Standard Process sponsors these webinars to

provide healthcare information to practitioners

and to provide them the opportunity to hear
about the views, recommendations and
experiences of other practitioners.
Standard Process has not evaluated the legal
status of any products, services or
recommendations with respect to State or
Federal laws, including scope of
practice. Standard Process does not and
cannot accept responsibility for errors or
omissions or for any consequences from
applications of the information provided and
makes no warranty, expressed or implied, with
respect to the information provided.

Antioxidants:
A New Understanding

Kerry Bone
Co-Founder and Director R&D
MediHerb
Adjunct Associate Professor
University of New England, Australia

Topics Covered
Basic redox chemistry and definitions
Herbal (phytochemical) antioxidants: important
considerations and controversies
The essential aspects of the Nrf2/ARE pathway a new insight into clinically relevant antioxidant
protection
The clinical relevance of the pathway for health,
wellness and normal processes of aging
Herbs and phytochemicals shown to act via the
Nrf2/ARE pathway
Current and novel clinical applications informed
by this new understanding
3

Kerry Bone 2012

MH225

Basic Redox Chemistry

and Definitions
Redox (short for reduction-oxidation) describes
chemical reactions where the oxidation state of
component atoms changes
With oxidation (one half of
the redox reaction) there
is a loss of electrons or
an addition of oxygen
With reduction (the other
half of the redox reaction)
there is a gain of electrons
or an addition of hydrogen
In a redox reaction the oxidant (oxidizing agent)
is reduced and the reducing agent is oxidized
4

Basic Redox Chemistry

and Definitions
Hence there is potentially a flow of electrons
from the reducing agent to the oxidant
A chemical that is good at giving away electrons
is a powerful reducing agent
Similarly, a chemical that actively seeks electrons
is a powerful oxidising agent
Because electron flow is often involved,
chemicals involved in redox reactions can be
placed in an electrochemical series, depending
on their redox potentials (measured in volts)
The more negative the redox potential, the more
powerful the reducing agent
5

Basic Redox Chemistry

and Definitions
The more positive the potential, the more
powerful the oxidant
When a chemical is placed in this series,
anything with a more negative redox potential
can reduce it, anything with a more positive
potential can oxidize it
Hence, depending on the environment and what
it reacts with, an antioxidant (reducing) chemical
can become a pro-oxidant (oxidizing agent)
Moreover, any chemical entity involved in a redox
reaction has an oxidized form (that can act as
pro-oxidant) and a reduced form (that can act as
an antioxidant)
6

Kerry Bone 2012

MH225

Basic Redox Chemistry

and Definitions
In the human body, many key oxidizing agents
are based on oxygen and are called reactive
oxygen species (ROS)
ROS are produced as a normal product of cellular
metabolism
Many ROS have unpaired
electrons that need to find
a pair, such chemicals are
known as free radicals
Free radicals can affect
tissue by causing cross
linking of molecules and chain reactions
7

The Value of Antioxidants

One of the most critical functions of life is
the ability to manage oxidative stress1
Oxygen is a two-edged sword: it provides
energy for life but the by-products (ROS) are
corrosive to tissue, just as oxygen in the
atmosphere is corrosive to metal
The body is exposed to other pro-oxidants via
electromagnetic radiation and the things we
ingest or inhale
Hence, management of oxidative stress,
especially within the cell, is critical to its viability
ROS have been implicated in the etiology of a
range of health challenges
1

Maher J, Yamamoto M. Toxicol Appl Pharmacol 2010;

244(1): 4-15

Kerry Bone 2012

MH225

Controversies Over
Antioxidants
The oxidative stress theory of aging (and hence
the value of consuming antioxidant supplements)
has been challenged by many researchers, even
former advocates (see later in slide 13)
Antioxidants might become pro-oxidants in the
right environment: examples include vitamin C
and the negative clinical results for isolated betacarotene in smokers
The discovery of ROS signalling: demonstrated in
a variety of physiological processes including
glucose-stimulated insulin secretion and induction
of host defenses1
1

