健脾益气摄血方对模型小鼠血小板减少性紫癜的改善作用  

Improvement Effect of Jianpi Yiqi Shexue Formula on Thrombocytopenic Purpura in Model Mice

作　　者：万敏[1] 谭晶晶[1] 周林[1] WAN Min;TAN Jingjing;ZHOU Lin(Department of Clinical Laboratory,Wuhan Hospital of Traditional Chinese Medicine,Wuhan,Hubei,China 430014)

机构地区：[1]湖北省武汉市中医医院检验科,湖北武汉430014

出　　处：《中国药业》2022年第6期55-60,共6页China Pharmaceuticals

基　　金：湖北省武汉市医学科研项目[WZ19C37]。

摘　　要：目的探讨健脾益气摄血方对模型小鼠血小板减少性紫癜(ITP)的改善作用。方法将60只BALB/c小鼠随机分为空白对照组(等体积生理盐水)、模型组(等体积生理盐水)、醋酸泼尼松组(5 mg/kg)及健脾益气摄血方低、中、高剂量组(6.96,13.91,27.82 g/kg),各10只。除空白对照组外,其余各组均采用豚鼠抗小鼠血小板血清腹腔注射,以复制小鼠ITP模型。建模成功后,各组小鼠均灌胃给予相应药物或生理盐水,每日1次,连续14 d。各组小鼠给药后7,14 d时取眼眶静脉血,检测外周血血小板计数;苏木素-伊红(HE)染色,光镜下观察小鼠脾脏及肾脏组织病理形态学;采用流式细胞仪检测小鼠脾脏中辅助性T细胞17(Th17)/调节性T细胞(Treg)比例;小鼠眼球取血,采用酶联免疫吸附(ELISA)法检测小鼠血清中白细胞介素17(IL-17)、白细胞介素6(IL-6)、转化生长因子-β_(1)(TGF-β_(1))及白细胞介素10(IL-10)水平;采用实时定量聚合酶链反应法及Westernblot法分别检测脾脏组织中叉头样转录因子3(Foxp3)及孤独核受体(RORγt)mRNA和蛋白的表达水平。结果与模型组比较,各用药组小鼠外周血血小板计数均明显增多;光镜下脾脏中异常骨髓巨核细胞数量不同程度减少,且脾脏Th17细胞明显减少,Treg细胞明显增多;血清IL-10水平明显降低,IL-17,IL-6,TGF-β_(1)水平均明显升高;脾脏组织中RORγt mRNA及蛋白表达水平均显著降低,Foxp3 mRNA及蛋白表达水平均显著升高(P<0.05)。结论健脾益气摄血方对模型小鼠ITP有明显改善作用,其作用机制可能与调节Th17/Treg细胞免疫失衡有关。Objective To investigate the improvement effect of Jianpi Yiqi Shexue Formula on immune thrombocytopenic purpura(ITP)in model mice.Methods 60 BALB/c mice were randomly divided into blank control group(equal volume of normal saline),model group(equal volume of normal saline),prednisone acetate group(5 mg/kg),and Jianpi Yiqi Shexue Formula low-,medium-and high-dose groups(6.96,13.91,27.82 g/kg),with 10 mice in each group.Except for those in the blank control group,the ITP model mice were established by intraperitoneal injection of guinea pig anti-mouse platelet serum(GP-APS).After successful modeling,mice in each group were given corresponding drugs or normal saline by gavaging once a day for 14 d.Blood was taken from orbital vein of mice at 7 d and 14 d after administration,and peripheral blood platelet count was measured.Hematoxylin-eosin(HE)staining was used to observe the histopathophology of spleen and kidney of mice under light microscope.The ratio of helper T cell 17(Th17)/regulatory T(Treg)cell in the spleen of rats was detected by the flow cytometry.Blood samples were taken from the eyeballs of mice,and the levels of interleukin-17(IL-17),interleukin-6(IL-6),transforming growth factorβ_(1)(TGF-β_(1))and interleukin-10(IL-10)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of forkhead like transcription factor 3(Foxp3)and retinoic receptor-related orphan receptor(RORγt)mRNA and protein in spleen were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot,respectively.Results Compared with those in the model group,the number of platelets in peripheral blood of mice in each treatment group increased significantly,the number of abnormal bone marrow megakaryocytes in spleen decreased in different degrees under light microscope,the number of Th17 cells in spleen decreased significantly,the number of Treg cells increased significantly,the IL-10 level of serum decreased significantly,the levels of IL-17,IL-6 and TGF-β_(1)increased significa

关 键 词：健脾益气摄血方 血小板减少性紫癜 小鼠 辅助性T细胞17 调节性T细胞 细胞免疫失衡 

分 类 号：R932[医药卫生—生药学] R285.5[医药卫生—药学]