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Pomegranate Juice and Extract Consumption Increases the Resistance to UVB-induced Erythema and Changes the Skin Microbiome in Healthy Women: a Randomized Controlled Trial

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In vitro and animal studies have demonstrated that topical application and oral consumption of pomegranate reduces UVB-induced skin damage. We therefore investigated if oral pomegranate consumption will reduce photodamage from UVB irradiation and alter the composition of the skin microbiota in a randomized controlled, parallel, three-arm, open label study. Seventy-four female participants (30–45 years) with Fitzpatrick skin type II-IV were randomly assigned (1:1:1) to 1000 mg of pomegranate extract (PomX), 8 oz of pomegranate juice (PomJ) or placebo for 12 weeks. Minimal erythema dose (MED) and melanin index were determined using a cutometer (mexameter probe). Skin microbiota was determined using 16S rRNA sequencing. The MED was significantly increased in the PomX and PomJ group compared to placebo. There was no significant difference on phylum, but on family and genus level bacterial composition of skin samples collected at baseline and after 12 week intervention showed significant differences between PomJ, PomX and placebo. Members of the Methylobacteriaceae family contain pigments absorbing UV irradiation and might contribute to UVB skin protection. However, we were not able to establish a direct correlation between increased MED and bacterial abundance. In summary daily oral pomegranate consumption may lead to enhanced protection from UV photodamage.
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Pomegranate Juice and Extract
Consumption Increases the
Resistance to UVB-induced
Erythema and Changes the Skin
Microbiome in Healthy Women: a
Randomized Controlled Trial
Susanne M. Henning
1, Jieping Yang1, Ru-Po Lee1, Jianjun Huang1, Mark Hsu1, Gail Thames1,
Irene Gilbuena1, Jianfeng Long2, Yunhui Xu1, Esther HaeIn Park1, Chi-Hong Tseng3,
Jenny Kim1,4, David Heber1 & Zhaoping Li1
In vitro and animal studies have demonstrated that topical application and oral consumption of
pomegranate reduces UVB-induced skin damage. We therefore investigated if oral pomegranate
consumption will reduce photodamage from UVB irradiation and alter the composition of the skin
microbiota in a randomized controlled, parallel, three-arm, open label study. Seventy-four female
participants (30–45 years) with Fitzpatrick skin type II-IV were randomly assigned (1:1:1) to 1000 mg
of pomegranate extract (PomX), 8 oz of pomegranate juice (PomJ) or placebo for 12 weeks. Minimal
erythema dose (MED) and melanin index were determined using a cutometer (mexameter probe).
Skin microbiota was determined using 16S rRNA sequencing. The MED was signicantly increased
in the PomX and PomJ group compared to placebo. There was no signicant dierence on phylum,
but on family and genus level bacterial composition of skin samples collected at baseline and after 12
week intervention showed signicant dierences between PomJ, PomX and placebo. Members of the
Methylobacteriaceae family contain pigments absorbing UV irradiation and might contribute to UVB
skin protection. However, we were not able to establish a direct correlation between increased MED
and bacterial abundance. In summary daily oral pomegranate consumption may lead to enhanced
protection from UV photodamage.
Exposure of human skin to UV radiation is a major factor for skin pathologies including erythema, inamma-
tion, degenerative age-related changes and cancer1. UV radiation is mostly composed of UVA (315–400 nm) and
UVB (290–320 nm). Overexposure of the skin to UVA and to a lesser extent to UVB leads to oxidative stress that
increases the generation of reactive oxygen species (ROS) causing lipid peroxidation of cell membranes, DNA
and protein damage to tissue, inammation and keratinocyte apoptosis24. ROS also trigger the expression of
matrix metalloproteinases (MMP), that degrade extracellular matrix such as collagen maintaining cell and skin
integrity3.
Pomegranate fruits have been used for centuries in ancient cultures for its medicinal purposes5. e health
benet of pomegranate has been attributed to the content of hydrolysable tannins (ellagitannins) including
punicalagins and ellagic acid (EA) as well as anthocyanins and other polyphenols found in pomegranate extract
1Center for Human Nutrition, David Geen School of Medicine, Department of Medicine, Los Angeles, CA, 90095,
USA. 2Department of Clinical Nutrition, 2nd Xiangya Hospital of Central South University, Changsha, Hunan, 410011,
China. 3Department of Statistics Core, David Geen School of Medicine, University of California Los Angeles, Los
Angeles, CA, 90095, USA. 4Division of Dermatology, David Geen School of Medicine, University of California Los
Angeles, Los Angeles, CA, 90095, USA. Correspondence and requests for materials should be addressed to S.M.H.
(email: shenning@mednet.ucla.edu)
Received: 19 June 2018
Accepted: 20 September 2019
Published: xx xx xxxx
OPEN
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and juice6. Although POM ellagitannins are highly bioactive in vitro, they are not absorbed intact in the small
intestine and undergo partial hydrolysis to form EA7. Ellagitannins and EA remaining in the large intestine are
further metabolized to urolithins A –D by the microbiota in the large intestine and are absorbed into the blood
stream810. erefore, pomegranate properties could be mediated by the metabolites produced in the intestine in
addition to the original phenolic compounds present in the food matrix. Due to the sugar content of PomJ, which
might be of health concern, we included both PomJ and PomX intervention in this investigation.
