| Change of alpha-SM Actin Expression Induced by the Antibody for TGF-beta in Fibroblast NIH3T3 Culture: The basic research for the inhibition of wound contracture. |
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Lee, Bong Hwa , Kang, Hyung Kil , Kim, Jeong Jin , Kim, Dong Kun , Yoon, Dae Won |
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Department of Surgery, Hallym University College of Medicine, Seoul, Korea. bshlee@hallym.or.kr |
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| Abstract |
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.
CONCLUSIONS
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. |
| Key Words:
Wound healing; Wound contracture; Myofibroblast; TGF-beta; Smooth muscle actin |
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