芪黄益气摄血方对免疫性血小板减少症模型小鼠的治疗作用及抗炎机制研究

Therapeutic effect and anti-inflammatory mechanism of Qihuang Yiqi Shexue Formula in ITP model mice

目的探讨芪黄益气摄血方对免疫性血小板减少症(ITP)模型小鼠的治疗作用及抗炎机制。方法50只BALB/c小鼠随机分为正常组,模型组,芪黄益气摄血方高、低剂量组和强的松组,每组10只。采用豚鼠抗小鼠血小板血清(APS)腹腔注射方法,建立ITP模型;注射APS后的第8天开始给药,连续灌胃14 d。观察小鼠一般情况;检测外周血血小板计数(PLT)和血红蛋白(Hb)浓度变化;Elisa法检测血清血小板生成素(TPO)、促炎因子白介素-2(IL-2)、IL-6、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)及抑炎因子IL-4、IL-10、转化生长因子-β1(TGF-β1)水平;分离脾脏、胸腺,称重,计算脏器指数;取胸骨做骨髓涂片,显微镜下进行骨髓巨核细胞分类。结果与正常组比较,模型组小鼠PLT、Hb、TPO、IL-4、IL-10、TGF-β1显著降低(P<0.05或P<0.01),骨髓产板巨核细胞数明显减少(P<0.01),血清IL-2、IL-6、TNF-α、IFN-γ以及脾脏和胸腺指数显著升高(P<0.05或P<0.01);与模型组比较,各给药组PLT、TPO、IL-4、IL-10、TGF-β1水平明显升高(P<0.05或P<0.01),骨髓产板巨核细胞数明显增加(P<0.01),血清IL-2、IL-6、TNF-α、IFN-γ以及脾脏和胸腺指数显著降低(P<0.05或P<0.01),仅高剂量组Hb显著增加(P<0.05);与低剂量组比较,高剂量和强的松组PLT、IL-4、TGF-β1显著升高(P<0.05或P<0.01),TNF-α显著降低(P<0.05或P<0.01),仅强的松组IL-6显著降低(P<0.05),高剂量组IL-10显著升高(P<0.05),其余指标无统计学差异(P>0.05)。结论芪黄益气摄血方可通过调节ITP模型小鼠促炎因子和抑炎因子的分泌失衡,增加骨髓产板巨核细胞和TPO水平,发挥治疗ITP的作用。

Objective To explore the therapeutic effect and anti-inflammatory mechanism of Qihuang Yiqi Shexue Formula in immune thrombocytopenia(ITP)model mice.Methods Fifty BALB/c mice were randomly divided into the normal group,model group,high and low dose groups of Qihuang Yiqi Shexue Formula,and the prednisone group,with 10 mice in each.The intraperitoneal injection of guinea pig anti-mouse platelet serum(APS)was used to establish an ITP mouse model.On the 8th day after the injection of APS,each group was given intragastric administration for 14 consecutive days.The general condition of each group of mice was observed,changes in peripheral blood platelet count(PLT)and hemoglobin(Hb)concentrations were detected,and the Elisa method was used to detect serum levels of thrombopoietin(TPO),pro-inflammatory factor interleukin-2(IL-2),IL-6,tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),anti-inflammatory factor IL-4,IL-10,and transforming growth factor-β1(TGF-β1).The spleen and thymus tissues were separated,weighed,and the organ index was calculated.The sternum was taken as a bone marrow smear,and the bone marrow megakaryocytes were classified under a microscope.Results Compared with the normal group,the levels of PLT,Hb,TPO,IL-4,IL-10 and TGF-β1 in the model group significantly decreased(P<0.05 or P<0.01),the number of megakaryocytes in the bone marrow plate became significantly smaller(P<0.01),and serum levels of IL-2,IL-6,TNF-αand IFN-γwere significantly decreased(P<0.05 or P<0.01).Compared with the model group,the levels of PLT,TPO,IL-4,IL-10,and TGF-β1in each administration group were significantly increased(P<0.05 or P<0.01),so was the number of megakaryocytes in the bone marrow plate(P<0.01),but serum levels of IL-2,IL-6,TNF-αand IFN-γwere significantly decreased(P<0.05 or P<0.01).Compared with the low-dose group,the levels of PLT,IL-4 and TGF-β1 were significantly increased in the high dose group and the prednisone group,but TNF-αwas significantly decreased(P<0.05 or P<0.01).There was no statistically significant difference in other indicators(P>0.05).Conclusion Qihuang Yiqi Shexue Formula is effective for ITP by regulating the unbalanced secretion of pro-inflammatory factors and anti-inflammatory factors in ITP model mice,increasing the levels of bone marrow-producing megakaryocytes and TPO.