• Embed Doc
  • Copy Link
  • Readcast
  • Collections
  • CommentGo Back
 
Role of Dietary Supplements/Nutraceuticals in ChemopreventionThrough Induction of Cytoprotective Enzymes
Jed W. Fahey
1,2,#and
Thomas W. Kensler
1,*,31
Department of Pharmacology and Molecular Sciences, Johns Hopkins University, Baltimore, MD 21205
2
Department of International Health, Johns Hopkins University, Baltimore, MD 21205
3
Department of Environmental Health Sciences, Johns Hopkins University, Baltimore, MD 21205
Rationale for Enzyme Induction as a Chemopreventive Strategy
Numerous epidemiological studies from many parts of the world report strikingly lower cancerrisks among individuals who consume large quantities of fruit and vegetables (reviewed in(1,2)). As a consequence, a great variety of foods and supplements have been implicated asbeing sources of protective phytochemical factors. These factors can be used as discretechemicals, dietary supplements, or as functional foods. Others have contrasted the fact thatdietary supplements are generally considered to be time-tested, but in large part scientificallyunproven, whereas functional foods are components of the normal diet that are increasinglyshown to have inherent value for maintaining human health (3). The popular literaturerepeatedly highlights some of these phytochemicals more than others (
e.g., capsaicin
frompeppers,
coumarins
from citrus and tomatoes,
epigallocatechin-3-gallate
(
 EGCG
) from greentea,
genistein
from soybeans,
indoles
from broccoli and cabbage,
isothiocyanates
fromcruciferous vegetables,
lycopene
from tomatoes and red grapefruit,
allicin
from garlic andonions,
triterpenoids
from licorice root and citrus,
 pectin
from grapefruit,
resveratrol
fromgrapes,
carotenoids
from red grapefruit,
quercetin
from onions and broccoli, and
 flavonoids
from a variety of fruits and vegetables). The mechanisms responsible for the possible protectiveeffects derived from the consumption of these foods are multiple, probably involve complexinteractions and are incompletely understood. Nonetheless, there is extensive literaturedetailing the actions of specific phytochemicals as well as the intact plants themselves towardsaltering the expression of genes that reflect an adaptive stress response which enhances normalcell survival, and may also promote apoptosis or cell cycle arrest in tumor cells. In the face of electrophilic and oxidative insults, these inducible gene products have been shown to facilitatethe conjugation of xenobiotics, the nucleophilic trapping of activated electrophilic xenobiotics,as well as to increase overall antioxidative capacity in cells, animals and in some cases humans(4–8). Moreover, substantive experimental evidence in animals has been developed to supportthe view that the coordinated induction of these cytoprotective enzymes is a critical andsufficient mechanism to engender protection against the toxic and carcinogenic actions of reactive intermediates (reviewed in (6,9)). As a consequence, monitoring for inducers of thisstress response can be an informative means to identify plants and constituent phytochemicalsof potential chemopreventive utility.
*To whom correspondence should be addressed: Dr. Thomas W. Kensler, Department of Environmental Health Sciences, Johns HopkinsUniversity, Baltimore, MD, 21205. Phone: (410) 955-4712. Fax: (410) 955-0116. E-mail: tkensler@jhsph.edu.#Dr. Fahey, as well as Johns Hopkins University, owns stock in Brassica Protection Products, LLC (BPP), a company whose mission isto develop chemoprotective food products and which sells broccoli sprouts. Dr. Fahey is an unpaid scientific consultant to BPP and hisfounder’s stock is subject to certain restrictions under University policy. The terms of this arrangement are being managed by JohnsHopkins University in accordance with its conflict of interest policies.
NIH Public Access
Author Manuscript
Chem Res Toxicol
. Author manuscript; available in PMC 2008 September 8.
Published in final edited form as:
Chem Res Toxicol
. 2007 April ; 20(4): 572–576. doi:10.1021/tx7000459.
N I  H -P A A  u t  h  or M an u s  c r i   p t  N I  H -P A A  u t  h  or M an u s  c r i   p t  N I  H -P A A  u t  h  or M an u s  c r i   p t  
 
