The optimization and application of cell-based screening model for IL-17-mediated signaling pathway
10.16438/j.0513-4870.2019-0703
- VernacularTitle:IL-17信号通路抑制剂细胞筛选模型的优化及应用
- Author:
Ni-na XUE
1
;
Ming JI
1
;
Ming-yi ZHANG
1
;
Yi-chen LIU
1
;
Xiao-guang CHEN
1
Author Information
1. State Key Laboratory of Bioactive Substances and Functions of Natural Medicines/Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
- Publication Type:Research Article
- Keywords:
interleukin-17;
screening model;
autoimmune disease;
tumor immunity
- From:
Acta Pharmaceutica Sinica
2019;54(10):1858-1862
- CountryChina
- Language:Chinese
-
Abstract:
We explore and verify the optimized condition for HEK-Blue IL-17 screening model, and screen the compounds that inhibits IL-17-medited signaling pathway. HEK-Blue IL-17 cells (5×104 cells per well) were seeded into the 96 plates followed by different concentrations of IL-17A or IL-17F alone, or in combination with tested compounds for 16 h. Then, the supernatant medium was incubated with QUANTI-Blue for 1 or 3 h to detect the OD value at λ655nm. The secreted alkaline phosphatase (SEAP) production was an index of IL-17-mediated signaling activation in HEK-Blue IL-17 cells. We found that both IL-17A and IL-17F can significantly activate the IL-17 signaling pathway in HEK-Blue IL-17 cells. The available dosage of IL-17A and IL-17F were 10 and 100 ng·mL-1, respectively. The reaction time of SEAP and QUANTI-Blue was 1 h. In this model, arctigenin and epigallocatechin gallate (EGCG) could inhibit the IL-17A and IL-17F-mediated signaling pathway. This established and optimized screening model of HEK-Blue IL-17 cells was suitable for screening inhibitors of IL-17-mediated signaling pathway.
