益髓颗粒对ITP小鼠脾脏CD_4^+CD_(25)^+Foxp_3^+Treg细胞及其相关因子影响的研究

目的建立免疫性血小板减少性紫癜(ITP)小鼠动物模型,观察益髓颗粒对模型小鼠脾脏CD4+CD25+Foxp3+Treg细胞(Treg)及血清TGF-β1的影响,探讨益髓颗粒的免疫调节机制。方法 40只Balb/c小鼠随机分为正常组、模型组、泼尼松治疗组、益髓颗粒治疗组,除正常组外,其余各组均隔日一次按100μl/20g腹腔注射1:4豚鼠抗小鼠血小板血清(GP-APS)建立ITP小鼠模型,造模第8天开始各组均按0.2ml/10g·d-1体积灌胃,正常组、模型组灌服生理盐水,益髓颗粒组治疗按0.3g/10g·d-1灌胃,连续8天后眼球取血分离血清,ELISA法检测TGF-β1,取脾制备脾单细胞悬液,用流式细胞仪检测脾Treg。结果模型组小鼠Treg细胞比例、TGF-β1水平明显低于正常组;泼尼松及益髓颗粒治疗组Treg细胞比例及TGF-β1水平均高于模型组,但Treg仍未达到正常水平,与正常组比仍存在差异,而TGF-β1与正常组比差异不明显无统计学意义。结论 ITP小鼠Treg细胞比例减少、TGF-β1水平降低可能与ITP的细胞免疫失调相关,益髓颗粒可通过提高ITP小鼠Treg细胞的比例及血清TGF-β1水平而对ITP的细胞免疫失调起调节作用。

Objective To observe the influence of Yisui granules on CD4 ＋ CD 25 ＋ Foxp 3 ＋ regulatory T cells（Treg）in spleen and the level of TGF-β1 in serum in immune thrombocytopenic purpura（ITP）mouse model and discuss Yisui granules＇s immune regulatory mechanisms.Methods 40 Balb/ c mice were randomly divided into normal group,model group,Prednisolone acetate group and Yisui granules group.ITP mouse model were made by injected intraperitoneally 1：4 guinea pig-antimouse platelet serum（GP-APS）at a dose of 100μl/20g every other day except the normal group.The 8th day establishing the model,each group were inrtagastrically at a dose of 0.2ml/10g·d-1 volume.Normal group and model group fed with normal saline,Prednisone acetate and YiSui granules were inrtagastrically administered eight days at a dose of 0.0113mg/10g·d-1,0.3g/10g·d-1 respectively. The levels of TGF-β1 in serum were detected by ELISA and the Treg cells were detected by flow cytometry（FCM）in spleen single cell suspension. Results The proportion of Treg cells and the levels of TGF-β1 was significantly lower in model group as compared to normal group.The proportion of Treg cells and the levels of TGF-β1 was higher in Prednisone group and YiSui granules group as compared to model group.However,the proportion of Treg cells was lower in Prednisone group and YiSui granules group than that in normal group,there is no significant difference of the levels of TGF-β1 in Prednisone group and YiSui granules group as compared to normal group.Conclusion The decrease of Treg cells and the levels of TGF-β1 in ITP mouse model maybe related to cellular immunity disorder.Yisui granules can regulate the cellular immunity disorder of ITP by increasing proportion of Treg cells and the level TGF-β1 in ITP mouse model.