Several in vitro and animal studies provide evidence that either topical application or oral consumption of
pomegranate juice (PomJ) or extract (PomX) or EA reduce damage from UVB irradiation1117. One human
study demonstrated that oral consumption of an EA rich pomegranate extract in healthy women was asso-
ciated with a protective eect on slight sunburn caused by UV irradiation even at a low dose resulting in a
decrease in pigmentation17. In vitro antioxidant and anti-inammatory activity of ellagitannins and EA have
been widely demonstrated1820, while oral ingestion of pomegranate in humans has been less investigated. Some
human studies provide mechanistic evidence that pomegranate ingestion leads to an increase in antioxidant and
anti-inammatory activity9,21.
Inammation caused by UV radiation activates various matrix-degrading matrix metalloproteases (MMPs),
which leads to collagen degradation and cellular apoptosis. MMP-1, especially, was the main endogenous factor
that degraded dermal collagen in the process of human skin senility22.
Previously published ndings from our laboratory demonstrated that pomegranate extract inhibited in vitro
growth of bacteria involved in the pathogenesis of acne including Propionibacterium acnes, Propionibacterium
granulosum, Staphylococcus aureus and Staphylococcus epidermidis23. Although the concentration of circulating
Pom ellagitannin metabolites is very low, there is a possibility that Pom ellagitannin metabolites in the skin might
alter the composition of the skin microbiota8. In addition, pigment forming bacteria can contribute to skin UV
protection24. erefore, it was our hypothesis that pomegranate consumption might alter the skin microbiota
contributing to protection from UVB.
e objective of the current study was to determine whether pomegranate extract (PomX) or pomegranate
juice (PomJ) can decrease UVB-induced skin photoaging, alter inammatory markers and the skin microbiota.
Results
Characteristics of participants. Seventy-four participants completed the study. ere was no statistically
signicant dierence at baseline for average age, height, weight, BMI, race, ethnicity and skin type (Table1).
Although there were more Asian women in the PomJ group compared to PomX and placebo, there was no dier-
ence in skin type distribution among the three groups (Table1).
Urolithin formation. In the intestine pomegranate ellagitannins are broken down to ellagic acid (EA), which
can be absorbed and converted to methylellagic acid glucuronide (DMEAG). Urinary DMEAG can be used to
determine compliance. EA remaining in the intestine is further metabolized by bacteria to urolithin A (UA). Aer
absorption UA circulates in form of UA glucuronide (UAG). In the PomJ group, 16 participants (67%) showed
UAG in urine and 8 (33%) were UA non-producers, while in the PomX group 19 participants (83%) formed
UA and 4 (17%) were UA non-producers. In the PomX group two participants had neither UAG nor DMEAG
Pom Juice (n = 24) Pom extract (n = 25) Placebo (n = 25) P value
Age (years) 35.1 ± 4.3 35.9 ± 4.1 37.9 ± 4.2 0.063
Height (inches) 63.5 ± 2.8 63.9 ± 2.6 63.8 ± 2.5 0.843
Weight (lbs) 153 ± 30.8 158.7 ± 34.5 170.2 ± 40.7 0.253
BMI 26.6 ± 5.0 27.1 ± 5.1 29.9 ± 6.7 0.092
Female 24 (100) 25 (100) 25 (100)
Race:White 14 (58) 20 (80) 14 (56) 0.284
Black 01 (4) 1 (4)
Asian 9 (38) 3 (12) 3 (12)
Bi-racial 1 (4) 1 (4) 2 (8)
Ethnicity:Hispanic 9 (37) 16 (64) 10 (40) 0.119
Non-Hispanic 15 (63) 9 (36) 15 (60)
Skin Type 0.552
II 5 (20.8) 2 (8.0) 3 (12.0)
III 6 (25.0) 9 (36.0) 11 (44.0)
IV 13 (54.2) 14 (56.0) 11 (44.0)
Melanin index (RU) 264.5 ± 212.6 242.7 ± 91.1 202.2 ± 75.4 0.59
UAG producer 16 (67) 19 (83) n/a
UAG non-producer 8 (33) 4 (17)# n/a
Table 1. Baseline demographics of study participants (n = 24–25). Data are mean ± SD. Numbers in
parenthesis are percent. #two participants did not produce urolithin A glucuronide (UAG) or dimethylellagic
acid glucuronide (DMEAG), relative unit RU.
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in urine, which may indicate that on the day prior to the blood collection the participants did not consume the
pomegranate product. ey were excluded from the statistical analyses of the skin measurements. ere was no
signicant dierence in urine UAG and DMEAG between individuals consuming PomJ or PomX (Table2).
