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资捷, 朱家勇. HPLC法测定人脑膜成纤维细胞IDO活性[J]. 中国公共卫生, 2003, 19(1): 53-54.
引用本文: 资捷, 朱家勇. HPLC法测定人脑膜成纤维细胞IDO活性[J]. 中国公共卫生, 2003, 19(1): 53-54.
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ZI Jie, ZHU Jia-yong. Determination on IDD activity of human meninx fibroblast with HPLC[J]. Chinese Journal of Public Health, 2003, 19(1): 53-54.
Citation: ZI Jie, ZHU Jia-yong. Determination on IDD activity of human meninx fibroblast with HPLC[J]. Chinese Journal of Public Health, 2003, 19(1): 53-54.
shu

HPLC法测定人脑膜成纤维细胞IDO活性

Determination on IDD activity of human meninx fibroblast with HPLC

    摘要
  • 摘要:
      目的   建立反相高效液相色谱法测定人脑膜成纤维细胞(Human meninx fibroblast, HMF)的吲哚胺2, 3-双加氧酶(indoleamine2, 3-dioxygenase, IDO)活性, 并探讨IDO的临床意义。
      方法   以荧光检测器和紫外检测器分别检测色氨酸和犬尿氨酸, 间接测定IDO活性, 采用3H-尿嘧啶(3H-U)特异标记的弓形虫在HMF内的增殖实验。
      结果   色氨酸和犬尿氨酸的平均回收率分别为9909%, 10218%, 变异系数分别为103%, 189%, 最低检测限分别为877, 3275ng/ml.被IFN-γ诱导产生的IDO可降解胞内必需氨基酸色氨酸而抑制弓形虫生长。
      结论   本法为一种灵敏、简便和可靠的反相高效液相色谱法, IDO可抑制弓形虫生长。

     

    Abstract:
      Objective   To determine indoleamine 2, 3-dioxygenase(IDO)activity in human meninx fibroblast(HMF)by using reversed-phase HPLC and establishing a simple and reliable method, and to explore clinical significance of IDO.
      Methods   IDO activity is indirectly determined by detecting the amount of tryptophan and kynurenine with FLD and DAD respectively.The ability to restrict growth of T.gondii was evaluted by 3H-U incroporation assay.
      Results   The average coefficients of recovery of tryptophan an kynurenine were 99.02% and 102.18% respectively, their coefficients of variation were 1.03% and 1.89% respectively, and their minimal detected concentration were 8.77 ng/ml and 32.75 ng/ml respectively.IDO induced by IFN-γ could degrade the intracellular tryptophan to restrict T.gondii growth.
      Conclusion   Determination of IDO activity by using reversed-phase HPLC was a sensitive, simple and reliable method.IDO could restrict T.gondii growth.

     

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