GCDCA down-regulates gene expression by increasing Sp1 binding to the NOS-3 promoter in an oxidative stress dependent manner

Autor: José Luis Montero-Álvarez, Clara I. Linares, Sandra González-Rubio, Patricia Aguilar-Melero, Antonio Rodríguez-Ariza, Marta Casado, Manuel Rodríguez-Perálvarez, Manuel de la Mata, Ana Julia Fernández-Alvarez, Rafael Sánchez-Sánchez, Gustavo Ferrín, Laura M. López-Sánchez, Juan R. Muñoz-Castañeda, Jordi Muntané
Rok vydání: 2015
Předmět:
Male
Nitric Oxide Synthase Type III
Sp1 Transcription Factor
GLYCOCHENODEOXYCHOLIC ACID
Molecular Sequence Data
Down-Regulation
Oxidative phosphorylation
Biology
medicine.disease_cause
Biochemistry
Ciencias Biológicas
chemistry.chemical_compound
Glycochenodeoxycholic Acid
Cholestasis
Cell Line
Tumor
Gene expression
Glycochenodeoxycholic acid
medicine
Animals
Humans
Rats
Wistar
OXIDATIVE STRESS
Promoter Regions
Genetic
Glyceraldehyde 3-phosphate dehydrogenase
Pharmacology
Sp1 transcription factor
MitoQ
Base Sequence
TRANSCRIPTION FACTOR SP1
CHOLESTATIC LIVER DISEASE
DNA
Bioquímica y Biología Molecular
medicine.disease
Rats
ENDOTHELIAL NITRIC OXIDE SYNTHASE
Cell biology
Oxidative Stress
Gene Expression Regulation
chemistry
biology.protein
CIENCIAS NATURALES Y EXACTAS
Oxidative stress
Protein Binding
Zdroj: Biochemical Pharmacology. 96:39-51
ISSN: 0006-2952
Popis: During the course of cholestatic liver diseases, the toxic effect of bile acids accumulation has been related to the decreased expression of endothelial nitric oxide synthase (NOS-3) and cellular oxidative stress increase. In the present study, we have investigated the relationship between these two biological events. In the human hepatocarcinoma cell line HepG2, cytotoxic response to GCDCA was characterized by the reduced activity of the respiratory complexes II. +. III, the increased expression and activation of the transcription factor Sp1, and a higher binding capacity of this at positions -1386, -632 and -104 of the NOS-3 promoter (pNOS-3). This was associated with a decreased promoter activity and a consequent reduction of NOS-3 expression. The use of antioxidants in GCDCA-treated cells caused a lower activation of Sp1 and the recovery of the pNOS-3 activity and NOS-3 expression and activity. Similarly, the specific inhibition of Sp1 resulted in the improvement of NOS-3 expression. Both, antioxidant treatment and Sp1 inhibition were associated with the reduction of cell death-related parameters. Bile duct ligation in rats confirmed in vitro results concerning the activation of Sp1 and the reduction of NOS-3 expression. Our results provide direct evidence for the involvement of Sp1 in the regulation of NOS-3 expression during cholestasis. Thus, the identification of Sp1 as a potential negative regulator of NOS-3 expression represents a new mechanism by which the accumulation of bile acids causes a cytotoxic effect through the oxidative stress increase, and provides a new potential target in cholestatic liver diseases. Fil: González Rubio, Sandra. Universidad de Córdoba; España Fil: López Sánchez, Laura. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España Fil: Muñoz Castañeda, Juan. Universidad de Córdoba; España Fil: Linares, Clara I.. Universidad de Córdoba; España Fil: Aguilar Melero, Patricia. Universidad de Córdoba; España Fil: Rodríguez Perálvarez, Manuel. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España Fil: Sánchez Sánchez, Rafael. Reina Sofia University Hospital; España Fil: Fernández Alvarez, Ana Julia. Biomedicine Institute of Valencia; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Casado, Marta. Universidad Carlos III de Madrid. Instituto de Salud; España Fil: Montero Álvarez, Jose L.. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España Fil: Rodríguez Ariza, Antonio. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España Fil: Muntané, Jordi. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España Fil: de la Mata, Manuel. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ferrín, Gustavo. Universidad de Córdoba; España. Universidad Carlos III de Madrid. Instituto de Salud; España
Databáze: OpenAIRE
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