title: Substance P increases CCN2 dependent on TGF-beta yet Collagen Type I via TGF-beta1 dependent and independent pathways in tenocytes.
abstract: Transforming growth factor beta 1 (TGFbeta-1) and connective tissue growth factor (CCN2) are important mediators of tissue repair and fibrosis, with CCN2 functioning as a downstream mediator of TGFβ-1. Substance P (SP) is also linked to collagen production in tenocytes. A link between SP, TGFbeta-1 and CCN2 has yet to be established in tenocytes or fibrogenic processes. We sought to determine whether SP induces tenocyte proliferation, CCN2, or collagen production via TGFbeta-1 signaling or independently in rat primary tenocytes. Tenocytes were isolated from rat tendons, cultured and stimulated by SP and/or TGFbeta-1. Cultured cells expressed proteins characteristic of tenocytes (vimentin and tenomodulin) and underwent increased proliferation dose dependently after SP and TGFbeta-1 treatments, alone or combined (more than SP alone when combined). SP induced TGFbeta-1 expression in tenocytes in both dose- and time-dependent manners. SP and TGFbeta-1, alone or combined, stimulated CCN2 expression in tenocytes and their supernatants after both 24 and 48 h of stimulation; a response blocked with addition of a TGFbeta-1 receptor inhibitor. In contrast, SP potentiated collagen type I secretion by tenocytes, a response abrogated by the TGFbeta-1 receptor inhibitor after 48 h of stimulation, but not after the shorter 24 h of stimulation. Our findings suggest that both SP and TGFbeta-1 can stimulate tenocyte fibrogenic processes, albeit differently. TGFbeta-1 pathway signaling was involved in CCN2 production at all time points examined, while SP induced collagen type I production independently prior to the onset of signaling through the TGFbeta-1 pathway.
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Nagat
authors.initials: N
authors.lastname: Frara
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Fisher
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Joseph T
authors.initials: JT
authors.lastname: Tarr
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Mamta
authors.initials: M
authors.lastname: Amin
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Steven N
authors.initials: SN
authors.lastname: Popoff
authors.affiliation: a Department of Anatomy and Cell Biology, Lewis Katz School of Medicine , Temple University , Philadelphia , PA , USA.
authors.forename: Mary F
authors.identifier: 0000-0002-5235-9803
authors.initials: MF
authors.lastname: Barbe
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis*; D055513:Connective Tissue Growth Factor / Q000378:metabolism; D005260:Female; D051381:Rats; D017207:Rats, Sprague-Dawley; D000077293:Receptor, Transforming Growth Factor-beta Type I / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013373:Substance P / Q000494:pharmacology*; D000070916:Tenocytes / Q000166:cytology / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
chemical_list: C526316:CCN2 protein, rat; D024042:Collagen Type I; C508962:Tgfb1 protein, rat; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor; D013373:Substance P; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625843:Tgfbr1 protein, rat
keywords: CTGF; Collagen; proliferation; substance P; tendons