黄芩素对Naive CD4^(+)T细胞向Th1和Treg细胞分化的影响  

Effect of baicalein on the differentiation of Naive CD4^(+)T cells into Th1 cells and Treg cells

作　　者：佟慧敏 王爽[1] 李玲[1] TONG Huimin;WANG Shuang;LI Ling(School of Basic Medical,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院,山东青岛266071

出　　处：《精准医学杂志》2025年第1期8-12,18,共6页Journal of Precision Medicine

基　　金：山东省自然科学基金资助项目(ZR2020MH416)。

摘　　要：目的探讨黄芩素对Naive CD4^(+)T细胞向Th1和Treg细胞分化的影响。方法从6~8周龄的BALB/c雄性小鼠脾脏中分选出Naive CD4^(+)T细胞,并将其分为A~F组。A组细胞接种在包被有抗小鼠CD3抗体的孔板中,在含抗小鼠CD28抗体的培养基中培养72 h;B组细胞在A组基础上再加入白细胞介素-2(IL-2)、IL-12和抗小鼠IL-4抗体;C组细胞在B组的基础上再加入黄芩素。D组细胞的培养体系与A组相同;E组细胞在D组的基础上再加入IL-2和转化生长因子-β(TGF-β);F组细胞在E组的基础上再加入黄芩素。通过流式细胞术检测A~C组中Th1细胞的比例和D~F组中Treg细胞的比例;通过RT-qPCR检测A~C组细胞中T盒子转录因子(T-bet)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)的相对表达水平和D~F组中叉头框蛋白P3(Foxp3)和IL-10的相对表达水平;通过免疫荧光染色检测A~C组中Th1细胞阳性率和D~F组中Treg细胞阳性率。结果流式细胞术检测结果显示,C组Th1细胞比例显著低于B组(F=46.91,q=7.95,P<0.05),F组Treg细胞比例显著高于E组(F=145.20,q=8.64,P<0.05)。RT-qPCR检测结果显示,C组细胞中T-bet、IFN-γ、TNF-α的相对表达水平显著低于B组(F=15.77~862.50,q=7.05~26.34,P<0.05),F组细胞中Foxp3和IL-10的相对表达水平显著高于E组(F=38.25、49.24,q=4.65、7.64,P<0.05)。免疫荧光结果显示,C组Th1细胞阳性率显著低于B组(F=11.64,q=4.50,P<0.05),F组Treg细胞阳性率显著高于E组(F=20.31,q=4.64,P<0.05)。结论黄芩素对Naive CD4^(+)T细胞向Th1细胞分化具有抑制作用,对Naive CD4^(+)T细胞向Treg细胞分化具有促进作用。Objective To investigate the effect of baicalein on the differentiation of Naive CD4^(+)T cells into Th1 cells and Treg cells.Methods Naive CD4^(+)T cells were isolated from the spleen of male BALB/c mice aged 6-8 weeks and were divided into groups A to F.The cells in group A were inoculated into a well plate coated with anti-mouse CD3 antibody and were cultured in a medium containing anti-mouse CD28 antibody for 72 h;in addition to the treatment in group A,the cells in group B were treated with interleukin-2(IL-2),IL-12,and anti-mouse IL-4 antibodies;in addition to the treatment in group B,the cells in group C were further treated with baicalein.The cells in group D were cultured in the same culture system as group A;in addition to the treatment in group D,the cells in group E were further treated with IL-2 and transforming growth factor-β(TGF-β);in addition to the treatment in group E,the cells in group F were further treated with baicalein.Flow cytometry was used to measure the percen-tage of Th1 cells in groups A to C and the percentage of Treg cells in groups D to F;RT-qPCR was used to measure the relative expression levels of T-box transcription factor(T-bet),interferon-γ(IFN-γ),and tumor necrosis factor-α(TNF-α)in groups A to C and the relative expression levels of forkhead box P3(Foxp3)and IL-10 in groups D to F;immunofluorescent staining was used to measure the positive rate of Th1 cells in groups A to C and the positive rate of Treg cells in groups D to F.Results Flow cytometry showed that group C had a significantly lower percentage of Th1 cells than group B(F=46.91,q=7.95,P<0.05),and group F had a significantly higher percentage of Treg cells than group E(F=145.20,q=8.64,P<0.05).RT-qPCR showed that group C had significantly lower relative expression levels of T-bet,IFN-γ,and TNF-αthan group B(F=15.77-862.50,q=7.05-26.34,P<0.05),and group F had significantly higher relative expression levels of Foxp3 and IL-10 than group E(F=38.25,49.24,q=4.65,7.64,P<0.05).Immunofluorescence assay showed that g

关 键 词：黄芩苷 TH1细胞 T淋巴细胞 调节性 细胞分化 体外研究 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]