Establishment and application of a high-throughput drug screening model based on COL1A1 promoter for anti-liver fibrosis / 药学学报
Acta Pharmaceutica Sinica
; (12): 169-173, 2015.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-251800
Responsible library:
WPRO
ABSTRACT
For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Plasmids
/
RNA, Messenger
/
Transfection
/
Promoter Regions, Genetic
/
Genes, Reporter
/
Collagen Type I
/
Drug Evaluation, Preclinical
/
Drug Therapy
/
Transforming Growth Factor beta1
Type of study:
Diagnostic study
/
Prognostic study
/
Screening study
Limits:
Humans
Language:
Chinese
Journal:
Acta Pharmaceutica Sinica
Year:
2015
Document type:
Article