Background: Transforming growth factor-β (TGF-β) is a major regulator of extracellular matrix (ECM) events, particularly collagen production.
Aim: We explored whether the expression of matrix metalloproteinases (MMPs) and collagen are transcriptionally regulated by the TGF-β and Smad signaling pathways, and the roles played by NF-κB and mitogen-activated protein kinase (MAPK) signaling in normal, aged, human dermal fibroblasts.
Methods: We quantified mRNA and protein expression using real-time PCR and immunoblotting of proteins from cells in passage 5-15.
Results: The levels of mRNAs encoding TGF-β1, TGF-β3, and TGF-β receptor type I (TGFβ RI) decreased with increasing passage number. The levels of mRNAs encoding TGF-β2, TGFβ RII, and TGFβ RIII increased to passage 10 but decreased by passage 15. The levels of mRNAs encoding Smad-2, -3, -4, and -7 decreased with increasing passage number. The level of mRNA encoding MMP-1 increased with increasing passage number, and the levels of mRNAs encoding MMP-2, TIMP-1, and TIMP-2 increased to passage 10 but decreased by passage 15. The levels of mRNAs encoding collagen types I and II decreased with increasing passage number. At the protein level, NF-κB, IκBα, p38, ERK, Akt, and JNK became increasingly phosphorylated at higher passage numbers.
Conclusion: Our results suggest that reductions in the expression levels of MMPs and collagen types I and III in aging human dermal fibroblasts reflect reduced expression of TGF-β/Smad and TGF-β receptors, thus compromising the TGF-β receptor-binding capacity of fibroblasts; the NF-κB and Akt-JNK/MAPK signaling pathways may play active roles in this process.
Keywords: Smad; TGF-β; collagen; matrix metalloproteinases; tissue inhibitor of metalloproteinase.
© 2019 Wiley Periodicals, Inc.