Zhang Q, Pi J, Woods CG et al. Toxicol Appl Pharmacol

2010; 244(1): 84-97
10

Herbal Antioxidants are

Misunderstood
A number of methods are used to assess the
antioxidant activity of plant-based foods and
therapeutic agents
These include ORAC (oxygen radical
absorbance capacity), TRAP (total radicaltrapping antioxidant parameter) and TEAC
(trolox equivalent antioxidant capacity)
However, they are all test tube methods that
probably have little bearing on what actually
happens in the body after the plant is
ingested
11

Herbal Antioxidants are

Misunderstood
Specifically, it has been shown that often only a
small fraction of the antioxidant phytochemicals
that contribute to ORAC etc are absorbed as such
Or what is absorbed exerts relatively negligible
activity directly quenching ROS in body fluids
This is supported by a lack of co-ordination
between ORAC levels and the ability to control
oxidative stress in the body
In any case, bodily fluids already contain
significant levels of an active antioxidant
molecule: uric acid
Finley JW, Kong AN, Hintze KJ et al. J Agric Food Chem 2011; 59(13): 6837-6846
Niki E. Curr Pharm Biotechnol 2010; 11(8): 801-809
Cheli F, Baldi A. J Food Sci 2011; 76(9): R197-R205
12

Kerry Bone 2012

MH225

ORAC Values

13

The Oxidative Stress

Theory of Aging
This was first proposed by Harman
in 1956: ROS inflict indiscriminate
oxidate damage to cells that
cannot be completely neutralized
by antioxidants and/or repaired
Damage then accumulates with time aging
Despite the intuitive logic and vast support for
this theory, a causal link between oxidative
stress and the rate of aging has not been clearly
established1
Many workers now question its validity
1

Lewis KN, Mele J, Hayes JD et al. Integr Comp Biol 2010;

50(5): 829-843

14

A New Perspective
However, a common denominator exists in
several studies: cells from long-living animals
show greater levels of cytoprotection
They are more resistant to oxidative and
chemical insults
This strong relationship between species
longevity and cellular resistance to oxidative
insults has been reported in a wide variety of
species across the animal kingdom (see later
in slide 23)
15

Kerry Bone 2012

MH225

Important
Cellular
Protector
Pathways

16

The Nrf2/ARE Pathway

Otherwise known as the multiorgan protector
pathway, this is a basic cellular survival
mechanism
A primordial pathway, fundamental to all
animals cells, it describes a targeted approach
to antioxidant protection in the cell
The Nrf2/ARE pathway is a dynamic response
induced by oxidative/chemical stress on the cell
that is fundamentally new and novel
It is a switch-on, switch-off mechanism, and
most of the priming agents studied to date that
facilitate its responsivity are natural plant
chemicals
17

The Nrf2/ARE Pathway

Phytochemicals exerting a priming effect on the
NRf2/ARE pathway are hormetic and represent
an indirect way to safely provide antioxidant
protection for the cell
And the priming of this pathway brings much
more than antioxidant effects

18

Kerry Bone 2012

MH225

The Nrf2/ARE Pathway

19

The Nrf2/ARE Pathway

20

Functions of the
Nrf2/ARE Pathway
As noted, Nrf2 is expressed in all tissues, but
levels vary, with higher levels in key detoxifying
organs: the liver and kidneys
Activation of Nrf2 activates
more than 200 genes
Nrf2 activation enhances
DNA repair, heme metabolism,
efflux transport of toxins and
glutathione synthesis
It activates detoxification, stabilizes proteins,
strengthens cellular integrity and moderates the
inflammatory response process
21

Kerry Bone 2012

MH225

Functions of the
Nrf2/ARE Pathway
Key antioxidant enzymes are induced, including
catalase, superoxide dismutase (SOD),
thioredoxin, peroxiredoxin, sulfiredoxin, ferritin,
metallothionein and haem oxygenase 1 (HO-1)
Key phase II enzymes are also induced, including
glutathione S-transferases (GSTs) and NAD(P)H:
quinone oxidoreductase 1 (NQO1)
These functions likely represent only a fraction of
the true function of Nrf2 activity, some of which
are only beginning to be understood
Maher J, Yamamoto M. Toxicol Appl Pharmacol 2010; 244(1): 4-15
Lewis KN, Mele J, Hayes JD et al. Integr Comp Biol 2010; 50(5): 829-843
Itoh K, Mimura J, Yamamoto M. Antioxid Redox Signal 2010; 13(11): 1665-1678
22

