The Mechanism Of Total Saponins Of Panax Ginseng Mediated NK Cell Activity In Promoting The Anticancer Activity Of 5-Fu

Objective:To explore the inhibitory effects of Total saponins of Panax ginseng(TSPG)combined with 5-fluorouracil(5-Fu)on orthotopic liver transplantation tumor mice and its mechanisms.Methods:1.The orthotopic transplantation tumor models were established and randomly divided into four groups:model group,TSPG group,5-Fu group and TSPG+5-Fu group.2.The tumor weight,tumor volume,tumor inhibitory rate,liver index and speen index were determined.3.The pathological changes in orthotopic liver transplantation tumor mices were surveyed with HE staining.4.The detection of NK1.1~+cells in liver tumor tissue and the protein expression levels of HIF-1α、VEGF、VEGFR-2 were observed by immunohistochemical.5.Flow cytometry was performed to determin the proportion of NK cells,CD4~+T cells and CD8~+T cells in liver and spleen.6.The expressions of perforin and granzyme B mRNA in liver tissues of orthotopic transplantation tumor mices were detected by QRT-PCR.Results:1.The effect of the TSPG on tumor growth in orthotopic liver transplantation tumor mice.Compared with the model group,tumor volumes,tumor weights and tumor inhibitory rate in other groups were significantly lower(P<0.05).2.The effect of the TSPG on liver index and liver index in orthotopic liver transplantation tumor mice.There is no significant difference in liver index between model group and TSPG group.Compared with the model group,spleen index in TSPG group increased significantly(P<0.05).3.The effect of the TSPG on pathological changes on liver in orthotopic liver transplantation tumor mice.The results of HE staining showed that the tumor cells in the model group were densely arranged and infiltrated and the cell heteromorphity was significant,and a large number of inflammatory cells were found in the tumor infiltrating necrosis area of the TSPG group of the tumor bearing mice,and the liver tissue structure of the5-Fu group of the tumor bearing mice was in disorder and appeared vacuoles,and the inflammatory cells were infiltrated around the tumor cells,and the vacuolated changes of the parenchymal cells in the group TSPG+5-Fu were less than those in the 5-Fu group.Obvious necrosis and infiltration of inflammatory cells were found.4.The effect of TSPG on NK1.1~+cell infiltration in orthotopic liver transplantation tumor mice.The immunohistochemical results showed that,compared with the model group,the infiltration degree of NK1.1+cells in the TSPG group increased significantly in the liver tumor tissues,mainly in the para-cancerous tissues and in the tumor necrosis area.5.Effects of TSPG on HIF-1,VEGF and VEGFR-2 in liver tumor tissues of orthotopic liver transplantation tumor mice.Immunohistochemical results showed that compared with the model group,the protein expression of HIF-1,VEGF and VEGFR-2 increased significantly in group TSPG(P<0.05).6.Effects of TSPG on NK cells and T cell subsets in liver and spleen in orthotopic liver transplantation tumor mice.The results of flow cytometry showed that compared with the model group,the proportion of liver and spleen NK cells in the liver and spleen in the TSPG group increased significantly(P<0.01),and the proportion of CD4+T cells and CD8+T cells in the TSPG group was also significantly increased(P<0.05).Compared with the 5-Fu group,the proportion NK cells,CD4~+T and CD8~+T cells in the spleen and liver in the TSPG+5-Fu group increased significantly(P<0.05).7.Effect of TSPG on the expression of perforin and granzyme B mRNA in orthotopic liver transplantation tumor mice.QRT-PCR results showed that compared with the model group,the expression of perforin and granzyme B mRNA in TSPG group and TSPG+5-Fu increased significantly(P<0.05).Compared with the 5-Fu group,the expression of perforin and particulate B mRNA was increased in TSPG+5-Fu group(P<0.05).Conclusion:1.TSPG could significantly reverse the inhibitory effect of 5-Fu on NK cells,CD4~+T cells and CD8~+T cells in orthotopic liver transplantation tumor mice.2.TSPG may increase the killing activity of NK cells by regulating the secretion of perforin and granzyme,thus enhancing the inhibitory effect of 5-Fu on the growth of mouse orthotopic liver transplantation tumor.