【摘要】 目的 研究胀果甘草粗多糖(GiP)及其纯化多糖GiP-B1对荷瘤小鼠树突状细胞(DC)成熟和抗肿瘤作用的影响及机制。方法 将体外培养的肝癌细胞H22荷瘤小鼠未成熟DC(imDC)分为对照组、肿瘤坏死因子α(TNF-α)组、GiP组及GiP-B1组，检测荷瘤小鼠成熟DC(mDC)的细胞活力，表面标志物(CD11c、CD80、CD86、MHC-Ⅱ)阳性表达率，白细胞介素12p70(IL-12p70)及IL-4水平；通过荷瘤小鼠m DC与CD4~+T淋巴细胞共培养生成CD4-细胞毒性T细胞(CD4-CTL)，检测刺激指数，CD4-CTL上清液中IL-12p70、干扰素-γ(IFN-γ)、IL-4、IL-10水平及对H22细胞的杀伤活性；检测共培养后荷瘤小鼠mDC中IL-12、IL-12受体(IL-12R)、信号转导和转录激活因子4(STAT-4)mRNA表达量，以及IL-12Rβ2蛋白表达量，核转录因子κB(NF-κB)p65、STAT-4蛋白磷酸化水平。结果 与对照组比较，GiP组与GiP-B1组荷瘤小鼠mDC细胞活力、MHC-Ⅱ阳性表达率以及IL-12p70、IL-4水平均显著升高(P<0.05),CD11c、CD80、CD86阳性表达率均有升高趋势，但差异均无统计学意义(P>0.05)。共培养后刺激指数、IL-12p70、IFN-γ水平均显著升高(P<0.05),IL-4、IL-10(GiP组除外)水平均显著降低(P<0.05)，对H22细胞的杀伤活性均显著增强(P<0.05)；荷瘤小鼠m DC中IL-12、IL-12R(GiP组除外)、STAT-4 mRNA表达量，IL-12Rβ2蛋白表达量以及NF-κB p65、STAT-4蛋白磷酸化水平均显著升高(P<0.05)。结论 GiP及GiP-B1对荷瘤小鼠DC的成熟有较好的促进作用，能有效刺激CD4~+T细胞增殖并增强CD4-CTL的抗肿瘤活性，其作用机制可能与激活IL-12/NF-κB/STAT-4信号通路有关。更多还原

【Abstract】 OBJECTIVE To investigate the effects and mechanism of Glycyrrhiza inflata polysaccharides(GiP) and GiP-B1 on the maturation and anti-tumor effect of dendritic cell(DC). METHODS The immature DC(imDC) of hepatocellular carcinoma cell H22 tumor-bearing mice cultured in vitro were divided into control group, tumor necrosis factor-α(TNF-α) group, GiP group, and GiP-B1 group. The viability, positive expressions of surface markers(CD11c, CD80, CD86, MHC-Ⅱ), the levels of interleukin-12p70(IL-12p70) and IL-4 in mature DC(m DC) of tumor-bearing mice were detected. mDC and CD4~+T lymphocytes were co-cultured to generate CD4-cytotoxic T cell(CD4-CTL); stimulation index, the levels of IL-12p70, interferon-γ(IFN-γ), IL-4 and IL-10, the killing activity of CD4-CTL to H22 cell were detected. mRNA expressions of IL-12, IL-12 receptor(IL-12R), signal transducer and activator of transcription-4(STAT-4), as well as the protein expression of IL-12 receptor β2(IL-12Rβ2), phosphorylation levels of nuclear factor-kappa B(NF-κB) p65 and STAT-4 proteins in mDC were detected after co-culture.RESULTS Compared with the control group, the viability of mDC, the positive expressions of MHC-Ⅱ, and the levels of IL-12p70 and IL-4 were increased significantly in GiP group and GiP-B1 group(P<0.05). The positive expressions of CD11c, CD80 and CD86 showed an increasing trend, but the differences were not statistically significant(P>0.05). After co-culturing, the stimulation index, the levels of IL-12p70 and IFN-γ were significantly increased(P<0.05), while the levels of IL-4 and IL-10(except for the GiP group) were significantly decreased(P<0.05); the cytotoxicity against H22 cells was significantly enhanced(P<0.05). mRNA expressions of IL-12 and IL-12R(except for GiP group) and STAT-4, protein expression of IL-12Rβ2 as well as phosphorylation levels of NF-κB p65 and STAT-4 protein were increased significantly in mDC(P<0.05). CONCLUSIONS GiP and GiP-B1 have a good promoting effect on the maturation of DC in tumor-bearing mice, effectively stimulate CD4~+T cell proliferation, enhance the anti-tumor activity of CD4-CTL, and its mechanism may be related to activating IL-12/NF-κB/STAT-4 signaling pathway.更多还原