胀果甘草粗多糖及其纯化多糖对树突状细胞成熟和抗肿瘤作用的影响及机制
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篇名: 胀果甘草粗多糖及其纯化多糖对树突状细胞成熟和抗肿瘤作用的影响及机制
TITLE: Effects and mechanism of Glycyrrhiza inflata polysaccharides on the maturation and anti-tumor effect of dendritic cell
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mDC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mDC中IL-12、IL-12R(GiP组除外)、STAT-4mRNA表达量,IL-12Rβ2蛋白表达量以及NF-κBp65、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。
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 (mDC) 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.
期刊: 2024年第35卷第20期
作者: 娜迪热木·肖克拉提;俞永婷;卢泳强;周朝曦;张丽丽;丛媛媛
AUTHORS: Nadiremu·Xiaokelati ,YU Yongting,LU Yongqiang,ZHOU Zhaoxi,ZHANG Lili,CONG Yuanyuan
关键字: 胀果甘草多糖;树突状细胞;IL-12/NF-κB/STAT-4信号通路;细胞毒性T淋巴细胞;抗肿瘤
KEYWORDS: Glycyrrhiza inflata polysaccharides; dendritic
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