双参颗粒对肺癌小鼠肿瘤相关巨噬细胞的抑制作用

Inhibitory effects of Shuangshen Granules on tumor-associated macrophages in mice with lung cancer

目的:验证双参颗粒抑制肺癌的有效性并探讨其机制。方法:构建荷Lewis肺癌小鼠模型,分为双参颗粒组、模型对照组,每组10只;构建KrasG12D自发肺癌小鼠模型,分为双参颗粒组、模型对照组,每组10只。分别给予双参颗粒、纯净水灌胃干预。观察荷Lewis肺癌小鼠的移植瘤质量,脾脏T淋巴细胞、肿瘤相关巨噬细胞(TAM)比例;KrasG12D自发肺癌小鼠肺组织micro-CT影像表现、肺组织病理改变、脾脏T淋巴细胞、TAM细胞比例,肺组织全转录组基因测序分析。结果:双参颗粒组移植瘤的瘤体质量显著低于模型对照组(P<0.01),双参颗粒组荷Lewis小鼠的脾脏CD_(3)^(+)CD_(4)^(+)T淋巴细胞比例与CD_(3)^(+)CD_(8)^(+)T淋巴细胞比例显著高于模型对照组(P<0.05),F4/80^(+)CD_(206)^(+)TAM细胞比例显著低于模型对照组(P<0.01)。模型对照组KrasG12D自发肺癌小鼠经micro-CT可以观察到肺部结节与占位,肺组织经HE染色进行病理学观察确定为肿瘤形成,与造模前正常小鼠肺组织比较,提示模型构建成功。双参颗粒组KrasG12D自发肺癌小鼠micro-CT扫描未观察到明确的肺部占位,经病理学观察无肿瘤形成。对两组KrasG12D自发肺癌小鼠进行全转录组测序分析提示,TAM相关的非编码基因miR-21a、miR-155、miR-146b表达存在差异。结论:双参颗粒能够抑制肺癌生长及形成,其作用机制可能与调节TAM细胞相关。

Objective:To verify the effectiveness of Shuangshen Granules(SSG)in inhibiting lung cancer and explore its mechanism.Methods:Mouse with bearing Lewis lung cancer cells(LLC)were divided into SSG group and control group,10 in each group.KrasG12 D spontaneous lung cancer mouse model was established and divided into SSG group and model group,10 in each group.SSG and purified water were administrated,respectilily.For LLC mouse,the quality of transplanted tumor,the proportion of spleen T lymphocytes and tumor associated macrophages(TAM)cells were observed.For KrasG12 D mouse,the micro-CT image of lung,the pathological of lung tissue,the proportion of spleen T lymphocytes and TAM cells were observed.Complete transcriptome sequencing analysis on lung tissues was conducted.Results:The weight of transplanted tumor in SSG group was significantly lower than that in control group(P<0.01),CD_(3)^(+)CD_(4)^(+)and CD_(3)^(+)CD_(8)^(+)T lymphocytes in spleen of LLC mouse in SSG group were higher than those in control group(P<0.05).The proportion of F4/80^(+)CD_(206)^(+)TAM cells was lower than that of control group(P<0.01).Tumor could be observed by micro-CT image in KrasG12 D mouse of control group,and pathological result supported this diagnosis.Pneumonia was observed by micro-CT image in KrasG12 D mouse in the SSG group.Pathological result supported this diagnosis.Complete transcriptome sequencing analysis of KrasG12 D mouse in both control group and SSG group suggested that the expressions of TAM related non-coding genes miR-21 a,miR-155 and miR-146 b were different.Conclusion:SSG can inhibit the lewis lung cancer growth and formation,and the mechanism of this effect may be related to TAM cells.