proliferation and Secretion of Cytokines of Splenic Lymphocytes in Mice

作　　者：王明智 徐静[2] 陈元杰 岑举人[1] WANG Mingzhi;XU Jing;CHEN Yuanjie;CEN Juren(School of Life and Health,Hainan University,Haikou,570228;Hainan Provincial Fine Chemical Engineering Research Center,College of Chemical Engineering and Technology,Hainan University,Haikou,570228)

机构地区：[1]海南大学生命健康学院,海口570228 [2]海南大学化学工程与技术学院,海南省精细化工工程技术研究中心,海口570228

出　　处：《基因组学与应用生物学》2025年第2期192-200,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(82160675);2022年度海南大学协同创新中心项目(XTCX2022STB01)共同资助。

摘　　要：本研究旨在探讨藤黄酚(garcinol,Gar)对小鼠(Mus musculus)脾淋巴细胞的免疫抑制作用及其潜在机制。本研究采用不同剂量(2、4、6、8、10μmol/L)的Gar作用于刀豆蛋白A(concanavalin A,Con A)诱导的小鼠脾淋巴细胞,采用CCK-8法评估脾淋巴细胞活力,采用流式细胞术测定脾淋巴细胞中CD4^(+)、CD8^(+)T细胞的比例,采用ELISA法检测细胞上清液中细胞因子IL-2、IFN-γ及IL-4的含量,采用钙调蛋白磷酸酶(calcineurin,CaN)活性检测试剂盒检测细胞内CaN的活力。此外,运用免疫荧光染色法观察细胞NFAT蛋白核易位情况,并通过Western blot检测细胞NFAT、p65、IκBα、p-p65、p-IκBα的蛋白水平。结果显示,Gar在非细胞毒性的浓度下能显著抑制Con A诱导的脾淋巴细胞增殖,其半抑制浓度(half inhibitory concentration,IC50)为7.05μmol/L。此外,Gar可剂量依赖性降低Con A诱导的脾淋巴细胞中CD4^(+)T细胞的比例和IL-2、IFN-γ、IL-4的分泌水平,降低CaN酶活力和NFAT、p-p65、p-IκBα的蛋白水平,并抑制NFAT核易位。本研究发现Gar能抑制Con A诱导的小鼠脾淋巴细胞增殖及细胞因子分泌,其作用机制可能与抑制CaN/NFAT和NF-κB信号通路有关。This study aims to investigate the immunosuppressive effects of garcinol(Gar)on mice(Mus musculus)splenic lymphocytes and its underlying mechanism.In this study,splenic lymphocytes induced by concanavalin A(Con A)were treated with various concentrations of Gar(2,4,6,8,10μmol/L).The CCK-8 assay was used to assess the viability of splenic lymphocytes,and flow cytometry was employed to determine the proportions of CD4^(+)and CD8^(+)T cells within the splenic lymphocytes.The contents of cytokines IL-2,IFN-γ,and IL-4 in the cell culture supernatant were detected by ELISA,and the activity of calcineurin(CaN)was detected by a CaN activity assay kit.In addition,the nuclear translocation of the NFAT protein was observed by immunofluorescence staining,and the protein levels of NFAT,p65,IκBα,p-p65,and p-IκBαwere detected by Western blot analysis.The results showed that Gar significantly inhibited Con A-induced splenic lymphocyte proliferation at non-cytotoxic concentrations,with a half inhibitory concentration(IC_(50))of 7.05μmol/L.Moreover,Gar dose-dependently reduced the proportion of CD4^(+)T cells and the secretion levels of IL-2,IFN-γand IL-4.It also inhibited NFAT nuclear translocation and decreased the protein levels of NFAT,p-p65,and p-IκBα,as well as the enzymatic activity of CaN in Con A-induced splenic lymphocytes.The present study found that Gar can inhibit Con A-induced splenic lymphocyte proliferation and cytokine secretion in mice,and its mechanism may be related to the inhibition of the CaN/NFAT and NF-κB signaling pathways.

关 键 词：藤黄酚 T淋巴细胞增殖 免疫抑制剂 CaN/NFAT NF-ΚB 

分 类 号：R285[医药卫生—中药学]