FNS  Vol.6 No.17 , December 2015
Low-Dose of the Sulforaphane Precursor Glucoraphanin as a Dietary Supplement Induces Chemoprotective Enzymes in Humans
Abstract: Broccoli sprout (BS) supplements have been marketed for over a decade for the promising health beneficial effects of sulforaphane (SFN), which induces Nrf2 signaling and downstream chemoprotective genes, including phase 2 enzymes. Most commercially available BS supplements encapsulate heat-processed BS containing glucoraphanin (GR), which is hydrolyzed to SFN by the intestinal microbiota. However, the absorption behavior of SFN following the intake of such BS supplements is still unclear. Additionally, the GR dose (around 30 mg) recommended by many manufacturers of BS supplements is relatively lower than the effective dose determined in previous intervention studies. The aims of this study were to assess the effects of a single administration of a typical BS supplement containing lower doses of GR (30 or 60 mg from 3 or 6 capsules, respectively) on SFN absorption, and also to assess the serum activities of phase 2 enzymes as possible surrogate markers of the beneficial effects of SFN. Urinary excreted isothiocyanates and dithiocarbamates showed that the SFN absorption following administration of BS supplement was prolonged and varied among individuals, which conforms to the well-known characteristics of intestinal microbiota-mediated SFN absorption. The amount of SFN absorbed increased dose-dependently but not linear fashion (9.27 μmol and 13.5 μmol for 3 and 6 capsules, respectively). There was no significant difference in SFN bioavailability and the number of capsules consumed. Serum activities of phase 2 enzymes glutathione S-transferase (GST) and NAD(P)H: quinone oxidoreductase 1 (NQO1), which have been reported to display “chemoprotected states” in organs such as the liver, were dose-dependently and synchronously elevated (p < 0.05) following BS supplement intake. This suggests that a low dose of GR (30 mg) exerts chemoprotective effects in humans. In conclusion, our findings will be useful in future clinical studies investigating the chemoprotective effects of SFN, and for the development of BS supplement products.
Cite this paper: Ushida, Y. , Suganuma, H. and Yanaka, A. (2015) Low-Dose of the Sulforaphane Precursor Glucoraphanin as a Dietary Supplement Induces Chemoprotective Enzymes in Humans. Food and Nutrition Sciences, 6, 1603-1612. doi: 10.4236/fns.2015.617165.

[1]   Higdon, J.V., Delage, B., Williams, D.E. and Dashwood, R.H. (2007) Cruciferous Vegetables and Human Cancer Risk: Epidemiologic Evidence and Mechanistic Basis. Pharmacology Research, 55, 224-236.

[2]   Talalay, P. and Fahey, J.W. (2001) Phytochemicals from Cruciferous Plants Protect against Cancer by Modulating Carcinogen Metabolism. Journal of Nutrition, 131, 3027S-3033S.

[3]   Zhao, J. and Zhao, L. (2014) Cruciferous Vegetables Intake Is Associated with Lower Risk of Renal Cell Carcinoma: Evidence from a Meta-Analysis of Observational Studies. PLoS One, 8, e75732.

[4]   Dinkova-Kostova, A.T. (2013) Chemoprotection against Cancer by Isothiocyanates: A Focus on the Animal Models and the Protective Mechanisms. Topics in Current Chemistry, 329, 179-201.

[5]   Zhang, Y., Talalay, P., Cho, C.G. and Posner, G.H. (1992) A Major Inducer of Anticarcinogenic Protective Enzymes from Broccoli: Isolation and Elucidation of Structure. Proceedings of the National Academy of Sciences of the United States of America, 89, 2399-2403.

[6]   McMahon, M., Itoh, K., Yamamoto, M., Chanas, S.A., Henderson, C.J., McLellan, L.I., Wolf, C.R., Cavin, C. and Hayes, J.D. (2001) The Cap'n'Collar Basic Leucine Zipper Transcription Factor Nrf2 (NF-E2 p45-Related Factor 2) Controls Both Constitutive and Inducible Expression of Intestinal Detoxification and Glutathione Biosynthetic Enzymes. Cancer Research, 61, 3299-3307.

[7]   Kensler, T.W., Egner, P.A., Agyeman, A.S., Visvanathan, K., Groopman, J.D., Chen, J.G., Chen, T.Y., Fahey, J.W. and Talalay, P. (2013) Keap1-nrf2 Signaling: A Target for Cancer Prevention by Sulforaphane. Topics in Current Chemistry, 329, 163-177.

