Journal of Food and Nutrition Sciences 2018 (Nov 26); 6 (5): 129–134 ~ FULL TEXT
Hong Wang, Bruce Paul Daggy, Jamie Francine McManus, Les G Wong
Research & Development,
Why was this study done? |
There is a great deal of interest in understanding more about preserving telomere length; telomeres are the protective caps on the end of our DNA strands. It is known that diets and specific nutrients seem to have a protective effect, especially multivitamins, omega 3, vitamin D and polyphenols. This study compared telomere lengths over the age spectrum from ages 30–79 (by decades) between a population of heavy supplement users (Shaklee) and a healthy age-matched control group of non-smoking, non-obese San Francisco Bay area population.
What This Study Found
This study showed that long-term multiple supplant users had longer telomere lengths in their 50s, 60s and 70s as compared to the controls. The results suggest that heavy dietary supplementation may slow the shortening of telomeres that occurs with aging. Further research is needed to understand the mechanisms of action of specific nutrients and telomere protection.
Telomeres are DNA-protein complexes that cap chromosomal ends to promote chromosomal stability. Telomere length has been associated with diet and nutrition and inversely associated with mortality, life stress, and diseases. The objective of the current study was to explore the effect of long-term intake of multiple dietary supplements on telomere length in adults compared with age-matched controls. Salivary telomere lengths were determined from a group of heavy supplement users (HSU group; n=81; 20 males and 61 females age 30–79), the majority of whom took > 12 different supplements daily at least 4 days per week for at least 5 years, compared to a population of relatively healthy, non-smoking, age-matched subjects in the San Francisco Bay Area (control group; n=323; 146 males and 177 females) who took ≤ 3 different supplements daily.
Telomere lengths were measured by quantitative PCR to determine the telomere-to-single copy gene (T/S) ratio and were compared between groups that were age-matched by decade from 30 to 79. There was an age effect (p≤0.05) and a significant difference between groups (p≤0.001). The HSU group had significantly longer telomere length in the 50s, 60s, and 70s age groups (p ≤0.01, 0.001, 0.01 respectively). Routine clinical blood chemistry biomarkers (glucose, lipid profiles, liver enzymes, renal function and CRP) in both groups were all within the normal ranges. The results of this cross-sectional study suggest that heavy dietary supplementation may attenuate telomere shortening compared to an age-matched control group. Longitudinal studies are needed to further explore the potential association between nutritional supplementation, healthy aging and the reduced rate of telomere shortening.
Keywords Telomere Length, Telomere Shortening, Dietary Supplements, Nutrients, Diets, Oxidative Stress, Inflammation
From the FULL TEXT Article:
Telomeres are DNA-protein complexes that cap
chromosomal ends, keeping these ends from fusing with each
other and from being recognized as a double strand break by
DNA repair proteins in the cell. During cell division, DNA
replication cannot fully copy to the extreme ends of the linear
chromosome leading to telomere attrition during cell
division. Although telomerase can add telomeric repeat
sequences to the end of chromosome, its levels and activity
are limited in many human cells leading to the telomere
shortening throughout the life span.  Telomere length has
been associated with lifestyle, diet and nutrition and
inversely associated with aging, stress and diseases. [2–6]
Short telomeres are also associated with mortality risk. 
Leukocyte telomere length is correlated with telomere length
of various tissues and the systemic influence on telomere
maintenance and has been proposed as a surrogate biomarker [1, 7] which may reflect physiological age and health status.
Previously a cross-sectional study showed that long-term
multiple dietary supplement usage was associated with
significantly improved nutritional status and multiple
measures of health status.  Other clinical studies had
shown that multivitamin use in women  and
micronutrients consumption in a Korean middle-aged
population  were associated with longer telomere length.
