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Functional Implications of the IL-6 Signaling Pathway in Keloid Pathogenesis  Mohammad Ghazizadeh, Mamiko Tosa, Hajime Shimizu, Hiko Hyakusoku, Oichi Kawanami 

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Presentation on theme: "Functional Implications of the IL-6 Signaling Pathway in Keloid Pathogenesis  Mohammad Ghazizadeh, Mamiko Tosa, Hajime Shimizu, Hiko Hyakusoku, Oichi Kawanami "— Presentation transcript:

1 Functional Implications of the IL-6 Signaling Pathway in Keloid Pathogenesis 
Mohammad Ghazizadeh, Mamiko Tosa, Hajime Shimizu, Hiko Hyakusoku, Oichi Kawanami  Journal of Investigative Dermatology  Volume 127, Issue 1, Pages (January 2007) DOI: /sj.jid Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Growth curves of cultured KFs and NFs demonstrated a marked increase in KFs from day 4 of culture (P<0.05). The KFs growth rate was declined to near that of NFs level after inhibition of IL-6Rα by its corresponding antibody. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 (a) RT-PCR semiquantitative mRNA measurements of IL-6, IL-6Rα, and gp130 in the IL-6 signal pathway in four cases each of cultured NFs and KFs show increased levels in KFs compared to NFs. Glyceraldehyde 3-phosphate dehydrogenase serves as an internal control. Data represent mean±SEM of the four cases in each group. (b) Quantification of IL-6 secretion by ELISA shows significant increases in cultured KFs compared to NFs. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Effect of IL-6 peptide or inhibition of IL-6 or IL-6Rα. ECM gene expression by (a) addition of IL-6 peptide or (b) inhibition of IL-6 with anti-IL-6 antibody (c) or IL-6Rα with anti-IL6Rα antibody in serum-free NF or KF cultures show significant dose-dependent increases or decreases in COL1A2 and FN1 mRNAs, respectively (P<0.05). Data represent mean±SEM of triplicate measurements. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Effect of IL-6 induction or inhibition on collagen synthesis. Determination of collagen synthesis by measuring PICPs after induction of IL-6 by IL-1β peptide, and stimulation by IL-6 peptide in cultured NFs, or inhibition of IL-6 or IL-6Rα in cultured KFs demonstrate a dose-dependent increase or decease in PICP respectively (P<0.05 for doses >20 vs 0). Data represent mean±SEM of triplicate measurements in each group. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 mRNA and protein expressions of the downstream targets. RT-PCR semiquantitative mRNA measurements of several downstream targets (JAK1, 286bp; STAT3, 378bp; RAF1, 667bp, and ELK1, 494bp) in the IL-6 signal pathway in four cases each of cultured NFs and KFs show increased levels in KFs compared to NFs. Glyceraldehyde 3-phosphate dehydrogenase (226bp) serves as an internal control. Data represent mean±SEM of the four cases in each group. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 Representative Western blot analysis of three randomly selected cases of cultured KFs and NFs. Equal amounts of total cell lysates from KF and NF cultured cells were size fractionated on SDS-polyacrylamide gels, blotted onto polyvinylidene difluoride membranes and Western blot was performed using antibodies to total or phosphorylated forms for each element. Increased protein expression is observed for phosphorylated/activated and total forms of JAK1, STAT3, RAF1, and ELK1 in KFs compared to NFs. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 Immunoperoxidase stainings of keloid tissues. Hematoxylin and eosin-stained section shows fibroblast proliferation and abundant broad bundles of collagen deposits. KFs frequently showed moderate to strong expressions for IL-6, IL-6Rα, gp130, JAK1, STAT3, RAF1, and ELK1. Inset represents immunostaining in NFs from the section. Bar=200μm. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

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