[8]   Kensler, T.W., Chen, J.G., Egner, P.A., Fahey, J.W., Jacobson, L.P., Stephenson, K.K., Ye, L., Coady, J.L., Wang, J.B., Wu, Y., Sun, Y., Zhang, Q.N., Zhang, B.C., Zhu, Y.R., Qian, G.S., Carmella, S.G., Hecht, S.S., Benning, L., Gange, S.J., Groopman, J.D. and Talalay, P. (2005) Effects of Glucosinolate-Rich Broccoli Sprouts on Urinary Levels of Aflatoxin-DNA Adducts and Phenanthrene Tetraols in a Randomized Clinical Trial in He Zuo Township, Qidong, People’s Republic of China. Cancer Epidemiology, Biomarkers & Prevention, 14, 2605-2613.

[9]   Ushida, Y. and Talalay, P. (2013) Sulforaphane Accelerates Acetaldehyde Metabolism by Inducing Aldehyde Dehydrogenases: Relevance to Ethanol Intolerance. Alcohol and Alcoholism, 48, 526-534.

[10]   Toyama, T., Shinkai, Y., Yasutake, A., Uchida, K., Yamamoto, M. and Kumagai, Y. (2011) Isothiocyanates Reduce Mercury Accumulation via an Nrf2-Dependent Mechanism during Exposure of Mice to Methylmercury. Environmental Health Perspectives, 119, 1117-1122.

[11]   Egner, P.A., Chen, J.G., Zarth, A.T., Ng, D.K., Wang, J.B., Kensler, K.H., Jacobson, L.P., Muñoz, A., Johnson, J.L., Groopman, J.D., Fahey, J.W., Talalay, P., Zhu, J., Chen, T.Y., Qian, G.S., Carmella, S.G., Hecht, S.S. and Kensler, T.W. (2014) Rapid and Sustainable Detoxication of Airborne Pollutants by Broccoli Sprout Beverage: Results of a Randomized Clinical Trial in China. Cancer Prevention Research (Philadelphia), 7, 813-823.

[12]   Kensler, T.W., Ng, D., Carmella, S.G., Chen, M., Jacobson, L.P., Muñoz, A., Egner, P.A., Chen, J.G., Qian, G.S., Chen, T.Y., Fahey, J.W., Talalay, P., Groopman, J.D., Yuan J. and Hecht, S.S. (2012) Modulation of the Metabolism of Airborne Pollutants by Glucoraphanin-Rich and Sulforaphane-Rich Broccoli Sprout Beverages in Qidong, China. Carcinogenesis, 33, 101-107.

[13]   Gaona-Gaona, L., Molina-Jijón, E., Tapia, E., Zazueta, C., Hernández-Pando, R., Calderón-Oliver, M., Zarco-Márquez, G., Pinzón, E. and Pedraza-Chaverri, J. (2011) Protective Effect of Sulforaphane Pretreatment against Cisplatin-Induced Liver and Mitochondrial Oxidant Damage in Rats. Toxicology, 286, 20-27.

[14]   Talalay, P., Fahey, J.W., Holtzclaw, W.D., Prestera, T. and Zhang, Y. (1995) Chemoprotection against Cancer by Phase 2 Enzyme Induction. Toxicology Letters, 82-83, 173-179.

[15]   Lenzi, M., Fimognari, C. and Hrelia, P. (2014) Sulforaphane as a Promising Molecule for Fighting Cancer. Cancer Treatment and Research, 159, 207-223.

[16]   Sayed, R.H., Khalil, W.K., Salem, H.A., Kenawy, S.A. and El-Sayeh, B.M. (2014) Sulforaphane Increases the Survival Rate in Rats with Fulminant Hepatic Failure Induced by D-Galactosamine and Lipopolysaccharide. Nutrition Research, 34, 982-989.

[17]   Zhang, Z., Wang, S., Zhou, S., Yan, X., Wang, Y., Chen, J., Mellen, N., Kong, M., Gu, J., Tan, Y., Zheng, Y. and Cai, L. (2014) Sulforaphane Prevents the Development of Cardiomyopathy in Type 2 Diabetic Mice Probably by Reversing Oxidative Stress-Induced Inhibition of LKB1/AMPK Pathway. Journal of Molecular and Cellular Cardiology, 77, 42-52.

[18]   Shirai, Y., Fujita, Y. and Hashimoto, K. (2012) Effects of the Antioxidant Sulforaphane on Hyperlocomotion and Prepulse Inhibition Deficits in Mice after Phencyclidine Administration. Clinical Psychopharmacology and Neuroscience, 10, 94-98.

