Induction of Phase II Enzymes Glutathione-S-Transferase and NADPH: Quinone Oxydoreductase 1 with Novel Sulforaphane Derivatives in Human Keratinocytes: Evaluation of the Intracellular GSH Level

Sohely Sikdar^*, Benjamin Lallemand, Jacques Dubois

Laboratoire de Chimie BioAnalytique, Toxicologie et Chimie Physique Appliquée, Faculté de Pharmacie, Université Libre de Bruxelles (ULB), Boulevard du Triomphe, Brussels, Belgium.

DOI: 10.4236/pp.2014.510105                

Abstract

Phase II enzymes including NADPH: Quinone Oxydoreductase 1 (NQO1) and Glutathione-S-Transferase (GST) represents a major and natural cellular protection system against deleterious environmental factors which cause skin damages. Sulforaphane is one of the most popular isothiocyanates found in cruciferous vegetables and known for its cytoprotective effects by inducing Phase II enzymes. Five novel sulforaphane derivatives were synthetized and tested for their activity on NQO1 and GST induction as well as for their effect on total GSH intracellular level using colorimetric assays on human keratinocytes cell line (HaCat). As sulforaphane and the synthetized components showed variable toxicity after their evaluation by means of in vitro cytotoxicity (MTT test), cells were treated at a concentration of 5 μM during 48 hours. The results showed that the addition products of sulforaphane decreased cytotoxity but none of those derivatives had a better effect than referenced sulforaphane on Phase II enzymes. It seems that the isothiacyanate function remains important for the sulforaphane activity.

