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Chinese Journal of Burns ; (6): 284-287, 2007.
Article in Chinese | WPRIM | ID: wpr-347686

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of epidermal growth factor (EGF) on apoptosis induced by TNF-alpha and the expression of PPARbeta in HaCaT keratinocytes.</p><p><b>METHODS</b>HaCaT keratinocytes were cultured and randomly divided into A (normal control), B (with treatment of 10 ng/ml TNF-alpha for 24 hours), C (with treatment of 20 ng/ml TNF-alpha for 24 hours), D (with treatment of 10 ng/ml TNF-alpha after 20 ng/ml EGF treatment for 4 hours), E (with treatment of 20 ng/ml TNF-alpha after 20 ng/ml EGF treatment for 4 hours) groups. The apoptosis of HaCaT keratinocytes was observed by flow cytometry. The proliferative activity of HaCaT keratinocytes was evaluated by MTT method. The activity of caspase-3 was analyzed with caspase colorimetric assay Kit. The changes in the mRNA and protein expression of PPARbeta in HaCaT keratinocytes were observed by RT-PCR and western-blotting after treatment with different concentrations (5, 10, 20, 40 ng/ml) of EGF for 4 or 24 hrs.</p><p><b>RESULTS</b>Compared with A and B groups [(32 +/- 6)%, (57 +/- 6)%], the apoptosis of HaCaT keratinocytes in D and E groups were significantly increased [(20 +/- 3)%, (28 +/- 4)%, respectively, P < 0.01], while the survival rate of HaCaT keratinocytes in D and E groups increased, and the caspase-3 activity were decreased (P < 0.01). The expression of PPARbeta mRNA and protein in HaCaT keratinocytes reached the peak with the treatment of 20 ng/ml EGF.</p><p><b>CONCLUSION</b>EGF can inhibit the apoptosis of HaCaT keratinocytes induced by TNF-alpha, and it can also increase the expression of PPARbeta.</p>


Subject(s)
Humans , Apoptosis , Caspase 3 , Metabolism , Cell Line , Epidermal Growth Factor , Pharmacology , Keratinocytes , Cell Biology , Metabolism , PPAR-beta , Metabolism , RNA, Messenger , Metabolism , Tumor Necrosis Factor-alpha , Pharmacology
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