中国中药杂志2025，Vol.50Issue(18)：5005-5015,11.DOI:10.19540/j.cnki.cjcmm.20250510.408
附子多糖联合仑伐替尼通过促进肿瘤相关巨噬细胞M1极化增强抗肝细胞癌作用研究
Study on enhanced anti-hepatocellular carcinoma effect of Fuzi polysaccharide combined with lenvatinib through promoting M1 polarization of tumor-associated macrophages
摘要
Abstract
To investigate the role of Fuzi polysaccharide(FPS)in combination with lenvatinib(LEN)in regulating tumor-associated macrophage(TAM)polarization to enhance the anti-hepatocellular carcinoma(HCC)efficacy,the non-toxic concentrations of FPS and LEN on RAW264.7 cells were screened by cell counting kit-8(CCK-8).The co-culture system of RAW264.7 and Hepa1-6 cells was established,and flow cytometry was used to detect changes in M1 and M2 macrophage phenotypes.Quantitative real-time polymerase chain reaction(qRT-PCR)was performed to assess the mRNA expression levels of M1-associated factors,inducible nitric oxide synthase(iNos)and interleukin 12b(IL-12b),as well as M2-associated factors arginase 1(Arg-1)and IL-10.The effects of macrophages treated with FPS and LEN on HCC cell proliferation,migration,invasion,and apoptosis were evaluated by 5-ethynyl-2'-deoxyuridine(EdU)staining,wound healing assay,Transwell invasion assay,and Annexin V/PI double-staining flow cytometry.A C57BL/6 subcutaneous HCC xenograft mouse model was established to observe the tumor growth,body weight,spleen index,and tumor-infiltrating macrophage polarization.CCK-8 assay showed that 50-200 mg·L-1 FPS and 1-5 μmol·L-1 LEN had no inhibitory effects on RAW264.7 macrophages.Flow cytometry and qRT-PCR results demonstrated that 200 mg·L-1 FPS+LEN significantly increased the proportion of M1 macrophages(P＜0.01)and the M1/M2 ratio(P＜0.01),upregulated the expressions of M1-associated genes iNos and IL-12b(P＜0.01),reduced the proportion of M2 macrophages(P＜0.01),and inhibited the expressions of M2-associated genes Arg-1 and IL-10(P＜0.01).Functional assays revealed that macrophages of the FPS+LEN group inhibited Hepa1-6 proliferation(P＜0.01),promoted apoptosis(P＜0.01),and reduced migration and invasion(P＜0.01).In vivo experiments showed that compared to those of the LEN group,the tumor volume was significantly reduced(P＜0.05),and the spleen index was significantly increased(P＜0.05)in the FPS+LEN group.The body weight was not decreased,and the proportion of M1 macrophages and the M1/M2 ratio in the tumors were significantly increased in the FPS+LEN group(P＜0.01).In conclusion,FPS combined with LEN enhances the anti-HCC effect by promoting the M1 polarization of TAMs.关键词
附子多糖/仑伐替尼/巨噬细胞/肝细胞癌Key words
Fuzi polysaccharide/lenvatinib/macrophages/hepatocellular carcinoma引用本文复制引用
卢林竹,黄振,郭倩倩,谭年花,吕长军,易纯,田雪飞..附子多糖联合仑伐替尼通过促进肿瘤相关巨噬细胞M1极化增强抗肝细胞癌作用研究[J].中国中药杂志,2025,50(18):5005-5015,11.基金项目
湖南省自然科学基金项目(2025JJ90042) （2025JJ90042）
湖南省教育厅科学研究项目优秀青年项目(23B0394) （23B0394）
湖南省自然科学基金青年基金项目(2023JJ40487) （2023JJ40487）
中国博士后科学基金项目(2022M721127) （2022M721127）
湖南省研究生科研创新项目(CX20230820) （CX20230820）
