去甲斑蝥素联合环磷酰胺对H22荷瘤小鼠免疫功能的影响

Effects of norcantharidin on the immunological function in tumor-bearing mice

目的：探讨去甲斑蝥素联合环磷酰胺对荷瘤小鼠免疫功能的影响。方法：采用小鼠体内培养H22腹水肝癌细胞，接种健康小鼠，建立荷瘤小鼠模型，并给予去甲斑蝥素联合环磷酰胺治疗，观察计算脾脏指数、胸腺指数，CCK-8检测荷瘤小鼠经去甲斑蝥素联合环磷酰胺治疗后的脾脏淋巴细胞转化实验，流式检测经去甲斑蝥素联合环磷酰胺治疗后荷瘤小鼠T细胞亚群CD3＋、CD4＋、CD8＋百分率，骨髓有核细胞凋亡率及细胞周期。结果：与环磷酰胺组比较，去甲斑蝥索联合环磷酰胺处理组的脾脏指数有升高趋势；在淋巴细胞转化实验中，不同剂量的去甲斑蝥素联合环磷酰胺均可提高淋巴细胞转化率，且与去甲斑蝥素的剂量呈正相关。低剂量的去甲斑蝥素联合环磷酰胺可有效提高T细胞亚群的百分比，CD3＋（％）为11．4±3．13，CD4＋（%）为7．3±2．94，CD8＋（％）为2．88±0．51，与环磷酰胺组比较P〈0．05，与去甲斑蝥素的浓度呈负相关。随着去甲斑蝥素剂量的增加，骨髓有核细胞凋亡率也逐渐升高。结论：低剂量去甲斑蝥素（1mg/kg）联合环磷酰胺可以降低单纯环磷酰胺化疗药对免疫系统的抑制作用，为临床用药剂量提供参考依据。

OBJECTIVE： To explore the effects of noreantharidin on the immunological function in tumor-bearing mice. METHODS：The models of tumor-bearing mice were established by inoculating the healthy mice with murine hepatocareinoma cell line H22 cultrued in vivo, and then treated by norcantharidin. The ratio of spleen index and thymus index were calculated. The proliferation activity of T lymphoeytes in the mice after 14 days being treated with norcantharidiin was detected by CCK-8.The percentage of CD3＋, CD4＋,CD8＋ were measured by flow Cytometry. Apoptosis and cell cycle were detected by AnnexinV/propidium iodide staining.. RESULTS：Compared with the eyclophosphamide group, noreantharidin and eyclophosphamide can increase the spleen index of the mice. On lymphocyte transformation, norcantharidin and eyclophosphamide both can enhance the ratio. Low-dose noreantharidin and eyelophospbamide can enhance the ratio of T lymphocyte subsets significantly, the percentage of CD3＋ was up tol 1.4 ± 3.13, CD4＋ was 7.3 ± 2.94,and CD8＋ was up to 2.88 ± 0.51 （compared with the eyclophosphamide group, P〈0.05）. The apoptosis percentage of bone marrow ceils gradually increased depending on the concentrationg of norcantharidin.CONCLUTION：Low-dose noreantharidin （1mg/kg） can enhance the immunological function of tumor-bearing mice.