The Nrf2/ARE Pathway

in Health and Wellness
Current research has identified several important
involvements of the Nrf2/ARE pathway in
supporting and maintaining health:

healthy aging and longevity

benefits in health challenges
involving natural oxidative stress
and normal inflammation
potential benefit in health
challenges resulting from accumulated
environmental challenges
23

The Nrf2/ARE Pathway

and Longevity

Lewis KN, Mele J, Hayes JD et al. Integr Comp Biol 2010; 50(5): 829-843
24

Kerry Bone 2012

MH225

The Nrf2/ARE Pathway

and Neuroprotection
Beneficial effects of Nrf2/ARE
priming/activation have been
suggested in neurological
health1,2,3,4
HO-1 is a known neuroprotective
heat shock protein/enzyme that,
for example, supports brain
function5

25

The Nrf2/ARE Pathway

and Neuroprotection
References for previous slide
1 Negi G, Kumar A, Joshi RP et al. Oxidative stress and Nrf2 in the

pathophysiology of diabetic neuropathy: old perspective with a new

angle. Biochem Biophys Res Commun 2011; 408(1): 1-5
Calkins MJ, Johnson DA, Townsend JA et al. The Nrf2/ARE pathway as
a potential therapeutic target in neurodegenerative disease. Antioxid
Redox Signal 2009; 11(3): 497-508
Cuadrado A, Moreno-Murciano P, Pedraza-Chaverri J. The transcription
factor Nrf2 as a new therapeutic target in Parkinson's disease. Expert
Opin Ther Targets 2009; 13(3): 319-329
Plafker SM. Oxidative stress and the ubiquitin proteolytic system in
age-related macular degeneration. Adv Exp Med Biol 2010; 664:
447-456
Jazwa A, Cuadrado A. Targeting heme oxygenase-1 for
neuroprotection and neuroinflammation in neurodegenerative
diseases. Curr Drug Targets 2010; 11(12): 1517-1531
26

The Nrf2/ARE Pathway

and Brain Health
In one study, cells from biopsy of the
olfactory mucosa had lower glutathione
levels and detoxifying capacity in patients
with brain challenges compared with those
from healthy control donors
These cells from the challenged patients
were also in a state of oxidative stress due to
higher levels of hydrogen peroxide
Significantly, activation of the Nrf2/ARE
pathway restored those defective cells to
normal functioning
Cook AL et al. PLoS One 2011; 6(7): e21907

Kerry Bone 2012

MH225

27

NRF2/ARE and Other

Body Systems
Beneficial effects of Nrf2/ARE priming have also
been suggested for:

lung support
cardiovascular support
immune system support
blood glucose support

Singh S, Vrishni S, Singh BK et al. Nrf2-ARE stress response mechanism: a

control point in oxidative stress-mediated dysfunctions and chronic
inflammatory diseases. Free Radic Res 2010; 44(11): 1267-1288
28

Key Herbs/Phytochemicals
with Nrf2/ARE Activity
EGCG from Green Tea is also very active (in vitro
and in vivo - 20 mg/kg, oral)
Curcumin from Turmeric is
positive in many studies
(in vitro and in vivo, oral)
Resveratrol shows marked
in vitro and in vivo activities
and was also positive in an
human trial (see later)
Surh YJ, Kundu JK, Na HK. Nrf2 as a master redox switch in turning on the
cellular signaling involved in the induction of cytoprotective genes by some
chemopreventive phytochemicals. Planta Med 2008; 74(13): 1526-1539
Hur W, Gray NS. Small molecule modulators of antioxidant response pathway.
Curr Opin Chem Biol 2011; 15(1): 162-173
29

Key Herbs/Phytochemicals
with Nrf2/ARE Activity
Carnosol from Rosemary is another wellstudied phytochemical
Various phytochemicals from Garlic are
active in vitro
Several flavonoids exhibit in vitro activity,
but this may not translate to clinical
relevance because of their low oral
bioavailability
Surh YJ, Kundu JK, Na HK. Nrf2 as a master redox switch in turning on the
cellular signaling involved in the induction of cytoprotective genes by some
chemopreventive phytochemicals. Planta Med 2008; 74(13): 1526-1539
Hur W, Gray NS. Small molecule modulators of antioxidant response pathway.
Curr Opin Chem Biol 2011; 15(1): 162-173
30

