Mice On The Centralizer Detoxification Party Before Gastric Cancer Induced By Tumor Associated Macrophage Of Intervention Study

Background:Relapse and metastasis of malignant tumor have always been the key problem that restricts the clinical efficacy in cancer treatment. In cancer patients after resection of primary tumor, some immune regulating cells and cytokines have close relation with tumor relapse and metastasis. In the past, researchers thought that macrophages played an important role in anti-cancer immune regulation in that macrophages could directly kill cancer cell, or eliminate cancer cells by presenting certain tumor antigen to activate body immune response. However, as more researches are devoted to tumor stroma and cancer inflammation, it comes to realized that tumor-associated macrophages (TAMs) have the property of M2-polarized macrophages and play a role in promoting cancer development, invasion, metastasis, suppressing immune response and promoting tumor neovascularization and microlymphatic formation rather than anti-cancer effect. Fu Zheng Jie Du (FZJD) method is the major principle to prevent cancer metastasis after surgery in TCM cancer treatment. Possible mechanism may be enhancing phagocytosis of intraperitoneal macrophages, enhancing immune surveillance and inhibiting immune escape of cancer cells. Based on the modern researches of TAM and TCM clinical experiences, we propose that Fu Zheng Jie Du formula may prevent cancer metastasis by regulating M2tumor-associated macrophages to prevent tumor metastasis.Methods:60615mice were randomly divided into5groups,12mice in each group: blank control group (B), model control group (C), Fu Zheng Jie Du group (FZJD),5-Fu group (5-Fu),5-Fu+FZJD group (5-Fu+FZJD). Mice in blank control group were non-tumor-bearing models, while the other four groups were tumor-bearing models (injected with MFC cells). Mice in model control group were administrated by gavage with distill water for21days. Mice in FZJD group were administrated by gavage with FZJD decoction for21days. Mice in5-Fu group were administrated by intraperitoneal injection with5-Fu for7days. Mice in5-Fu+FZJD group were administrated by both intraperitoneal injection with5-Fu and gavage with FZJD decoction. Tumor diameters were measured every three days and tumor growth curves were drawn; On day21, weight the body weight of mice; lung tissues were removed and the inhibition rate of pulmonary metastasis were calculated; another 10mice for each group were used to observe the survival time; tumors were removed and tumor inhibition rates were calculated; the infiltration of CD68macrophages in tumor tissues were detected by immunohistochemistry; M2(F4/80CD206) macrophages were detected by flow cytometry; spleen CDllb F4/80macrophages cells and myeloid-derived suppressor cells (MDSC) were detected by flow cytometry; IL-4,11-10, IL-13and TGF-β in mice serum were detected by ELISA.Result:1. Tumor volumes:C> FZJD>5-Fu>5-Fu+FZJD,5-Fu+FZJD group had the lowest tumor grow rate, tumor volumes of5-Fu group and5-Fu+FZJD group were significantly smaller than model group (P<0.05);2. Tumor weight:C> FZJD>5-Fu>5-Fu+FZJD, model group had a maximum tumor weight of4.03±1.53g while5-Fu+FZJD had a minimum tumor weight of1.88±0.75g, the tumor weight of5-Fu group and5-Fu+FZJD group were significantly smaller than model group (P<0.05);3. Tumor inhibition rate:5-Fu+FZJD>5-Fu> FZJD,5-Fu+FZJD had the highest inhibition rate of53.35%;4. Lung metastasis:C> FZJD>5-Fu>5-Fu+FZJD,5-Fu+FZJD group had the highest metastasis inhibition rate of55.56% and was significantly higher than FZJD group (2.78%)(P<0.05);5. Survival time:C> FZJD>5-Fu>5-Fu+FZJD,mice in5-Fu+FZJD group lived the longest time;6. M2macrophages (F4/80+CD206+):M> FZJD>5-Fu>5-Fu+FZJD;7. Spleen macrophages (CDllb+F4/80+):M>5-Fu>FZJD>5-Fu+FZJD>C, compared to model group and5-Fu group, CDllb+F4/80+spleen macrophages in5-Fu+FZJD group were significantly lower (P<0.05);8. Myeloid immune suppressor cells MDSC (CDllb+Gr-1+): M>5-Fu>FZJD>5-Fu+FZJD>C, compared to model group, MDSC5-Fu+FZJD group and FZJD group were significantly lower (P<0.05),9. M2macrophage-induced cytokine: IL-4:M>5-Fu>FZJD>5-Fu+FZJD>C, TL-10:M> FZJD>5-Fu>5-Fu+FZJD> C, IL-13: M>FZJD>5-Fu>5-Fu+FZJD, TGF-β:M>5-Fu>FZJD>5-Fu+FZJD.Conclusion:Fu Zheng Jie Du decoction had a better effect in controlling tumor size,reducing lung metastasis of tumor-bearing mice, and improving the prognosis of tumor-bearing mice. The mechanism was to reduce CD68macrophage infiltration in tumor tissue, reduce M2macrophages induced cytokine IL-4, IL-10, IL-13, TGF-β, and protect the immune function by reducing immunosuppressive cells (CDllb F4/80macrophages, MDSC) in spleen.