[19]   Shirai, Y., Fujita, Y., Hashimoto, R., Ohi, K., Yamamori, H., Yasuda, Y., Ishima, T., Suganuma, H., Ushida, Y., Takeda, M. and Hashimoto, K. (2015) Dietary Intake of Sulforaphane-Rich Broccoli Sprout Extracts during Juvenile and Adolescence Can Prevent Phencyclidine-Induced Cognitive Deficits at Adulthood. PLoS One, 10, e0127244.

[20]   Gao, X. and Talalay, P. (2004) Induction of Phase 2 Genes by Sulforaphane Protects Retinal Pigment Epithelial Cells against Photooxidative Damage. Proceeding of National Academy Sciences of the United States of America, 101, 10446-10451.

[21]   Talalay, P., Fahey, J.W., Healy, Z.R., Wehage, S.L., Benedict, A.L., Min, C. and Dinkova-Kostova, A.T. (2007) Sulforaphane Mobilizes Cellular Defenses That Protect Skin against Damage by UV Radiation. Proceeding of National Academy Sciences of the United States of America, 104, 17500-17505.

[22]   Singh, K., Connors, S.L., Macklin, E.A., Smith, K.D., Fahey, J.W., Talalay, P. and Zimmerman, A.W. (2014) Sulforaphane Treatment of Autism Spectrum Disorder (ASD). Proceeding of National Academy Sciences of the United States of America, 111, 15550-15555.

[23]   Bahadoran, Z., Mirmiran, P., Hosseinpanah, F., Rajab, A., Asghari, G. and Azizi, F. (2012) Broccoli Sprouts Powder Could Improve Serum Triglyceride and Oxidized LDL/LDL-Cholesterol Ratio in Type 2 Diabetic Patients: A Randomized Double-Blind Placebo-Controlled Clinical Trial. Diabetes Research and Clinical Practice, 96, 348-354.

[24]   Yanaka, A., Fahey, J.W., Fukumoto, A., Nakayama, M., Inoue, S., Zhang, S., Tauchi, M., Suzuki, H., Hyodo, I. and Yamamoto, M. (2009) Dietary Sulforaphane-Rich Broccoli Sprouts Reduce Colonization and Attenuate Gastritis in Helicobacter Pylori-Infected Mice and Humans. Cancer Prevention Research (Philadelphia), 2, 353-360.

[25]   Kikuchi, M., Ushida, Y., Shiozawa, H., Umeda, R., Tsuruya, K., Aoki, Y., Suganuma, H. and Nishizaki, Y. (2015) Sulforaphane-Rich Broccoli Sprout Extract Improves Hepatic Abnormalities in Male Subjects. World Journal of Gastroenterology, in Press.

[26]   Fahey, J.W., Zhang, Y. and Talalay, P. (1997) Broccoli Sprouts: An Exceptionally Rich Source of Inducers of Enzymes That Protect against Chemical Carcinogens. Proceeding of National Academy Sciences of the United States of America, 94, 10367-10372.

[27]   Shapiro, T.A., Fahey, J.W., Wade, K.L., Stephenson, K.K. and Talalay, P. (1998) Human Metabolism and Excretion of Cancer Chemoprotective Glucosinolates and Isothiocyanates of Cruciferous Vegetables. Cancer Epidemiology, Biomarkers & Prevention, 7, 1091-1100.

[28]   Cramer, J.M. and Jeffery, E.H. (2011) Sulforaphane Absorption and Excretion Following Ingestion of a Semi-Purified Broccoli Powder Rich in Glucoraphanin and Broccoli Sprouts in Healthy Men. Nutrition and Cancer, 63, 196-201.

[29]   Fahey, J.W., Wehage, S.L., Holtzclaw, W.D., Kensler, T.W., Egner, P.A., Shapiro, T.A. and Talalay, P. (2012) Protection of Humans by Plant Glucosinolates: Efficiency of Conversion of Glucosinolates to Isothiocyanates by the Gastrointestinal Microflora. Cancer Prevention Research (Philadelphia), 5, 603-611.

[30]   Egner, P.A., Chen, J.G., Wang, J.B., Wu, Y., Sun, Y., Lu, J.H., Zhu, J., Zhang, Y.H., Chen, Y.S., Friesen, M.D., Jacobson, L.P., Muñoz, A., Ng, D., Qian, G.S., Zhu, Y.R., Chen, T.Y., Botting, N.P., Zhang, Q., Fahey, J.W., Talalay. P., Groopman. J.D. and Kensler, T.W. (2011) Bioavailability of Sulforaphane from Two Broccoli Sprout Beverages: Results of a Short-Term, Cross-Over Clinical Trial in Qidong, China. Cancer Prevention Research (Philadelphia), 4, 384-395.

