Effect Of Astragalus Polysaccharides (APS) On The Dendritic Cells Cultured In The Supernatant Of HepG2 Cells And HepG2.2.15 Cells

Objective To study the effect of APS on the dendritic cells cultured in the supernatant of HepG2 cells and HepG2.2.15 cells and investigate the mechanism of hepatitis B-related hepatic carcinoma from the points of antigen presenting cells and immune escape to provide new ideas for prevention and treatment of hepatic carcinoma.Methods This experiment used the density-grads centrifugal method to apart the peripheral blood monocytes, and human dendritic cells were induced in vitro by the application of tumor necrosis factor-a (TNF-a), interleukin-4 (IL-4) and GM-CSF. We received the conditioned meduim by gathering two different hepatica carcinoma cell supernatant (HepG2, HepG2.2.15). This experiment was divided into two subgroups: the control group and the drug group. The morphology of DCs was identified by inverted microscope and the phenotype (HLADR, CD86, CDla, and CD83) of cultured 7 days DCs was identified by flow cytometry. The capability of stimulating allo-lymphocyte proliferation was tested with mixed leukocyte reaction (MLR) by MTT and the expressions of IL-12p70 and IFN-y in the supernatant of DCs and MLR were detected by enzyme linked immunosorbent assay (ELISA). The NF-κB expressions in nucleus were detected by ELISA.Results1. A large number of functional DCs could be induced from the healthy human peripheral blood monocytes by the application of tumor necrosis factor-a (TNF-a), interleukin-4 (IL-4) and GM-CSF.2. In the control group, the phenotype (HLADR, CD86, CD1a, CD83), capability of stimulating allo-lymphocyte proliferation, and expressions of IL-12p70, IFN-γNF-κB in nucleus of DCs cultured in complete medium and conditioned medium were lower than those of DCs in the drug group(P<0.05).3. In the control group and drug group, the phenotype (HLADR. CD86, CDla, CD83), capability of stimulating allo-lymphocyte proliferation, and expressions of IL-12p70, IFN-y, NF-κB in nucleus of DCs cultured in complete medium were higher than those of DCs cultured in conditioned medium (P<0.05).4. In the control group and drug group, the phenotype (HLADR, CD86, CD la, CD83), capability of stimulating allo-lymphocyte proliferation, the expressions of IL-12p70, IFN-y, and NF-κB in nucleus of DCs cultured in HepG2 CM were higher than those of DCs cultured in 25%HepG2.2.15 CM (P<0.05).Conclusions1.Conditioned medium from hepatica carcinoma cell supernatant can cause functional defect of DCs, inhibit the phenotype and expressions of IL-12p70, IFN-y, NF-κB in nucleus, and affect the differentiation and maturation of DCs, suggesting that its dysfunction may be associated with the inhibition of NF-κB signaling pathway.2. APS can raise the phenotype, IL-12p70 expressions and IFN-γexpressions of DCs from healthy human peripheral blood and DCs cultured in conditioned medium. APS can promote the DCs differentiation and maturation in the normal environment, tumor microenvironment or virus microenvironment and also enhance immune function.3. APS can raise the NF-κB expressions of DCs in normal environment and DCs with functional defect in virus microenvironment, suggesting that the mechanism of APS promoting DCs maturation may be related to NF-κB signaling pathway.