紫草素对人外周血单核细胞来源的树突状细胞表型及功能的影响

The effect of Shikonin on monocyte-derived DCs phenotype and function

目的探讨紫草素对人外周血单核细胞来源的树突状细胞表型及功能的影响。方法从健康人外周血中分离获得单个核细胞,经过夜贴壁后获得单核细胞,体外采用多种细胞因子联合诱导,获得了分化与功能相对成熟的树突状细胞。采用流式细胞仪检测技术观察不同质量浓度紫草素对DC表面分子表达的影响,采用CCK-8法观察紫草素对树突状细胞促淋巴细胞增殖以及ELISA法检测其对树突状细胞分泌细胞因子IL-23的干预作用。结果 TNF-α25 ng/ml刺激树突状细胞,细胞表面分子CD80、CD83表达升高,刺激同种异体淋巴细胞增殖能力增强;100μg/ml紫草素可抑制CD80及CD86的表达,50μg/ml紫草素可抑制CD86的表达;100μg/ml紫草素和50μg/ml紫草素均可抑制树突状细胞促淋巴细胞增殖的能力;100μg/ml紫草素和50μg/ml紫草素均可抑制LPS和INF-γ联合诱导的树突状细胞IL-23的分泌。结论紫草素具有一定的抑制树突状细胞成熟及抑制其分泌IL-23的作用。

Dendritic cells（DC） are main gate-keepers of the immune system, bridging the innate and adaptive immune system. DCs are able to mature into inflammatory DCs at sites of inflammation in both autoimmune and allergic disease, such as psoriasis. Shikonin is a kind of blood-cooling and toxin-relieving herb, which is effective in psoriasis treatment. In this study, we attempt to investigate the effect of Shikonin on cell phenotype and function of monocyte-derived DCs. Firstly, peripheral blood mononuclear cell（PBMC） were purified from buffy coats of healthy donors and cultured overnight, then the adherent cells were selected and matured with several pro-inflammatory cytokines or combinations of pro-inflammatory cytokines. Flow cytometery was used to analyze the effect of Shikonin with different concentration on inflammatory DCs phenotype; the mixed lymphocyte reaction were tested by cell counting kit-8; IL-23 secretion were analyzed by ELISA kit. The data showed that TNF-α（25 ng/ml）induced a high expression of the activation markers CD80 and CD86; DCs treated with TNF-α（25 ng/ml） stimulated allogeneic T-cell proliferation; 100 μg/ml Shikonin suppressed the expression of CD80 and CD86, while 50 μg/ml Shikonin only suppressed the expression of CD86; both 100 μg/ml and 50 μg/ml Shikonin suppressed the ability of DCs on stimulating leukomonocyte and secreting IL-23. Our finding indicated that Shikonin can suppress the maturation and IL-23 secretion of DCs to some extent.