Young Il Kim1, Kyung Sook Kim2, Hye-Jin Ahn3, In-Hye Kang3, Min Kyung Shin3. 1. Medical Science Research Institute, Kyung Hee University Medical Center, Seoul, Korea. 2. Department of Biomedical Engineering, School of Medicine, Kyung Hee University, Seoul, Korea. 3. Department of Dermatology, School of Medicine, Kyung Hee University, Seoul, Korea.
Abstract
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.
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.