Kerry Bone 2012

MH225

10

Rosemary
Rosemary contains the antioxidant
molecules carnosol and carnosic acid
Both of these are now understood to be potent
primers of the Nrf2/ARE pathway
For example both compounds have demonstrated
potential neuroprotective activity mediated by
this pathway, which reflects on its traditional use
for memory1,2
Recently a single 750 mg dose of Rosemary
improved speed of memory and alertness in
healthy older adults1
1
2
3

Kosaka K et al. J Biochem 2010; 147(1): 73-81

Martin D et al. Biol Chem 2004; 279(10): 8919-8929
Pengelly A et al. J Med Food 2012; 15(1): 10-17

31

32

Turmeric/Curcumin
Multiple in vivo studies have demonstrated
the activities of curcumin and Turmeric1
The Nrf2/ARE pathway is thought to be an
important basis for these effects
For example, the epigenetic silencing of Nrf2
during the progression of prostate challenge
in a mouse model was reversed by
curcumin2
1
2

Aggarwal BB, Surh YJ, Shisodia S (eds). Adv Exptl Med Biol 2007; 595;
1-489
Khor TO, Huang Y, Wu TY et al. Biochem Pharmacol 2011; 82(9): 1073-1078
33

Kerry Bone 2012

MH225

11

Turmeric/Curcumin
In other relevant models, chemically induced
oxidative stress was reduced by curcumin in
vivo in parallel with Nrf2/ARE upregulation1
Brain and liver were supported in vivo by
curcumin through Nrf2-mediated induction of
HO-12,3
Dietary curcumin led to increased Nrf2
protein levels and enhanced ARE binding in
liver and lungs of mice4
1
2
3
4

Charoensuk L et al. Int J Parasitol 2011; 41(6): 615-626

Yang C et al. Brain Res 2009; 1282: 133-141
Farombi EO et al. Food Chem Toxicol 2008; 46(4): 1279-1287
Garg R et al. Carcinogenesis 2008; 29(5): 1022-1032
34

35

36

Kerry Bone 2012

MH225

12

Resveratrol
Resveratrol is a highly active primer of the
Nrf2/ARE pathway
Favorable Nrf2-mediated activity has been
demonstrated in many body systems
including the endocrine, cardiovascular and
nervous systems
For example, the endothelial (circulatory)
protective effects of resveratrol against a
high-fat diet were largely diminished in Nrf2
knockout mice1
Hask G, Pacher P. Am J Physiol Heart Circ Physiol 2010; 299(1): H10-H12
37

Resveratrol
In a recent double blind, randomized,
crossover study, 10 normal, healthy men and
women were given a high-fat, highcarbohydrate (HFHC) meal (930 kcal) either
with a placebo or a product containing 100
mg of resveratrol from Polygonum
cuspidatum plus 75 mg of total polyphenols
from a grape extract1
Profound benefits were observed
1

Ghanim H, Sia CL, Korzeniewski K et al. J Clin Endocrinol Metab 2011; 96(5):
1409-1414
38

Resveratrol
DNA binding activity of Nrf2 in white cells
was significantly increased by 15039%
over baseline (p<0.05) at 3 hours after meal
and supplement intake, whereas meal
consumption in the placebo group resulted in
a significant reduction in Nrf2 binding
activity at 5 hours
This was associated with a significant
reduction of Keap-1 by 486% (p<0.05) in
the supplement group and a significant
increase by 6610% (p<0.05) in the
placebo group
39

Kerry Bone 2012

MH225

13

40

Ginkgo
Ginkgo is now well known as a powerful
clinical antioxidant. Its antioxidant properties
could play an important role in cellular
protection and appear to be mediated by Nrf2
In an uncontrolled trial conducted in 1995,
Ginkgo extract supported cellular health in
Russian workers1
More recently the same dose of extract
(120 mg/day) supported cellular health in
patients with thyroid challenges receiving
conventional therapies1
1
2

