[Mechanism of Shuangshen Fuzheng Powder in regulating TAMs towards M1 polarization to inhibit lung cancer].

This study aims to explore the inhibitory mechanism of Shuangshen Fuzheng Powder(SSG) on the tumor of mice bearing M1 macrophages and Lewis lung cancer cells. Lewis lung cancer cells and polarized RAW264.7 macrophages were mixed for injection to construct a mouse model with lung cancer xenograft. The mice were divided into a Lewis group, an SSG group, a Lewis + M1 macrophage(M1) group, and a Lewis + M1 + SSG group, with 10 mice in each group. The SSG group and the Lewis + M1 + SSG group were administered SSG suspension(208 mg·mL~(-1)) daily by gavage, and the Lewis group and the Lewis + M1 group were given purified water by gavage. The spleen and tumor samples were collected after three weeks of intervention. The volume and weight of subcutaneously transplanted tumors in mice were detected. The proportion of F4/80~+CD86~+ tumor-associated macrophages(TAMs) and the expression of CD206~+ in spleen cells were analyzed by flow cytometry, and the expression of M1 macrophage marker protein inducible nitric oxide synthase(iNOS) in transplanted tumor tissue was detected by immunohistochemical staining(IHC). The expression of iNOS, arginase 1(ARG1), transforming growth factor-β(TGF-β), vascular endothelial growth factor(VEGF), matrix metalloproteinase-9(MMP-9), and macrophage migration inhibition factor(MIF) in transplanted tumor tissue of mice was detected by Western blot(WB), and the chemical constituents of SSG were analyzed by ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry(UHPLC-Q-Exactive Orbitrap HR-MS). The results showed that treatment by SSG and co-injection with M1 macrophages could both significantly inhibit the tumor volume and weight, and the inhibitory effects were similar. During treatment by SSG, the tumor burden in the Lewis + M1 + SSG group continued to decrease. The flow cytometry results showed that the SSG group and the Lewis + M1 + SSG group exhibited a significant increase in the CD86~+ expression and a significant decrease in the CD206~+ expression. The IHC results indicated that compared with that of the Lewis group, the expression of iNOS in the SSG group was significantly elevated. The WB assay results showed that the expression of iNOS protein significantly increased, and the expression of ARG1, TGF-β, VEGF, MMP-9, and MIF significantly decreased in the Lewis + SSG group compared with those of the Lewis group. A total of 125 chemical constituents were identified from SSG, mainly including saponins, flavonoids, nucleotides, amino acids, and sugar components. SSG may exert its inhibitory effect on lung cancer by increasing the polarization of macrophages to M1-type TAMs and reducing the polarization to M2-type TAMs. The analysis of the pharmacodynamic components of SSG provides a basis and reference for subsequent research on pharmacokinetics.