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Int. J. Mol. Sci. 2016, 17(5), 667; doi:10.3390/ijms17050667

The Establishment of an Assay to Measure DNA Polymerase-Catalyzed Repair of UVB-Induced DNA Damage in Skin Cells and Screening of DNA Polymerase Enhancers from Medicinal Plants

1
Research Center, Maruzen Pharmaceuticals Co., Ltd., Onomichi, Hiroshima 722-0062, Japan
2
Graduate School of Agriculture, Shinshu University, Minamiminowa-mura, Kamiina-gun, Nagano 399-4598, Japan
*
Author to whom correspondence should be addressed.
Academic Editor: Chang Won Choi
Received: 5 February 2016 / Revised: 5 April 2016 / Accepted: 25 April 2016 / Published: 4 May 2016
(This article belongs to the Section Bioactives and Nutraceuticals)
View Full-Text   |   Download PDF [1622 KB, uploaded 4 May 2016]   |  

Abstract

An in vitro assay method was established to measure the activity of cellular DNA polymerases (Pols) in cultured normal human epidermal keratinocytes (NHEKs) by modifying Pol inhibitor activity. Ultraviolet (UV) irradiation enhanced the activity of Pols, especially DNA repair-related Pols, in the cell extracts of NHEKs. The optimal ultraviolet B (UVB) exposure dose and culture time to upregulate Pols activity was 100 mJ/cm2 and 4-h incubation, respectively. We screened eight extracts of medicinal plants for enhancement of UVB-exposed cellular Pols activity using NHEKs, and found that rose myrtle was the strongest Pols enhancer. A Pols’ enhancement compound was purified from an 80% ethanol extract of rose myrtle, and piceatannol was isolated by spectroscopic analysis. Induction of Pol activity involved synergy between UVB irradiation and rose myrtle extract and/or piceatannol. Both the extract and piceatannol reduced UVB-induced cyclobutane pyrimidine dimer production, and prevented UVB-induced cytotoxicity. These results indicate that rose myrtle extract and piceatannol, its component, are potential photo-protective candidates for UV-induced skin damage. View Full-Text
Keywords: DNA polymerases; enzyme enhancer; normal human epidermal keratinocytes (NHEK); ultraviolet B (UVB); rose myrtle; piceatannol DNA polymerases; enzyme enhancer; normal human epidermal keratinocytes (NHEK); ultraviolet B (UVB); rose myrtle; piceatannol
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Ikeoka, S.; Nakahara, T.; Iwahashi, H.; Mizushina, Y. The Establishment of an Assay to Measure DNA Polymerase-Catalyzed Repair of UVB-Induced DNA Damage in Skin Cells and Screening of DNA Polymerase Enhancers from Medicinal Plants. Int. J. Mol. Sci. 2016, 17, 667.

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