Emerit I et al. Radiat Res 1995; 144(2): 198-205

Dardano A et al. J Clin Endocrinol Metab 2007; 92(11):4286-4289
41

Ginkgo
There have been many trials of Ginkgo and brain
health in China and the Cochrane Collaboration
published a systematic review and metaanalysis1
While the review expressed concerns about the
quality of most trials, it found that Ginkgo was
associated with a significant increase in the
number of improved patients, based on
neurological function
Induction of HO-1 via Nrf2/ARE activation by
Ginkgo was suggested as a significant
mechanism for neuroprotection and support2,3
1
2
3

Zeng X et al. Cochrane Database Syst Rev 2005; (4): CD003691

Saleem S et al. Stroke 2008; 39(12): 3389-3396
Chen JS et al. Atherosclerosis 2011; 214(2): 301-309
42

Kerry Bone 2012

MH225

14

43

Korean Ginseng
Epidemiological and clinical studies support
the role of Korean Ginseng as a significant
promoter of cellular health
Emerging evidence suggests that priming of
the Nrf2/ARE pathway could be a key
mechanism involved1,2

1
2

Saw CL, Wu Q, Kong AN. Chin Med 2010; 5: 37

Park SH et al. Toxicology 2010; 278(1): 131-139

44

Vitanox
Each tablet contains:
Rosemary leaf 5:1 extract
from Rosmarinus officinalis leaf 1.0 g
Green Tea leaf 25:1 extract
from Camellia sinensis leaf 4.2 g
Containing catechins 83.35 mg
Turmeric rhizome 25:1 extract
from Curcuma longa rhizome 2.0 g
Containing curcuminoids 70.4 mg
Grape seed 120:1 extract
from Vitis vinifera seed 6.0 g
Containing procyanidins 42.5 mg
Dosage: 2 to 3 tablets per day

Kerry Bone 2012

MH225

200 mg
166.7 mg

80 mg

50 mg

45

15

HerbaVital
Each tablet contains:
Japanese Knot Weed root extract 100:1
from Polygonum cuspidatum root 8.0 g
Containing resveratrol 36 mg
Milk Thistle seed extract 70:1
from Silybum marianum seed 4.2 g
Containing flavanolignans calc. as silybin 48 mg
Korean Ginseng root extract 5:1
from Panax ginseng root 250 mg
Containing ginsenosides calculated as Rg1 and Rb1
Masson Pine bark extract 100:1
from Pinus massoniana bark 5.0 g
Containing proanthocyanidins 37.5 mg
Ginkgo leaf extract 50:1
from Ginkgo biloba leaf 1.5 g
Containing ginkgo flavonglycosides 7.2 mg
Containing ginkgolides and bilobalide 1.8 mg
Dosage: 2 to 4 tablets per day

80 mg
60 mg
50 mg
4.2 mg
50 mg
30 mg

46

A New Understanding
Providing clinically relevant, safe, targeted
antioxidant cover
Healthy aging and longevity protocols
Supporting detoxification pathways
Protecting against ANY physical or biological
stressor

47

A New Understanding
As key supporting agents for neurological
health across a range of health challenges
As key moderators in any chronic
musculoskeletal or immune system
challenges
As key supporting agents for the
cardiovascular system, especially arterial and
endothelial function

48

Kerry Bone 2012

MH225

16

A New Understanding
Countering the negative metabolic effects
of a high-fat and/or high-fried/food diet
Providing protective cover during
weight loss
For metabolic modulation
For lung support

49

A Cautionary Note
Nrf2 and its downstream genes are overexpressed in many experimental cancer cell
lines and human cancers, giving cancer cells
an advantage for survival and growth
Also Nrf2 is upregulated in cancer cells
resistant to chemotherapy and is thought to
be responsible for acquired chemoresistance1

Lau A, Villeneuve NF, Sun Z et al. Pharmacol Res 2008; 58(5-6): 262-270
50

A Cautionary Note
There are still many questions to be
answered
However, caution dictates that any known
Nrf2/ARE upregulating herbs should not be
taken at least 48 hours either side of each
chemotherapy or radiotherapy treatment

51

Kerry Bone 2012

MH225

17

A Final Reminder
The key herbs/phytochemicals identified by
current research that are most active at
priming the Nrf2/ARE pathway are found in:
Vitanox
Turmeric/curcumin, Rosemary and Green
Tea/ EGCG
HerbaVital
Resveratrol plus OPCs, Ginkgo and Korean
Ginseng
52

Questions

53

Kerry Bone 2012

MH225

18