复方黄芪口服液联合化疗药物抗肿瘤效应机制研究

The study on mechanism of the anti-tumor effect of Fufang Huangqi Oral Solution combined chemotherapy

关键词:  复方黄芪口服液  H22荷瘤小鼠  Ⅰ型辅助性T细胞  调节性T细胞

Key words:  Fufang Huangqi Oral Solution  H22 tumor-bearing mice  Th1 cell  Treg cell

摘要:

[目的] 研究复方黄芪口服液联合化疗药物抗肿瘤效应及作用机制，为复方黄芪口服液抗肿瘤免疫疗法和临床应用提供依据。[方法] 构建H22荷瘤小鼠模型，随机分为荷瘤模型组、复方黄芪口服液组、环磷酰胺组和联合用药组。给药16 d后，分离肿瘤组织，评价抗肿瘤效应；血细胞分析仪测定外周血白细胞、中性粒细胞和单核细胞数量；四甲基偶氮唑蓝（MTT）法检测荷瘤小鼠淋巴细胞增殖功能；流式细胞术检测脾淋巴细胞Ⅰ型辅助性T细胞（CD4⁺IFN-γ⁺T细胞）和调节性T细胞（CD4⁺Foxp3⁺Treg细胞）表达水平。[结果] 与单用环磷酰胺相比，复方黄芪口服液联合环磷酰胺对肿瘤抑制率无明显影响，能够显著提高荷瘤小鼠脾淋巴细胞的增殖能力，差异有统计学意义（P<0.01）；复方黄芪口服液联合环磷酰胺进行干预后，可增加白细胞和中性粒细胞数量（P<0.01或P<0.05），上调CD4⁺IFN-γ⁺T细胞比例，下调具有免疫抑制作用的CD4⁺Foxp3⁺Treg细胞表达。[结论] 复方黄芪口服液联合化疗药物具有显著抗肿瘤免疫效应，其机制可能与促进Th1细胞反应和下调Treg细胞表达，从而提高机体的抗肿瘤免疫功能有关。

Abstract:

[Objective] This paper was designed to investigatethe anti-tumor effect of Fufang Huangqi Oral Solution (FFHQKY) combined chemotherapyin H22 tumor-bearing mice and reveal its mechanism. This study will provided a basis for the theoretical foundation of antitumor immunotherapy of FFHQKY.[Methods] The H22 tumor-bearing mice were established to investigate the effect ofanti-tumor immunological effect of FFHQKY combined chemotherapy. Mice were divided into four groups, including model group, FFHQKY treated-group, cyclophosphamide(CTX) group, and combination treated group. After 16 days of intragastric administration, the tumor tissue were isolated to assess the anti-tumor effect of FFHQKY, the lymphocyte cells proliferation was detected by MTT method, the total cell numbers of leukocyte, monocyte, neutrophil were measured by hematology analyzer. The expression of Treg (CD4⁺Foxp3⁺) and Th1 cells(CD4⁺IFN-γ⁺) were detected by flow cytometry.[Results] Compared with CTX group, the combination treated group had no influence on inhibitory rate of tumor but significantly recovered the lymphocyte proliferation activity from H22 tumor-bearing mice in response to ConA, increased the cells count of leukocyte and neutrophil. Importantly, compared with CTX group, the percent of Th1 cellsTh1 cells (CD4⁺IFN-γ⁺) were increased in combination treated group. Furthermore, combination treatment reduced the expression of Treg (CD4⁺Foxp3⁺).[Conclusion] Combination FFHQKY with chemotherapy exerted significantly anti-tumor immunological, the mechanism of action may be linked to activate Th1 cells response and down-regulated Treg expression, consequently enhance anti-tumor immunological effect.