Apocynin inhibits the upregulation of TGF-β1 expression and ROS production induced by TGF-β in skeletal muscle cells

Johanna Abrigo, María Gabriela Morales, Felipe Simon, Daniel Cabrera, Gabriella Di Capua, Claudio Cabello-Verrugio*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

16 Scopus citations


Abstract Background Pure apocynin, which can be traditionally isolated and purified from several plant species such as Picrorhiza kurroa Royle ex Benth (Scrophulariaceae), acts as an inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) activity inhibiting its production of reactive oxygen species (ROS). Transforming growth factor type beta 1 (TGF-β1) is a growth factor that produces inhibition of myogenesis, diminution of regeneration and induction of atrophy in skeletal muscle. The typical signalling that is activated by TGF-β involves the Smad pathway. Purpose To evaluate the effect of TGF-β and the effect of apocynin on TGF-β1 expression in skeletal muscle cells. Study design Controlled laboratory study. In vitro assays were performed with C2C12 cells incubated with TGF-β1 in presence or absence of apocynin (NOX inhibitor), SB525334 (TGF-β-receptor I inhibitor), or chelerythrine (PKC inhibitor). Methods TGF-β1 and atrogin-1 expression was evaluated by RT-qPCR and/or ELISA; Smad3 phosphorylation by western blot; Smad4 nuclear translocation by indirect immunofluorescence; and ROS levels by DCF probe fluorescent measurements. Results We show that myoblasts respond to TGF-β1 by increasing its own gene expression in a time- and dose-dependent fashion which was abolished by SB525334 and siRNA for Smad2/3. TGF-β1 also induced ROS. Remarkably, apocynin inhibited the TGF-β1 induced ROS as well as the autoinduction of TGF-β1 gene expression. We also show that TGF-β-induced ROS production and TGF-β1 expression require PKC activity as indicated by the inhibition using chelerythrine. Conclusion These results strongly suggest that TGF-β induces its own expression through a TGF-β-receptor/Smad-dependent mechanism and apocynin is able to inhibit this process, suggesting that requires NOX-induced ROS in skeletal muscle cells.

Original languageEnglish
Article number51860
Pages (from-to)885-893
Number of pages9
Issue number10
StatePublished - 27 Jul 2015
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2015 Elsevier GmbH.


  • Atrophy
  • NOX
  • ROS
  • Skeletal muscle
  • Smad
  • TGF-β1


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