NADPH oxidase-dependent formation of reactive oxygen species contributes to transforming growth factor β1-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells, and the role of astragalus intervention / 中国结合医学杂志
Chinese journal of integrative medicine
;
(12): 667-674, 2014.
Article
in English
| WPRIM
| ID: wpr-293344
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidasedependent formation of reactive oxygen species (ROS) in the transforming growth factor β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) in rat peritoneal mesothelial cells (RPMCs), and the effect of Astragalus injection (AGI) intervention.</p><p><b>METHODS</b>Primary RPMCs were cultured to the second generation in vitro. After synchronization for 24 h, the cells were randomly assigned to the following groups control (Group A), AGI (2 g/mL; Group B), TGF-β1 (10 ng/mL; Group C), TGF-β1 (10 ng/mL) + AGI (2 g/mL; Group D; pretreated for 1 h with AGI before TGF-β1 stimulation). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis were employed to evaluate the mRNA and protein expression of the NADPH oxidase subunit p67phox, α-smooth muscle actin (α-SMA) and E-cadherin. The dichlorofluorescein-sensitive cellular ROS levels were measured by a fluorometric assay and confocal microscopy.</p><p><b>RESULTS</b>TGF-β1 significantly induced NADPH oxidase subunit p67phox mRNA and protein expression in RPMCs, as well as inducing the production of intracellular ROS. AGI inhibited this TGF-β1-induced up-regulation by 39.3% and 47.8%, respectively (P<0.05), as well as inhibiting the TGF-β1-induced ROS generation by 56.3% (P<0.05). TGF-β1 also induced α-SMA mRNA and protein expression, and down-regulated E-cadherin mRNA and protein expression (P<0.05). This effect was suppressed by AGI (P<0.05).</p><p><b>CONCLUSIONS</b>NADPH oxidase-dependent formation of ROS may mediate the TGF-β1-dependent EMT in RPMCs. AGI could inhibit this process, providing a theoretical basis for AGI in the prevention of peritoneal fibrosis.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peritoneal Cavity
/
Physiology
/
Base Sequence
/
Polymerase Chain Reaction
/
Reactive Oxygen Species
/
DNA Primers
/
NADPH Oxidases
/
Reverse Transcriptase Polymerase Chain Reaction
/
Cell Biology
/
Epithelium
Limits:
Animals
Language:
English
Journal:
Chinese journal of integrative medicine
Year:
2014
Type:
Article
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