基于肿瘤微环境探讨六神丸对结肠癌小鼠移植瘤的抗肿瘤作用

Antitumor Effect of Liu-Shen-Wan on Transplanted Tumors of Mice with Colon Cancer from Perspective of Tumor Microenvironment

目的基于肿瘤微环境中的M2型巨噬细胞极化探讨六神丸对结肠癌小鼠移植瘤的作用。方法建立CT26结肠癌小鼠皮下移植瘤模型,将小鼠随机分为对照组、奥沙利铂组、奥沙利铂联合六神丸组,给药过程中测量肿瘤体积及小鼠体质量。三周后处死小鼠,称取瘤重,计算脾脏指数,同时检测血清中IFN-γ、IL-12P40表达水平及相关血生化指标。进一步检测血清及肿瘤组织中IL-10与TGF-β的表达水平,免疫组织化学实验检测各组M2型巨噬细胞浸润程度。结果与对照组相比,奥沙利铂联合六神丸组小鼠的肿瘤体积及体质量明显下降,小鼠的脾脏指数及血清中IFN-γ、IL-12P40表达水平明显增高,且无明显不良反应。IL-10与TGF-β的表达水平明显下降,CD68及CD206表达水平下降。结论六神丸对结肠癌小鼠移植瘤的抗肿瘤作用与抑制肿瘤微环境中M2型巨噬细胞极化密切相关。

Objective To investigate the effect of Liu-Shen-Wan on transplanted tumors in mice with colon cancer based on the polarization of M2 macrophages in the tumor microenvironment.Methods We established a subcutaneous transplantation tumor model of mice with CT26 colon cancer.Mice were randomly divided into vehicle,oxaliplatin,and oxaliplatin combined with Liu-Shen-Wan groups.Treatment was administered for three weeks,and tumor volume was measured.All mice were weighed during the administration.After the end of the treatment,the mice were dissected and tumors were photographed and weighed.Spleen index was calculated.The expression levels of IFN-γand IL-12P40 in serum and related blood biochemical indices were measured.The expression levels of M2 macrophage polarization indices,namely,IL-10 and TGF-β,in serum and tumor tissues were detected.The infiltration degree of M2 macrophages in each group was observed by immunohistochemical experiments.Results The tumor volume and mouse weight in the oxaliplatin combined with Liu-Shen-Wan group significantly decreased compared with those in the vehicle group.The spleen index increased,and the expression levels of IFN-γand IL-12P40 in serum also significantly increased.The mice had no obvious side effects after the drug treatment.In addition,the expression levels of IL-10 and TGF-βin the serum and tissues of mice in the oxaliplatin combined with Liu-Shen-Wan group significantly decreased.The expression levels of CD68 and CD206 in tumor tissues also decreased.Conclusion The anti-tumor effect of Liu-Shen-Wan on the transplanted tumors of mice with colon cancer is related to the inhibition of M2 macrophage polarization in the tumor microenvironment.