Screening of the anti-atherogenic drugs by using THP-1-HIF-1α-HER-Luciferase cell model
10.3969/j.issn.1674-8115.2019.01.007
- Author:
Yu-Jun QIAN
1
Author Information
1. Pharmacy Department, Shanghai Punan Hospital of Pudong New District
- Publication Type:Journal Article
- Keywords:
Atherosclerosis (AS);
Foam cell;
Hypoxia-inducible factor 1α (HIF-1α);
THP-1 cell;
U937 cell
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2019;39(1):33-38
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To screen the anti-atherosclerosis (AS) activity of the compounds by using THP-1-HIF-1α-HER-Luciferase high-throughput model, and to verify the anti-AS function of the effective compounds. Methods: THP-1-HIF-1α-HER-Luciferase cells were pretreated with different concentrations of compounds (1, 10, and 100 μg/mL) for 2 h, then cultured under hypoxia for 24 h. Luciferase activity of cells was detected and compounds with anti-AS activity were screened by luciferase activity evaluation. THP-1 and U937 cells were pretreated with effective compounds, and then induced for 24 h by oxidized low density lipoprotein (OX-LDL). The formation of foam cells was observed by oil red staining. The mRNA level of hypoxia-inducible factor 1α (HIF-1α) was detected by real-time quantitative PCR (qPCR). HIF-1α protein expression was detected by Western blotting. Anti-AS activity of effective compounds were evaluated. Results: Among the 200 compounds, 11 compounds could significantly inhibit the increase of luciferase activity in THP-1-HIF-1α-HER-Luciferase cells induced by hypoxia (all P<0.05), and compound numbered 14 (C14) had the most significant inhibitory effect. THP-1 and U937 cells formed foam cells induced for 24 h by OX-LDL. However, cells were pretreated with C14 for 2 h, which could significantly inhibit the formation of foam cells induced by OX-LDL. Cells were induced for 24 h by OX-LDL, which could significantly increase the expression of HIF-1α mRNA and protein (all P<0.05), while cells pretreated with C14 could significantly inhibit the increase of HIF-1α mRNA and protein expression in a gradient-dependent manner (all P<0.05). Conclusion: THP-1-HIF-1α-HER-Luciferase high-throughput model can be reliability used in screening of compounds with anti-AS activity. C14 has the good anti-AS activity characteristics.
