目的 观察牛膝有效部位在肿瘤微环境下对树突状细胞成熟的促进及功能调控作用。方法 干燥牛膝粉末提取并鉴定牛膝总皂苷和牛膝多糖;建立模拟肿瘤微环境的MCF-7乳腺癌细胞与树突状细胞共培养体系,并分别给予牛膝多糖、牛膝总皂苷及牛膝多糖和总皂苷协同刺激;流式细胞仪检测树突状细胞表面分子的表达及树突状细胞诱导的细胞毒性T淋巴细胞(CTL)杀伤活性,ELISA法检测培养上清中白细胞介素-1β、肿瘤坏死因子-α、白细胞介素-10和白细胞介素-12的水平,Western blot检测JAK/STAT3信号通路中p-JAK、JAK、p-STAT3、STAT3的蛋白表达。结果 在肿瘤微环境下,牛膝有效部位牛膝总皂苷和牛膝多糖均可以通过TLR4及Dectin-1受体途径,启动树突状细胞细胞JAK/STAT3信号通路,上调树突状细胞细胞表面分子CD80、CD86及CD40的表达,从而诱导树突状细胞的成熟与分化,促进肿瘤坏死因子-α、白细胞介素-12、白细胞介素-10及白细胞介素-1β等细胞因子的分泌,两者协同较单独应用作用加强。结论 以适当形式的中药有效部位作为免疫增强剂联合树突状细胞疫苗, 可有效纠正肿瘤微环境下的免疫缺陷,有效激活肿瘤特异性CTL细胞,从而产生特异性抗肿瘤效应。
Abstract
OBJECTIVE To study the active part of Achyranthes bidentata promotes maturation and regulates the functions of DC in tumor microenvironment. METHODS The total saponins and the polysaccharides from dry powder of Achyranthes bidentata were extracted and identified. The tumor microenvironment by co-culturing breast cancer MCF-7 cells with DCs, and add polysaccharides, total saponins or total saponins with polysaccharides of Achyranthes bidentata to stimulate DCs were established. The expression of surface molecules on DCs and the killing activity of cytotoxic T lymphocytes (CTL) induced by DCs by Flow Cytometry were detected. ELISA was used to detect the levels of IL-1β, TNF-α, IL-10 and IL-12 in the culture supernatant. Detect the protein expression of JAK, p-JAK, STAT and p-STAT3 in DCs by Western blot. RESULTS In the tumor microenvironment, the active parts of Achyranthes bidentata, total saponins and ABPS, alone or combined used, can trigger JAK/STAT3 signaling pathway of DCs through TLR4 and Dectin-1 receptor and upregulate the expression of surface molecules, such as CD80, CD86 and CD40, to induce the maturation and differentiation of DCs, promote the secretion of TNF-α, IL-12, IL-10 and IL-1β. CONCLUSION As an effective adjuvant, the active parts of traditional Chinese medicine can enhance the effect of DC vaccine to correct the immune deficiency in tumor microenvironment, activate tumor-specific CTLs and then produce specific anti-tumor effects.
关键词
牛膝 /
树突状细胞 /
肿瘤微环境
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Key words
Achyranthes bidentata /
dendritic cell /
tumor microenvironment
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中图分类号:
R965
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参考文献
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脚注
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基金
辽宁省教育厅科学研究一般项目资助项目(L2012331)
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