益髓颗粒影响ITP小鼠脾淋巴细胞凋亡及Bcl-2、Bax蛋白表达的研究

目的]观察益髓颗粒对免疫性血小板减少性紫癜(ITP)小鼠脾淋巴细胞凋亡及Bcl-2、Bax蛋白表达的影响。方法]40只Balb/c小鼠随机分为正常组、模型组、泼尼松治疗组和益髓颗粒治疗组,除正常组外,其余各组均隔日1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,于造模第8天开始,各组均按0.2mL/10g·d~(-1)体积灌胃,其中,正常组、模型组灌服生理盐水,泼尼松按0.0113mg/10g·d~(-1)灌胃,益髓颗粒按0.3g/10g·d~(-1)灌胃,连续8天后处死动物,无菌取脾,石蜡包埋,切片4μm,用原位末端标记(TUNEL)法检测脾淋巴细胞凋亡,免疫组化法检测Bcl-2、Bax蛋白表达。结果]与正常组比较,模型组脾淋巴细胞凋亡减少,Bcl-2蛋白表达增多,Bax蛋白表达减少(P<0.05);与模型组比较,泼尼松及益髓颗粒组脾淋巴细胞凋亡增多,Bcl-2蛋白表达减少,Bax蛋白表达增多(P<0.05)。结论]ITP小鼠存在免疫功能失调可能与淋巴细胞凋亡及Bcl-2、Bax蛋白表达异常相关。益髓颗粒可通过调节淋巴细胞凋亡及Bcl-2、Bax蛋白表达而对ITP免疫失调起调节作用。

Study the Effects of YiSui Granules on Lymphocyte Apoptosis and Bcl2、Bax Protein Expression of Spleen in ITP Mouse Model

Objective] To observe the effect of Yisui granules on lymphocyte apoptosis and of Bcl- 2,Bax protein expression of spleen in immune thrombocytopenic purpura mouse model. 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^-l respectively. The spleen were embedded in paraffin and sliced 4μm, lymphocyte apoptosis were detected by TUNEL,Bcl-2 and Bax protein expression were detected by immunohistochemical. Results ] Compared to normal group, spleen lymphocyte apoptosis decreased, Bcl -2 protein expression increased, Bax protein expression decreased in model group （ P 〈 0.05 ）. Compared to model group, spleen lymphocyte apoptosis incrcased,Bcl-2 protein expression decreased, Bax protein expression increased in Prednisone group and YiSui granules group （ P 〈 0.05 ）. Conclusion ] There is immune dysfunction in ITP mouse, and the lymphocyte apoptosis and BCL -2, BAX protein expression abnormal may involve in ITP patho- genesis. YiSui granules can regulate the immune disorders of ITP by regulating lymphocyte apoptosis and Bcl - 2, Bax protein expression.