刺山柑果实总生物碱粗提物对小鼠树突状细胞成熟的影响

Effects of crude extract of Capparis spinosa L. fruit alkaloids on the maturation of murine dendritic cells

目的探讨刺山柑(Capparis spinosaL.)果实总生物碱粗提物对小鼠骨髓来源树突状细胞(dendritic cell,DC)成熟的影响。方法提取刺山柑果实总生物碱粗提物(Capparis spinosa fruit alkaloids,CSFA),高效液相色谱对其含量进行测定。用不同浓度(1、2、3mg/ml)的CSFA处理DC,采用流式细胞术检测DC活性、表面分子表达、抗原吞噬能力,ELISA检测细胞因子分泌。Westernblot检测MAPK及NF-κB信号通路关键分子的活化水平。结果CSFA单独处理时对DC细胞因子及共刺激分子的表达无显著影响,然而,与细菌脂多糖(lipopolysaccharides,LPS)联用时显著降低了LPS诱导的CD40、CD80、CD86及MHCⅡ的表达,IL-12p40及TNF-α的分泌,但是增加了IL-10的表达及DC抗原吞噬能力。进一步研究表明,CSFA降低了LPS诱导的p38、ERK、JNK的磷酸化及核内NF-κBp65的水平。结论CSFA可能通过MAPK及NF-κB信号通路抑制了LPS诱导的DC成熟及促炎细胞因子的表达,增加了抗炎细胞因子IL-10的表达及抗原吞噬能力,说明CSFA具有潜在的免疫抑制作用。

Objective To investigate the effects of crude extract of Capparis spinosa L. fruit alkaloids (CSFA) on the maturation of murine bone marrow-derived dendritic cells (DCs). Methods CSFA was prepared and the contents were determined by high performance liquid chromatography. DCs were treated with different doses (1, 2, 3 mg/ml) of CSFA. The viability of DCs, the expression of surface molecules and the ability of phagocytosis were detected by flow cytometry. The secretion of cytokines was measured by ELISA. Western blot assay was performed to analyze the activation of key molecules in mitogen-activated protein kinases (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways. Results The results showed that CSFA alone had no significant influence on the expression of surface molecules and cytokines in DCs. However, it significantly decreased the expression of CD40, CD80, CD86 and MHC Ⅱ as well as the secretion of IL-12p40 and TNF-α that were induced by lipopolysaccharides (LPS), but increased IL-10 secretion and the ability of phagocytosis after treating DCs with both CSFA and LPS. Further, the phosphorylation of p38, extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) and the nuclear translocation of NF-κBp65 induced by LPS were inhibited by CSFA. Conclusion CSFA could inhibit the maturation of DCs and the secretion of pro-inflammatory cytokines induced by LPS while increasing the secretion of the anti-inflammatory cytokine IL-10 and the ability of phagocytosis, which might involve MAPK and NF-κB signaling pathways. This study suggests that CSFA could be used as a potential immunosuppressant.