犀角地黄汤合方对DC激活ITP患者T细胞增殖分化的影响  

The effect of Xijiao Dihuang combined prescription on T cell differentiation induced by platelet antigen loaded DC in ITP patients

作　　者：杨武霞 刘宝山[2] 吴玉红[3] 李润杰 王梦晓 王爱迪[2] YANG Wuxia;LIU Baoshan;WU Yuhong;LI Runjie;WANG Mengxiao;WANG Aidi(Graduate School of Tianjin Medical University,Tianjin 300041,China;Department of Traditional Chinese Medicine,Tianjin Medical University General Hospital;Department of Hematology,Tianjin Medical University General Hospital)

机构地区：[1]天津医科大学研究生院,300041 [2]天津医科大学总医院中医科 [3]天津医科大学总医院血液科

出　　处：《天津医药》2022年第12期1233-1239,共7页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(82004334);天津市卫生健康委员会,天津市中医重点专科建设项目。

摘　　要：目的 观察犀角地黄汤合方含药血清对树突状细胞(DC)激活T细胞增殖分化的影响及其对免疫性血小板减少症(ITP)的作用机制。方法 将10只雄性SD大鼠随机分为含药血清组和空白血清组,每组5只,分别使用犀角地黄汤合方和蒸馏水连续灌胃3 d,腹主动脉取血制备含药血清和空白血清。分选15例健康志愿者及8例ITP患者CD4^(+)T细胞,将荷载血小板抗原的DC与CD4^(+)T细胞共培养,采用随机数字表法分为对照组,模型组,犀角地黄汤合方低、中、高剂量组。对照组为荷载血小板抗原的DC与健康志愿者CD4^(+)T细胞,模型组和犀角地黄汤合方低、中、高剂量组为荷载血小板抗原的DC与ITP患者的CD4^(+)T细胞,其中对照组和模型组加入大鼠空白血清,犀角地黄汤合方低、中、高剂量组分别加入体积分数为5%、10%及20%大鼠含药血清。流式细胞术(FCM)检测CD4^(+)T细胞的增殖情况,各组调节性T细胞(Treg)和效应性T细胞(Teff)的比例以及CD4^(+)T表面程序性死亡受体-1(PD-1)的表达量;酶联免疫吸附试验检测各组细胞上清液中促炎因子白细胞介素(IL)-2、干扰素-γ(IFN-γ)、IL-17以及抑炎因子转化因子-β(TGF-β)、IL-10的表达量。结果 与对照组比较,模型组CD4^(+)T细胞增殖百分比、Teff细胞比例以及促炎因子IL-2、IFN-γ和IL-17升高(P<0.05),Treg细胞比例、CD4^(+)T细胞表面PD-1表达量以及抑炎因子TGF-β、IL-10水平降低(P<0.05);与模型组比较,犀角地黄汤合方低、中、高剂量组CD4^(+)T细胞增殖百分比、Teff细胞比例依次降低,Treg细胞比例、CD4^(+)T细胞表面PD-1表达量和抑炎因子IL-10、TGF-β升高(P<0.05),促炎因子IL-2的量降低(P<0.05);与模型组相比,犀角地黄汤合方中、高剂量组IFN-γ和IL-17降低(P<0.05)。结论 犀角地黄汤合方尤其是高剂量组含药血清能够在体外调节ITP患者CD4^(+)T细胞的过度增殖分化及细胞因子的分泌,这可能是犀角地黄汤Objective To observe the effect of drug-containing serum of Xijiao Dihuang combined prescription(XJDHCP) on the proliferation and differentiation of dendritic cell(DC)-activated T cells and its mechanism on immune thrombocytopenia(ITP).Methods Ten male SD rats were randomly divided into the drug-containing serum group and the blank serum group,with 5 rats in each group.Rats were gavaged with XJDHCF and distilled water for 3 days.Bloodextraction from the abdominal aorta was prepared for drug-containing serum and blank serum.Fifteen healthy volunteers and8 ITP patients with CD4^(+)T cells were selected,and the DCs loaded with platelet antigens were co-cultured with CD4^(+)T cells and randomly divided into the normal control group,the model group and the low,medium and high(5%,10% and 20%)dose groups of XJDHCF.The normal control group consisted of platelet antigen-loaded DCs and CD4^(+)T cells from healthy volunteers,and the model group and the low,medium and high dose groups of XJDHCF were platelet antigen-loaded DCs and CD4^(+)T cells from ITP patients,in which the normal control group and the model group were added with rat blank serum,and the low,medium and high dose groups of XJDHCF were added with 5%,10% and 20% drug-containing serum.The proliferation of CD4^(+)T cells,the ratio of regulatory T cells(Treg),effector T cells(Teff) and the expression of programmed death receptor-1(PD-1) on the surface of CD4^(+)T were detected by flow cytometry(FCM).Enzyme-linked immunosorbent assay was used to detect the expression levels of pro-inflammatory factors IL-2,IFN-γ,IL-17 and anti-inflammatory factors TGF-β and IL-10 in the supernatant of each group.Results Compared with the control group,the percentage of CD4^(+)T cell proliferation,the proportion of Teff cells and amounts of pro-inflammatory factors IL-2,IFN-γ and IL-17 were significantly higher in the model group(P <0.05),and the proportion of Treg cells,the amount of PD-1 expression on the surface of CD4^(+)T cells and amounts of anti-inflammatory factors TGF-�

关 键 词：血小板减少 树突状细胞 CD4阳性T淋巴细胞 疾病模型 动物 犀角地黄汤合方 

分 类 号：R558.2[医药卫生—血液循环系统疾病]