[31]   Cramer, J.M., Teran-Garcia, M. and Jeffery, E.H. (2012) Enhancing Sulforaphane Absorption and Excretion in Healthy Men through the Combined Consumption of Fresh Broccoli Sprouts and a Glucoraphanin-Rich Powder. British Journal of Nutrition, 107, 1333-1338.

[32]   Atwell, L.L., Hsu, A., Wong, C.P., Stevens, J.F., Bella, D., Yu, T., Pereira, C.B., Löhr, C.V., Christensen, J.M., Dashwood, R.H., Williams, D.E., Shannon, J. and Ho, E. (2015) Absorption and Chemopreventive Targets of Sulforaphane in Humans Following Consumption of Broccoli Sprouts or a Myrosinase-Treated Broccoli Sprout Extract. Molecular Nutrition & Food Research, 59, 424-433.

[33]   Conaway, C.C., Getahun, S.M., Liebes, L.L., Pusateri, D.J., Topham, D.K., Botero-Omary, M. and Chung, F.L. (2000) Disposition of Glucosinolates and Sulforaphane in Humans after Ingestion of Steamed and Fresh Broccoli. Nutrition and Cancer, 38, 168-178.

[34]   Vermeulen, M., Klöpping-Ketelaars, I.W., van den Berg, R. and Vaes, W.H. (2008) Bioavailability and Kinetics of Sulforaphane in Humans after Consumption of Cooked versus Raw Broccoli. Journal of Agricultural and Food Chemistry, 56, 10505-10509.

[35]   Riedl, M.A., Saxon, A. and Diaz-Sanchez, D. (2009) Oral Sulforaphane Increases Phase II Antioxidant Enzymes in the Human Upper Airway. Clinical Immunology, 130, 244-251.

[36]   Habig, W.H., Pabst, M.J. and Jakoby, W.B. (1974) Glutathione S-Transferases. The First Enzymatic Step in Mercapturic Acid Formation. Journal of Biological Chemistry, 249, 7130-7139.

[37]   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.

[38]   Fahey, J.W., Dinkova-Kostova, A.T., Stephenson, K.K. and Talalay, P. (2004) The “Prochaska” Microtiter Plate Bioassay for Inducers of NQO1. Methods in Enzymology, 382, 243-258.

[39]   Agudo, A., Ibáñez, R., Amiano, P., Ardanaz, E., Barricarte, A., Berenguer, A., Dolores Chirlaque, M., Dorronsoro, M., Jakszyn, P., Larrañaga, N., Martinez, C., Navarro, C., Pera, G., Quirós, J.R., Sanchéz, M.J., Tormo, M.J. and González, C.A. (2008) Consumption of Cruciferous Vegetables and Glucosinolates in a Spanish Adult Population. European Journal of Clinical Nutrition, 62, 324-331.

[40]   Sones, K., Heaney, R.K. and Fenwick, G.R. (1984) An Estimate of the Mean Daily Intake of Glucosinolates from Cruciferous Vegetables in the UK. Journal of the Science of Food and Agriculture, 35, 712-720.

[41]   Prochaska, H.J. and Fernandes, C.L. (1993) Elevation of Serum Phase II Enzymes by Anticarcinogenic Enzyme Inducers: Markers for a Chemoprotected State? Carcinogenesis, 14, 2441-2445.

[42]   Nijhoff, W.A., Mulder, T.P., Verhagen, H., van Poppel, G. and Peters, W.H. (1995) Effects of Consumption of Brussels Sprouts on Plasma and Urinary Glutathione S-Transferase Class-Alpha and -Pi in Humans. Carcinogenesis, 16, 955-957.

[43]   Bogaards, J.J., Verhagen, H., Willems, M.I., van Poppel, G. and van Bladeren, P.J. (1994) Consumption of Brussels Sprouts Results in Elevated Alpha-Class Glutathione S-Transferase Levels in Human Blood Plasma. Carcinogenesis, 15, 1073-1075.

[44]   Chow, H.H., Hakim, I.A., Vining, D.R., Crowell, J.A., Tome, M.E., Ranger-Moore, J., Cordova, C.A., Mikhael, D.M., Briehl, M.M. and Alberts, D.S. (2007) Modulation of Human Glutathione S-Transferases by Polyphenon E Intervention. Cancer Epidemiology, Biomarkers & Prevention, 16, 1662-1666.