参山颗粒对CT26.WT荷瘤小鼠的免疫调节及抗肿瘤作用

Immunomodulatory and anti-tumor effects of Shenshan Granules on CT26.WT tumor-bearing mice

目的探讨参山颗粒对CT26.WT结肠癌荷瘤小鼠的免疫调节及抗肿瘤作用,为其功能定位、临床运用与市场推广提供理论依据及实验支撑。方法采用腋下注射CT26.WT细胞制备结肠癌小鼠模型,分为空白对照组(Control组)、模型组(Model组)、参山颗粒组(SSG组)、5氟尿嘧啶阳性药组(5-FU组),每组6只,连续给药14 d。计算肿瘤体积、肿瘤抑制率。血清中细胞因子中IL-2、TNF-α和IFN-γ的表达水平、脾脏中T淋巴细胞亚群的占比、肿瘤组的病理变化及肿瘤中Bcl-2、Bax和Caspase-3 mRNA和蛋白的表达情况,分别用ELISA检测法、流式细胞术、HE染色、RT-PCR法和免疫组化测定。结果与Model组对比,5-FU组和SSG组肿瘤体积显著减小、肿瘤质量显著降低、抑瘤率显著增加(P<0.01),5-FU组、血清中细胞因子IL-2、TNF-α和IFN-γ水平、脾细胞悬液中CD4^(+)T/CD8^(+)T的比值及肿瘤组织中Bcl-2 mRNA和蛋白表达显著降低(P<0.01),Bax和Caspase-3 mRNA和蛋白表达显著增加(P<0.01);SSG组血清中细胞因子IL-2、TNF-α、IFN-γ水平,脾细胞悬液中CD4^(+)T/CD8^(+)T的比值及肿瘤组织中Bax和Caspase-3 mRNA和蛋白表达显著增加(P<0.01),Bcl-2 mRNA和蛋白表达显著降低(P<0.01)。结论参山颗粒可能通过增强荷瘤小鼠免疫抗肿瘤,激活Bcl-2/Bax/Caspase-3细胞凋亡通路从而抑制CT26.WT结肠癌荷瘤小鼠肿瘤生长。

Objective To explore the immunomodulatory and anti-tumor effects of Shenshan Granules on CT26.WT colon tumor-bearing mice,providing theoretical basis and experimental support for its function orientation,clinical application,and market promotion.Methods The mouse model of colon cancer was prepared by subaxillary injection of CT26.WT cells.Then the mice were divided into blank control group(control group),model group,Shenshan Granules group(SSG group),and 5-fluorouracil positive drug group(5-FU group),with 6 mice in each group.The mice were administered drug continuously for 14 days.The tumor volume and tumor inhibition rate were calculated.The expression levels of IL-2,TNF-α,and IFN-γin the serum cytokines,the proportion of T lymphocyte subsets in the spleen,the pathological changes in the tumor group,and the mRNA and protein expressions of Bcl-2,Bax,caspase-3 in tumors were determined by ELISA,flow cytometry,HE staining,RT-PCR,and immunohistochemistry,respectively.Results Compared with the model group,the 5-FU and SSG groups had significantly smaller tumor volumes,lower tumor mass,and higher tumor inhibition rates(P<0.01).In the 5-FU group,IL-2,TNF-α,and IFN-γlevels in serum cytokines,the ratio of CD4^(+)T/CD8^(+)T in splenocyte suspensions,and the mRNA and protein expressions of Bcl-2 in tumor tissue significantly decreased(P<0.01).In the SSG group,IL-2,TNF-α,and IFN-γlevels in serum cytokines,the ratio of CD4^(+)T/CD8^(+)T in splenocyte suspensions,and the mRNA and protein expressions of Bax and caspase-3 in tumor tissue significantly increased(P<0.01),but the mRNA and protein expressions of Bcl-2 significantly decreased(P<0.01).Conclusion Shenshan Granules may inhibit tumor growth in CT26.WT colon tumor-bearing mice by enhancing the anti-tumor immunity and activating the Bcl-2/Bax/Caspase-3 apoptosis pathway.