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
em Zh
| WPRIM
| ID: wpr-251800
Biblioteca responsável:
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.
Texto completo:
1
Base de dados:
WPRIM
Assunto principal:
Farmacologia
/
Plasmídeos
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RNA Mensageiro
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Transfecção
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Regiões Promotoras Genéticas
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Genes Reporter
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Colágeno Tipo I
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Avaliação Pré-Clínica de Medicamentos
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Tratamento Farmacológico
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Fator de Crescimento Transformador beta1
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
/
Screening_studies
Limite:
Humans
Idioma:
Zh
Revista:
Acta Pharmaceutica Sinica
Ano de publicação:
2015
Tipo de documento:
Article