Increasing blood levels of carotenoids were shown to be
associated with longer telomere length in some US
populations.  Other nutrients such as vitamin C, vitamin
D, and omega-3 fatty acids levels were also associated with
longer telomeres or attenuation of telomere shortening. [12–15] The blood levels of these nutrients are directly related to
dietary intake from food or dietary supplements. All these
studies suggest a potential impact of dietary supplement use
on telomere length. The objective of the current crosssectional
study was to explore the effect of long-term
multiple dietary supplementation intake on telomere length
compared to age-matched controls.
Two groups of participants (male and female, age between
30–79 years old) were recruited.
1) Heavy Supplement Users (HSU) group: supplement users who had been product consumers of a dietary supplement manufacturer and distributor (Shaklee Corporation, Pleasanton, CA) who took at least 5 different supplements 4–7 times weekly for at least 5 years. Participants in HSU group were from all regions of the mainland US.
2) Control (CON) group: relatively healthy non-smokers who took no more than 3 different supplements daily. CON participants were living in San Francisco Bay Area.
1) HSU group: recruited at a conference held by the
supplement distributor in Pleasanton, CA.
2) CON group: recruited through agreements with local
companies who agreed to advertise the study and allow
their employees to participate at the workplace.
Recruitment was via email flyers sent by each
employer’s Human Resource department. Additional
residents in the San Francisco Bay area were recruited
through newspaper and radio advertisements, fliers and
1) HSU group: men and women aged between 30–79
years old residing in mainland USA who used at least 5
different supplements 4–7 times weekly for at least 5
years and were willing to sign the informed consent
form. Dietary supplement usage information in this
group was assessed via online survey questionnaires
completed by 74% of HSU participants. Common
supplements used include single- and multi-vitamins
and minerals, fish oil, and supplements with various
combinations of herbal extracts. Within the returned
responses for supplements taken 4–7 times weekly,
98% of respondents were taking a multi-vitamin,
multi-mineral supplement, 94% were taking single
vitamins or a mixed carotenoid supplement, 55% were
taking mineral supplements, 97% were taking
supplements containing omega-3 fatty acids and 91%
were taking a resveratrol and muscadine polyphenols
2) CON group: relatively healthy men and women aged
between 30–79 years old residing in SF Bay Area who
were English-speaking and willing to sign the informed
consent form and keep healthy behaviors stable for one
1) HSU group – no exclusion for participation.
2) CON group – those with health conditions and/or
taking medications that would affect telomere length
except for those in aged 60–70s years old with mild
conditions of aging and common medications (statins
or antihypertensives). Anyone who used tobacco or
recreational drugs, was obese (BMI >35 kg/m2), took
more than 3 daily supplements, or took > 1g/day
supplemental omega-3 fatty acids from fish oil was
excluded from the CON group.
Sample collection and processing as well as telomere
length assay and data collection were performed by Telomere
Diagnostics, Menlo Park, CA. The study protocol was
reviewed and approved by Institutional Review Board
(Ethical & Independent Review Services, Corte Madera,
CA). All participants provided informed consent.
Telomere Length Test
Quantitative PCR was used to measure average telomere
length per genome (i.e. telomere-to-single copy gene (T/S)
ratio) in saliva cellular DNA. Saliva was used because of its
ease of collection and storage. Saliva DNA sample was
shown to be a viable alternative to blood samples  and
the main source of the DNA comes from saliva leukocytes.  Saliva samples were collected with Oragene DNA collection kit (Ontario, Canada).
Blood Chemistry Test
Fasting blood samples were assayed for routine blood
chemistry tests by LabCorp Testing Laboratory (Burlington,
NC). Exclusion criteria also included missing, ineligible
participants’ biological specimens or samples where
hemolysis invalidated the reading.
Data Collection, Exclusion and Statistics Analysis
Telomere length was reported as a T/S ratio (telomere
signal normalized relative to a single copy gene signal).
