Embryonic stem cell differentiation into smooth muscle cells is mediated by Nox4-produced H2O2

Autor: Zhenling Luo, Lingfang Zeng, Qingbo Xu, Anna Elena Pepe, Andriani Margariti, Qingzhong Xiao
Rok vydání: 2008
Předmět:
Collagen Type IV
Serum Response Factor
Vascular smooth muscle
Time Factors
Physiology
Cellular differentiation
Cell
Myocytes
Smooth Muscle
Receptor
Transforming Growth Factor-beta Type I
Protein Serine-Threonine Kinases
Transfection
Gene Expression Regulation
Enzymologic
Cell Line
Transforming Growth Factor beta1
Mice
medicine
Animals
Phosphorylation
RNA
Small Interfering
Embryonic Stem Cells
NADPH oxidase
biology
urogenital system
NOX4
Gene Expression Regulation
Developmental
NADPH Oxidases
Nuclear Proteins
Cell Differentiation
Cell Biology
Hydrogen Peroxide
musculoskeletal system
Embryonic stem cell
Cell biology
Up-Regulation
Autocrine Communication
Protein Transport
medicine.anatomical_structure
Phenotype
Myocardin
NADPH Oxidase 4
Immunology
cardiovascular system
biology.protein
Trans-Activators
RNA Interference
Stem cell
tissues
Receptors
Transforming Growth Factor beta
Zdroj: American journal of physiology. Cell physiology. 296(4)
ISSN: 0363-6143
Popis: NADPH oxidase (Nox4) produces reactive oxygen species (ROS) that are important for vascular smooth muscle cell (SMC) behavior, but the potential impact of Nox4 in stem cell differentiation is unknown. When mouse embryonic stem (ES) cells were plated on collagen IV-coated dishes/flasks, a panel of SMC-specific genes was significantly and consistently upregulated. Nox4 expression was markedly correlated with such a gene induction as confirmed by real-time PCR, immunofluorescence, and Western blot analysis. Overexpression of Nox4 specifically resulted in increased SMC marker production, whereas knockdown of Nox4 induced a decrease. Furthermore, SMC-specific transcription factors, including serum response factor (SRF) and myocardin were activated by Nox4 gene expression. Moreover, Nox4 was demonstrated to drive SMC differentiation through generation of H2O2. Confocal microscopy analysis indicates that SRF was translocated into the nucleus during SMC differentiation in which SRF was phosphorylated. Additionally, autosecreted transforming growth factor (TGF)-β1 activated Nox4 and promoted SMC differentiation. Interestingly, cell lines generated from stem cells by Nox4 transfection and G418 selection displayed a characteristic of mature SMCs, including expression of SMC markers and cells with contractile function. Thus we demonstrate for the first time that Nox4 is crucial for SMC differentiation from ES cells, and enforced Nox4 expression can maintain differentiation status and functional features of stem cell-derived SMCs, highlighting its impact on vessel formation in vivo and vascular tissue engineering in the future.
Databáze: OpenAIRE
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