雷公藤免疫调节的体外研究

The study on immuno-modulation of triptolide in vitro

目的证实雷公藤能否选择性地抑制LPS、PHA诱导的TNFα、IFNγ的产生,以及它抑制TNFα的免疫学机理。方法采用健康供血者外周血单个核细胞(PBMC)的体外培养及ELISA,测量雷公藤甲素抑制由LPS、PHA诱导的TNFα、IFNγ的能力;通过对PBMC和单核细胞(monocyte)的预处理试验,以及其后的流式细胞仪分析,研究雷公藤抑制TNFα的免疫学机理。结果雷公藤对TNFα、IFNγ的抑制具有剂量依赖性,对TNFα的IC50为5～10ngml,对IFNγ的IC50为0.1～1.0ngml。PBMC和monocytes试验中,雷公藤的不同预处理后的TNFα浓度,与流式细胞分析产生效应的CD14+TNFα+细胞数一致。结论两种预处理方法提示了不同的临床意义。免疫表现型的分析揭示,雷公藤可能与LPS竞争结合CD14受体。该研究为雷公藤可作为抗炎剂治疗麻风反应提供了基本依据。

Objective To find whether triptolide can inhibit selectively TNF-α and its mechanism. Methods The ability of triptolide to inhibit the production of TNF-α and IFN-γ stimulated by LPS and PHA by PBMC from healthy donors was measured by ELISA assay. The immunological mechanism of triptolide on TNF-α was investigated by pre-treatment with triptolide of PBMC and monocytes followed by analysis with Flow cytometry (FCM). Results The inhibition of TNF-α and IFN-γ by triptolide occurred in a dose dependent manner and the CI_~50 was equal to 5-10ng/ml for TNF-α and 0.1-1.0ng/ml for IFN-γ. The concentrations of TNF-α measured after the different pre-treatments with triptolide on PBMC and monocytes are consistent with its effects on a population of CD14+/TNF-α+ monocytes shown on FCM. Conclusion Triptolide may compete with LPS for binding to the CD14 receptorand triptolide may be used as an anti-inflammatory reagent for the treatment of leprosy.