Samples with T/S ratio >2.0 were excluded from the data set
due to potential artifact.  Exclusion criteria also included
missing or ineligible participants’ biological specimens or
where hemolysis invalidated the reading. After exclusion,
there were 323 subjects in the CON group and 81 in the HSU
group that were within the age from 30 to 79 years old
T/S ratios of the Heavy Supplement Users (HSU) group were
compared to those in the age-matched CON group by using
linear regression and two-way ANOVA with multiplecomparisons
(Bonferroni post-hoc test) using Graphpad
Prizm 6.0. P-values ≤ 0.05 was considered statistically
The results of the salivary telomere length of the
participants from 2 groups are shown in Figure 2.
plots of each group are shown in a) and b). Then, the data
were grouped based on their age in the decades from 30s to
70s as shown in c). Linear regression was used to plot the
graph for each group (CON: Y = –0.004869*X + 1.357; HSU:
Y = –0.002107*X + 1.375). There is no significant difference
between the slopes of the lines. When using two-way
ANOVA with multiple comparisons for between-group
comparisons by decade from 30s to 70s, there was an age
effect (p ≤ 0.05) and a significant difference between groups
(p≤0.001) with no significant interaction (p = 0.5488). The
HSU group had significantly longer telomere length in the
age group at 50s, 60s and 70s (p ≤ 0.01, 0.001, 0.01
The blood chemistry results from two group of participants
are shown in Table 1. All the means were within normal
ranges in both groups.
Telomere length has been shown to associate with aging,
diseases, life style, stress, diet and nutrition. [2–6] Telomere
length is heritable but can be modified by various
environmental factors and health status. Key factors affecting
telomere length are oxidative stress, inflammation and DNA
integrity.  Diet and nutrition including several nutrients or
phytonutrients which have been associated with attenuation
of telomere shortening can be categorized based on these
Oxidative stress and inflammation play a significant role in
aging and diseases that contribute to tissue injury with
cellular and DNA damage. Many antioxidants and antiinflammatory
agents can attenuate telomere shortening. 
Oxidative stress can cause oxidative base modifications and
DNA breaks.  Oxidative damage is repaired less well in
telomeric DNA and oxidative stress accelerates telomere loss,
whereas antioxidants decelerate it.  Nutrients such as
carotenoids, vitamin C and vitamin E are well-known
antioxidants and play a significant role in the body to protect
against oxidative stress. These nutrients have been shown to
associate with longer telomere length in three different
population group. [9–12]
Inflammation increases the rate of immune cell turnover
and replication, which would contribute to telomere attrition.
In addition, activated immune cells during inflammation
generate reactive oxygen and nitrogen species that cause
oxidative stress-induced DNA damage.  Infectioninduced
chronic inflammation in patients was associated with
shortened telomere length [22, 23] illustrating the significant
role of inflammation. Therefore, certain nutrients that can
counter inflammation in the body could potentially attenuate
its deleterious effects on telomere shortening. In addition to
the enhancement of calcium absorption, vitamin D has antiinflammatory
actions via multiple pathways. Serum vitamin
D levels were negatively associated with C-reactive protein
(CRP), an inflammatory biomarker and were positively
associated telomere length. [14, 24, 25]
mediators generated in the body originate from cell
membrane lipids. Dietary lipids have direct impact on the
types and amount of fatty acids incorporated into cell
membranes. Omega-6 (n-6) fatty acids such as arachidonic
acid are the precursors that generate pro-inflammatory
prostaglandins and leukotrienes, in contrast to omega-3 (n-3)
fatty acids, including eicosapentaenoic acid (EPA) and
docosahexaenoic acid (DHA), which are precursors to many
anti-inflammatory prostaglandins and resolvins. There was
an inverse association of omega-3 fatty acids levels with the
rate of telomere shortening over 5 years in coronary heart
disease patients in a longitudinal study.  Omega-3
supplementation in healthy overweight middle aged and
older adults for 4 months reduced oxidative stress, and the
lower n-6:n-3 ratio from this supplementation was associated
with an increase in telomere length.  Other food
components that have anti-inflammatory effects based on
negative dietary inflammatory index scores can also
contribute to effects on telomere length. [26, 27]
Telomere length is influenced by DNA integrity and DNA
and histone methylation.  Folate is essential for the
production and maintenance of DNA and cell cycle
progression through methylation. However, when there is
loss of DNA and histone methylation, epigenetic regulation
is lost resulting a more elongated telomere than normal. 