Keywords

Phase 2, Glutathione, Sulforaphane, Keratinocytes

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Kwak, M.-K., Egner, P.A., Dolan, P.M., Ramos-Gomez, M., Groopman, J.D., Itoh, K., Yamamoto, M. and Kensler, T.W. (2001) Role of Phase 2 Enzyme Induction in Chemoprotection by Dithioethiones. Mutation Research, 480-481, 305-315. http://dx.doi.org/10.1016/S0027-5107(01)00190-7
[2]	Iyanagi, T. (2007) Molecular Mechanism of Phase I and Phase II Drug-Metabolizing Enzymes: Implications for Detoxification. International Review of Cytology, 260, 35-112. http://dx.doi.org/10.1016/S0074-7696(06)60002-8
[3]	Nolan, K.A. Timson, D.J., Stratford, I.J. and Bryce, R.A. (2006) In Silico Identification and Biochemical Characterization of Novel Inhibitors of NQO1. Bioorganic & Medicinal Chemistry Letters, 16, 6246-6254. http://dx.doi.org/10.1016/j.bmcl.2006.09.015
[4]	Dinkova-Kostova, A.T. and Talalay, P. (2010) NAD(P)H:Quinine Acceptor Oxidoreductase 1 (NQO1), a Multifunctional Antioxidant Enzyme and Exceptionally Versatile Cytoprotector. Archives of Biochemistry and Biophysics, 501, 116-123. http://dx.doi.org/10.1016/j.abb.2010.03.019
[5]	Jancova, P., Anzenbacher, P. and Anzenbacherova, E. (2001) Phase II Drug Metabolizing Enzymes. Biomed. Medical Faculty University Palacky Olomouc Czech Republic, 154, 103-116. http://dx.doi.org/10.5507/bp.2010.017
[6]	Lu, S.C. (1999) Regulation of Hepatic Glutathione Synthesis: Current Concepts and Controversies. The FASEB Journal, 13, 1169-1183.
[7]	Suh, J.H., Shenvi, S.V., Dixon, B.M., Liu, H., Jaiswal, A.K., Liu, R.M. and Hagen, T.M. (2004) Decline in Transcriptional Activity of Nrf2 Causes Age-Related Loss of Glutathione Synthesis, Which Is Reversible with Lipoic Acid. Proceedings of the National Academy of Sciences of the United States of America, 101, 3381-3386. http://dx.doi.org/10.1073/pnas.0400282101
[8]	Shenvi, S.V., Smith, E. and Hagen, T.M. (2012) Identification of Age-Specific Nrf2 Binding to a Novel Antioxidant Response Element Locus in the Gclc Promoter: A Compensatory Means for the Loss of Glutathione Synthetic Capacity in the Aging Rat Liver. Aging Cell, 11, 297-304. http://dx.doi.org/10.1111/j.1474-9726.2011.00788.x
[9]	Dinkova-Kostova, A.T., Holtzclaw, W.D. and Kensler, T.W. (2005) The Role of Keap1 in Cellular Protective Responses. Chemical Research in Toxicology, 18, 1779-1791. http://dx.doi.org/10.1021/tx050217c
[10]	Wakabayashi, N., Dinkova-Kostova, A.T., Holtzclaw, W.D., Kang, M.I., Kobayashi, A., Yamamoto, M., Kensler, T. W. and Talalay, P. (2004) Protection against Electrophile and Oxidant Stress by Induction of the Phase 2 Response: Fate of Cysteine of the Keap 1 Sensor Modified by Inducers. Proceedings of the National Academy of Sciences of the United States of America, 101, 2040-2045. http://dx.doi.org/10.1073/pnas.0307301101
[11]	Owen, J.B. and Butterfield, D.A. (2010) Measurement of Oxidized/Reduced Glutathione Ratio. Methods in Molecular Biology, 648, 269-277. http://dx.doi.org/10.1007/978-1-60761-756-3_18
[12]	Shapiro, T.A., Fahey, J.W., Wade, K.L., Stephenson, K.K. and Talalay, P. (2001) Chemoprotective Glucosinolates and Isothiocyanates of Broccoli Sprouts: Metabolism and Excretion in Humans. Cancer Epidemiology, Biomarkers & Prevention, 10, 501-508.
[13]	Emmert, S.W., Desai, D., Amin, S. and Richie Jr., J.P. (2010) Enhanced Nrf2-Dependant Induction of Glutathione in Mouse Embryonic Fibroblasts by Isoselenocyanate Analog of Sulforaphane. Bioorganic & Medicinal Chemistry Letters, 20, 2675-2679. http://dx.doi.org/10.1016/j.bmcl.2010.01.044
[14]	Fahey, J.W. and Talalay, P. (1999) Antioxidant Functions of Sulforaphane: A Potent Inducer of Phase II Detoxication Enzymes. Food and Chemical Toxicology, 37, 973-979. http://dx.doi.org/10.1016/S0278-6915(99)00082-4
[15]	Dinkova-Kostova, A.T., Fahey, J.W., Wade, K.L., Jenkins, S.N., Shapiro, T.A., Fuchs, E.J., Kerns, M.L. and Talalay, P. (2007) Induction of the Phase 2 Response in Mouse and Human Skin by Sulforaphane-Containing Broccoli Sprout Extracts. Cancer Epidemiology, Biomarkers & Prevention, 16, 847-851. http://dx.doi.org/10.1158/1055-9965.EPI-06-0934
[16]	Abel, E.L., Boulware, S., Fields, T., Mclvor, E., Powell, K.L., DiGiovanni, J., Vasquez, K.M. and MacLeod, M.C. (2013) Sulforaphane Induces Phase II Detoxication Enzymes in Mouse Skin and Prevents Mutagenesis Induced by a Mustard Gas Analog. Toxicology and Applied Pharmacology, 266, 439-442. http://dx.doi.org/10.1016/j.taap.2012.11.020
[17]	Lallemand, B., Chaix, F., Bury, M., Bruyère, C., Ghostin, J., Becker, J.P., Delporte, C., Gelbcke, M., Mathieu, V., Dubois, J., Prévost, M., Jabin, I. and Kiss, R. (2011) N-(2-(3[3,5-bis(trifluoromethyl)phényl]ureido)ethyl)-Glycyrrhetinamide (6b): A Novel Anticancer Glycyrrhetinic Acid Derivative that Targets the Proteasome and Displays Anti-Kinase Activity. Journal of Medicinal Chemistry, 54, 6501-6513. http://dx.doi.org/10.1021/jm200285z
[18]	Prochaska, H.J. and Santamaria, A.B. (1988) Direct Measurement of NAD(P)H: Quinone Reductase from Cells Cultured in Microtiter Wells: A Screening Assay for Anticarcinogenic Enzyme Inducers. Analytical Biochemistry, 169, 328-336.
[19]	Ahn, Y.H., Hwang, Y., Liu, H., Wang, X.J., Zhang, Y., Stephenson, K.K., Boronina, T.N., Cole, R.N., Dinkova-Kostova, A.T., Talalay, P. and Cole, P.A. (2010) Electrophilic Tuning of the Chemoprotective Natural Product Sulforaphane. Proceedings of the National Academy of Sciences of the United States of America, 107, 9590-9595. http://dx.doi.org/10.1073/pnas.1004104107