Chemical Classes of Inducers
A great deal of effort has gone into the identification of structural moieties that inducechemoprotective enzymes and this structure-inducer potency data has been reviewed recently(10). Some of these protective-enzyme inducing phytochemicals, for example the glucosinolatederivative isothiocyanates and indoles, are found exclusively in broccoli and its close relatives.Others, such as the carotenoids, flavonoids, and chlorophyll, are of almost universalprovenance in green plants, with antioxidant, anti-inflammatory, anti-allergic,anticarcinogenic, and even antiviral activities having been ascribed to them (11,12). The majorstructural classes of naturally occurring inducers are summarized in Table 1. Most, but not allof them are phytochemicals. In our work, we have given special attention to the glucosinolatesand isothiocyanates from
 Brassica
vegetables such as cauliflower, Brussels sprouts, broccoli,and cabbage (13), to chlorophyllin (14–16) which can be derived from extracts of virtually anygreen plant, and to certain flavonoids (17) and triterpenoids (18). To identify and isolateinducers of chemoprotective enzymes, we and others have utilized the Prochaska microtiterplate NQO1 assay (which measures the induction of quinone reductase, one of thequintessential detoxification enzymes in the adaptive stress response (19,20). Guided by thisapproach, sulforaphane was isolated from broccoli in 1992 (21). The use of this bioassay hasalso guided studies of structure-activity relationships, the synthesis of analogues, elucidationof mechanisms, and assessment of potencies (22).
Mode of Action
Multiple defense systems have evolved in all multicellular organisms in order to ensureprotection against the toxic effects of the plethora of endogenous and exogenous oxidants andelectrophiles to which they are exposed. DNA damage resulting from these reactiveintermediates is an integral component of carcinogenesis, and strategies to reduce the burdenof damage to the genome have been clearly shown to prevent cancer development in animals(6,23). There exist several signaling pathways that evoke an adaptive response to the stresselicited by oxidants and electrophiles. A key pathway is that of Keap1-Nrf2-ARE signaling.In mice, disruption of this pathway dramatically alters the response of animals to chemicalcarcinogens, oxidants, inflammatory states, and other toxins targeting a variety of organs (7).Induction of this cytoprotective response system requires at least three essential components:(i) Antioxidant Response Elements (AREs) (24), upstream regulatory sequences present oneach responsive gene either in single or multiple copies; (ii) Nrf2, the principal transcriptionfactor that heterodimerizes with members of the small Maf family of transcription factors,binds to the ARE, and recruits the general transcriptional machinery for expression of ARE-regulated genes (25); (iii) Keap1, a cytosolic repressor protein that binds to Nrf2, retains it inthe cytoplasm, and promotes its proteasomal degradation (26). Inducers react with thiols inKeap1 at rates that are closely related to their potencies, leading to disruption of the Nrf2-Keap1 complex and nuclear accumulation of Nrf2 (27). The molecular features of this pathwayhave been recently reviewed in
Chemical Research in Toxicology
(28) and elsewhere (29), butit is already clear that phytochemicals such as sulforaphane directly interact with Keap1 totrigger a gene expression response (30).
Sources and Delivery of Cytoprotective Agents
Extracts of plants and isolated natural products from those plants exhibit a bewildering arrayof activities in bioassays and in animal models, and we are beginning to see such productssubjected to rigorous scrutiny in human volunteers. The induction of the cytoprotectiveenzymes has now been widely studied in cell culture and in animal models, and clinicalinvestigations are beginning to unfold (31). A wealth of information exists on cell-based assaysthat have good predictive value, at least to the stage of animal models of carcinogenesis. Aninteresting conclusion that can be drawn from this body of literature: There are a number of 
Fahey and KenslerPage 2
Chem Res Toxicol
. Author manuscript; available in PMC 2008 September 8.
N I  H -P A A  u t  h  or M an u s  c r i   p t  N I  H -P A A  u t  h  or M an u s  c r i   p t  N I  H -P A A  u t  h  or M an u s  c r i   p t  
 