Therefore, the effects of folate on telomere length depend in
part on cellular status. While consumption of folate based on
a food frequency questionnaire was positively associated
with telomere length in middle-aged Korean adults < 50
years old and not older , leucocyte telomere length and
plasma folate status had been studied in a group of Italian
men and  and a larger study in Framingham Offspring
However, the results from these two studies were
not consistent with each other with the Italian men study
showing the plasma folate level had a U-shaped relationship
with telomere length but was shown inversely associated
with telomere length only at very high plasma folate level in
the Framingham cohort study. Therefore, the actual
relationship between folate status and telomere length
Besides single nutrients, certain healthy diet patterns or
food groups were also associated with telomere length. A
recent systemic review supported the findings that adherence
to Mediterranean diet (MD) was associated with longer
telomere length; fruits and vegetables are the two food
groups which showed positive association.  The MD is
characterized by: a high intake of vegetables, fruits, legumes
and grains; a high intake of olive oil and low in saturated fat;
a moderately high intake of fish; a low intake of dairy
products, meat and poultry; and a regular intake of wine.
There are well-established protective effects of the MD on
oxidative stress and chronic inflammation which may explain
the favorable influence on telomere length.  Key
phytonutrients from the diet are polyphenols, found in fruits
and vegetables, olive oil, coffee and wine. Polyphenol intake
from a MD was shown to reduce the inflammation and
improve cardiovascular and mortality risk. [32–34] A recent
study also showed that higher adherence to a healthy diet
using Mediterranean Diet scores, Healthy Eating Index–2010
scores, and Dietary Approaches to Stopping Hypertension
(DASH) scores as the diet quality indices were each
associated with longer telomere length in women. 
In the current cross-sectional study, the salivary telomere
lengths from age-matched long term heavy supplement users
were compared to those of relatively healthy controls, who
did not have conditions or behavior such as smoking that
may affect the telomere length. The results showed that there
are an age effect and a significant difference between groups.
The HSU group had longer telomeres than the CON group
from age 50s to 70s. Both groups had normal blood
chemistry including CRP within the normal range. The
results seem to reflect a previous study, which showed that
long-term multiple dietary supplement usage was associated
with significantly improved health and nutritional status. 
Within the returned responses of the supplement use
questionnaires from the HSU participants for supplements
taking 4–7 times weekly, 98% were taking a multi-vitamin,
multi-mineral supplement, 94% were taking single vitamins
or a multi-carotenoid supplement, 55% were taking mineral
supplements, 97% were taking supplements containing
omega-3 fatty acids and 91% were taking a resveratrol and
muscadine polyphenols supplement. Similar to other studies
of diet pattern’s association with telomere length, it is
difficult to pinpoint exact mechanisms or contributions of the
individual components’ potential effects and interactions in
this study, particularly in the absence of measures of nutrient
status for these subjects.
This study showed that long-term multi-supplement users had longer telomere length in their 50s, 60s and 70s compared to controls. The results suggest that heavy dietary supplementation may attenuate telomere shortening and is consistent with a previous study’s findings that long-term multiple dietary supplement usage was associated with significantly improved health and nutritional status. Based on the published literature and putative mechanisms, supplements containing vitamins C, D and E, carotenoids, omega-3 fatty acids and polyphenols may all contribute to the effects on telomere length. Further research, including prospective longitudinal studies, is needed to have a better understanding of the active nutrients and their mechanisms of action.
Conflict of Interest Statement
The authors declare that they have no competing interests.
HSU: heavy supplement users;
T/S: telomere-to-single copy gene;
PCR: polymerase chain reaction;
ANOVA: analysis of variance;
CRP: C-reactive protein;
EPA: eicosapentaenoic acid;
DHA: docosahexaenoic acid;
MD: Mediterranean diet.
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