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KMID : 0356720010170030113
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Journal of the Korean Society of Coloproctology
2001 Volume.17 No. 3 p.113 ~ p.118
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Change of α-SM Actin Expression Induced by the Antibody for TGF-β in Fibroblast NIH3T3 Culture -The basic research for the inhibition of wound contracture-
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Lee Bong-Wha
Kang Hyung-Kil
Kim Jeong-Jin
Kim Dong-Kun
Yoon Dae-Won
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Abstract
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Purpose: The purpose of this experiment is to measure the expression of TGF-beta and alpha-SM actin (smooth muscle actin) from fibroblast culture by the duration of culture days and to analyze the inhibition of alpha-SM actin expression in fibroblast by the antibodies for TGF-beta.
Methods: The levels of alpha-SM actin from the paired NIH3T3 cell cultures with TGF-beta 1 containing medium (10 ng/ml) and with the antibody (for TGF-beta) medium (1 or 2 ug/ml) were determined by SDS PAGE for cell lysate protein, Western blot with ECL autoradiography and immuno - slot blot.
Results: In NIH3T3 culture, the expression of alpha-SM actin increased at culture days 4, 5, 6. TGF-beta was expressed from 2nd day of culture and increased by day 7. The addition of TGF beta (10 ng/ml) did not increased the expression of alpha-SM actin. But alpha-SM actin expression decreased in the presence of anti-TGF beta antibody. The decrease of expression was proportional to the concentration of antibody and duration of exposure to the antibody.
Conclusion: Endogenous TGF-beta produced by fibroblast cultures is sufficient to express the alpha-SM actin from the myofiboblast. There was no additive expression of alpha-SM actin with exogenous TGF-beta 1. The antibody for TGF- beta inhibits the production of the alpha-SM actin during wound healing and may prevent the wound contracture.
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KEYWORD
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Wound healing, Wound contracture, Myofibroblast, TGF-beta, Smooth muscle actin
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