potent, naturally occurring inducers of protective enzymes that also have low cytotoxicity,(
e.g.,
sulforaphane, resveratrol, pinostrobin, and EGCG). By our calculations, consumption of a serving or two of foods like broccoli sprouts (sulforaphane glucosinolate) or Thai ginger(pinostrobin) can be expected to be effective in inducing the chemoprotective enzyme response(17,32). For example, we recently conducted a clinical study in China using standardized dosesof broccoli sprout derived sulforaphane glucosinolate, and observed a dose-dependantreduction in markers of DNA damage (33). Levels of sulforaphane were calibrated to beachievable by dietary means. However, the consumption of quite high levels of the plants inwhich some of these compounds are found would be necessary to achieve what can bereasonably expected to be a protective, but not toxic level of active phytochemical. Whencalculations are made of the quantity of certain other potentially protective phytochemicals(
e.g.,
from wasabi or mustard), achieving an efficacious dose does not appear to be possible(or advisable) by dietary means.This presents a conundrum. Should persons at increased risk for particular cancers beencouraged to take supplements or food products enriched in potentially protectivecompounds? There is tremendous variation in the inducer potency of these compounds (Figure1). The consumption of a varied diet rich in potentially protective foods (Table 2) has clearattraction from the perspective of safety (low risk of overdosing or toxicity). On the other hand,the evolution of modern humans’ diet has been phenomenally rapid over the past fewgenerations (Figure 2) and an argument can be made that dietary supplementation may be calledfor if we are to achieve a more uniformly enhanced longevity across populations – anincompletely understood phenomenon which is now enjoyed by a fraction of individuals inWestern and other populations. Thus, we have also approached the issue of substances whichhave a very high safety index (
e.g.
, one can consume a lot of it without adverse effects) butrelatively low inducer potency, in an alternative fashion. We have examined compounds suchas chlorophyllin and chlorophyll (14–16) and foods such as honey (17) which have very longand clear records of safe consumption, but do not have particularly high potency as inducersof chemoprotective enzymes. Dogmatic approaches to protection frequently assume that onewould need to attain high levels of protection in order for there to be value in this strategy.Whereas this may be the only paradigm which permits us to measure risk reduction usingcurrently available tools, it also embodies the flawed logic that the only good risk reduction isa large risk reduction. Consider the following examples.In the context of the amount of sugar consumed in economically advantaged countries (194 g/ d of sugar is eaten by Americans) (34), we have determined that replacing a small fraction of this quantity with honey, might well yield small but significant cancer protective results inaddition to all of the other obvious benefits of replacing white sugar or high fructose corn syrupwith honey (17). A similar argument can be made for chlorophyll and some of the accessoryplant pigments (
e.g.,
carotenoids and xanthophylls) which are present in essentially all higherplants (fruits and vegetables), yet are only moderately to slightly potent inducers in the assaysystems that have been employed (16). Thus, the additive effect of a multitude of weak tomoderately potent inducers for people eating 5 or more servings a day of fruits and vegetablesmay be highly significant. Reducing the risk of cancer by only 1%, in this country alone, wouldequate to saving about 5,600 lives. If one assumes that these lives saved resulted in an averageof 15 years of additional life, this would account for 82,500 life-years saved (a frequently usedmetric) per year of dietary change or intervention. Thus although it may be next to impossibleto measure as small a risk reduction as 1 or 2% (placebo-controlled clinical trials with a tumorendpoint would be impossible to conduct), the significance of even this magnitude of preventive effect should not be lost on policy makers. The risks in this example are essentiallynon-existent.
Fahey and KenslerPage 3
Chem Res Toxicol
. Author manuscript; available in PMC 2008 September 8.
N I  H -P A A  u t  h  or M an u s  c r i   p t  N I  H -P A A  u t  h  or M an u s  c r i   p t  N I  H -P A A  u t  h  or M an u s  c r i   p t  
of 00

Leave a Comment

You must be to leave a comment.
Submit
Characters: ...
You must be to leave a comment.
Submit
Characters: ...