TGF-β1 and serum both stimulate contraction but differentially affect apoptosis in 3D collagen gels

Tetsu Kobayashi, Xiang-de Liu, Hui Jung Kim, Tadashi Kohyama, Fu Qiang Wen, Shinji Abe, Qiuhong Fang, Yu Kui Zhu, John R. Spurzem, Peter Bitterman, Stephen I. Rennard

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Apoptosis of fibroblasts may be key for the removal of cells following repair processes. Contraction of three-dimensional collagen gels is a model of wound healing and remodeling. Here two potent inducers of contraction, TGF-β1 and fetal calf serum (FCS) were evaluated for their effect on fibroblast apoptosis in contracting collagen gels. Human fetal lung fibroblasts were cultured in floating type I collagen gels, exposed to TGF-β1 or FCS, and allowed to contract for 5 days. Apoptosis was evaluated using TUNEL and confirmed by DNA content profiling. Both TGF-β1 and serum significantly augmented collagen gel contraction. TGF-β1 also increased apoptosis assessed by TUNEL positivity and DNA content analysis. In contrast, serum did not affect apoptosis. TGF-β1 induction of apoptosis was associated with augmented expression of Bax, a pro-apoptotic member of the Bax/Bcl-2 family, inhibition of Bcl-2, an anti-apoptotic member of the same family, and inhibition of both cIAP-1 and XIAP, two inhibitors of the caspase cascade. Serum was associated with an increase in cIAP-1 and Bcl-2, anti-apoptotic proteins. Interestingly, serum was also associated with an apparent increase in Bax, a pro-apoptotic protein. Blockade of Smad3 with either siRNA or by using murine fibroblasts deficient in Smad3 resulted in a lack of TGF-β induction of augmented contraction and apoptosis. Contraction induced by different factors, therefore, may be differentially associated with apoptosis, which may be related to the persistence or resolution of the fibroblasts that accumulate following injury.

Original languageEnglish (US)
Article number141
JournalRespiratory Research
Volume6
DOIs
StatePublished - Dec 2 2005

Fingerprint

Collagen
Gels
Apoptosis
Serum
Fibroblasts
Apoptosis Regulatory Proteins
In Situ Nick-End Labeling
Caspase Inhibitors
DNA Fingerprinting
Collagen Type I
Wound Healing
Small Interfering RNA
Lung
DNA
Wounds and Injuries

Keywords

  • Apoptosis
  • Fibrosis
  • Gel contraction
  • Transforming growth factor-beta
  • Wound repair

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

TGF-β1 and serum both stimulate contraction but differentially affect apoptosis in 3D collagen gels. / Kobayashi, Tetsu; Liu, Xiang-de; Kim, Hui Jung; Kohyama, Tadashi; Wen, Fu Qiang; Abe, Shinji; Fang, Qiuhong; Zhu, Yu Kui; Spurzem, John R.; Bitterman, Peter; Rennard, Stephen I.

In: Respiratory Research, Vol. 6, 141, 02.12.2005.

Research output: Contribution to journalArticle

Kobayashi, T, Liu, X, Kim, HJ, Kohyama, T, Wen, FQ, Abe, S, Fang, Q, Zhu, YK, Spurzem, JR, Bitterman, P & Rennard, SI 2005, 'TGF-β1 and serum both stimulate contraction but differentially affect apoptosis in 3D collagen gels', Respiratory Research, vol. 6, 141. https://doi.org/10.1186/1465-9921-6-141
Kobayashi, Tetsu ; Liu, Xiang-de ; Kim, Hui Jung ; Kohyama, Tadashi ; Wen, Fu Qiang ; Abe, Shinji ; Fang, Qiuhong ; Zhu, Yu Kui ; Spurzem, John R. ; Bitterman, Peter ; Rennard, Stephen I. / TGF-β1 and serum both stimulate contraction but differentially affect apoptosis in 3D collagen gels. In: Respiratory Research. 2005 ; Vol. 6.
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AU - Wen, Fu Qiang

AU - Abe, Shinji

AU - Fang, Qiuhong

AU - Zhu, Yu Kui

AU - Spurzem, John R.

AU - Bitterman, Peter

AU - Rennard, Stephen I.

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AB - Apoptosis of fibroblasts may be key for the removal of cells following repair processes. Contraction of three-dimensional collagen gels is a model of wound healing and remodeling. Here two potent inducers of contraction, TGF-β1 and fetal calf serum (FCS) were evaluated for their effect on fibroblast apoptosis in contracting collagen gels. Human fetal lung fibroblasts were cultured in floating type I collagen gels, exposed to TGF-β1 or FCS, and allowed to contract for 5 days. Apoptosis was evaluated using TUNEL and confirmed by DNA content profiling. Both TGF-β1 and serum significantly augmented collagen gel contraction. TGF-β1 also increased apoptosis assessed by TUNEL positivity and DNA content analysis. In contrast, serum did not affect apoptosis. TGF-β1 induction of apoptosis was associated with augmented expression of Bax, a pro-apoptotic member of the Bax/Bcl-2 family, inhibition of Bcl-2, an anti-apoptotic member of the same family, and inhibition of both cIAP-1 and XIAP, two inhibitors of the caspase cascade. Serum was associated with an increase in cIAP-1 and Bcl-2, anti-apoptotic proteins. Interestingly, serum was also associated with an apparent increase in Bax, a pro-apoptotic protein. Blockade of Smad3 with either siRNA or by using murine fibroblasts deficient in Smad3 resulted in a lack of TGF-β induction of augmented contraction and apoptosis. Contraction induced by different factors, therefore, may be differentially associated with apoptosis, which may be related to the persistence or resolution of the fibroblasts that accumulate following injury.

KW - Apoptosis

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