Eects of POM extract consumption on minimal erythema dose and melanin index. To e val -
uate the minimal erythema dose (MED) the inner arm skin was exposed to increasing dose and exposure time
to determine the minimal UVB dose that induces erythema. In women consuming PomJ (p = 0.038) or PomX
(p = 0.011) for 12 weeks the MED was increased signicantly when analyzing the dierence of baseline to nal
erythema dose compared to placebo group. In the PomX group UA producers experienced a signicant increase
in MED, while in UA non-producers MED was not increased signicantly (Table2). In the PomJ group, there
was no dierence between UA producers and non-producers in regards to MED. e time of exposure showed
a trend to increase in both groups compared to the placebo group (p = 0.083 for PomJ and p = 0.089 for PomX).
Upon exposure to UVB irradiation epidermal melanocytes produce melanin that is transferred to neighboring
keratinocytes protecting the cells from UV radiation damage25. However, hyperpigmentation will lead to prema-
ture photoaging and senescence in melanocytes25. Melanin formation also can be used to evaluate the eect of
UVB exposure. In the present study in women consuming PomJ and PomX melanin concentration was decreased
but did not reach signicance (Table3).
Eects of POM extract consumption on the skin microbiota. Pomegranate ellagitannins and EA
have anti-bacterial activity in vitro and topical application of a Pom extract ointment has been shown to decrease
the growth of Propionibacterium acnes and reduce edema in Wistar rat ears23,26,27. In the present study we deter-
mined whether oral consumption of PomJ or PomX altered the microbiota on the skin surface. Microbiota anal-
ysis showed the following composition of the skin microbiota on the phylum level: 36–38% Firmicutes, 25–31%
Proteobacteria, 18–30% Actinobateria, 9–17% Bacteroidetes, 0.2–1.9% Fusobacteria, 0.2–1.1% Cyanobacteria,
0.003–0.4% Tenericutes and 0.2–1% others (Fig.1A). Intergroup analysis showed no signicant dierence in
change of phylum composition of skin samples collected at baseline and aer 12 week intervention from women
consuming PomJ compared to placebo and PomX and placebo.
Intergroup analysis comparing the change in relative abundance of skin bacteria from baseline to week 12
between each intervention group (placebo, PomX and PomJ) showed multiple signicant changes, which were
not always in the same direction for PomX and PomJ intervention. We focused our data analysis on bacteria with
relative abundance that was signicantly dierent between the pomegranate and placebo groups.
Pom Juice UA
producer (n = 16) Pom Juice nonUA
producer (n = 8) Pom extract UA
producer (n = 19) Pom extract nonUA
producer (n = 4) Placebo
(n = 25)
Urine UAG (ng/mL) 3147 ± 1861 0 4068 ± 2049 0 n/a
Urine DMEAG (ng/mL) 163 ± 147 199 ± 118 210 ± 155 200 ± 101 n/a
MED BL (mJ/cm2) 385.7 ± 97.7 379.3 ± 100.1 409.2 ± 80.6 347.5 ± 111.4 384.2 ± 105.6
MED F (mJ/cm2) 417.5 ± 126.6 420.0 ± 87.6 440.0 ± 88.1*373.8 ± 41.1 367.2 ± 90.2
Table 2. Minimal erythema dose and urine urolithin A glucuronide and dimethylellagic acid glucuronide
in UA producers and non producers consuming PomJ, PomX or placebo. Data are mean ± SD, n = 24–25.
ANCOVA model was used to compare outcomes adjusted for baseline value between placebo and Pom extract
or placebo and Pom Juice groups; *p < 0.05. Urolithin A glucuronide (UAG); dimethylellagic acid glucuronide
(DMEAG); minimal erythema dose (MED); baseline BL; nal (F).
Pom Juice (n = 24) Pom extract (n = 25) Placebo (n = 25)
Compliance F (%) 96.3 ± 5.1 97.5 ± 3.9 98.9 ± 2.1
MED BL (mJ/cm2) 383.6 ± 95.8 396.8 ± 83.6 384.2 ± 105.6
MED F (mJ/cm2) 418.3 ± 113.1*429.6 ± 81.8*367.2 ± 90.2
Time BL (sec) 175.5 ± 42.9 193.7 ± 46.7 185.6 ± 52.5
Time F (sec) 199.1 ± 58.6 203.6 ± 49.0 177.2 ± 43.6
Melanin index BL (RU) 264.5 ± 212.6 242.7 ± 91.1 202.2 ± 75.4
Melanin index F (RU) 195.0 ± 63.9 219.6 ± 61.0 198.9 ± 67.4
Sebum BL (µg/cm2) 17.0 ± 19.7 14.8 ± 17.5 22.1 ± 36.8
Sebum F (µg/cm2) 15.3 ± 24.4 15.2 ± 18.2 26.9 ± 37.7
Hydration BL (RU) 41.8 ± 14.4 36.4 ± 12.6 41.9 ± 14.5
Hydration F (RU) 38.7 ± 9.8 41.4 ± 11.4 40.9 ± 10.8
Table 3. UVB-induced minimal erythema dose, time of exposure and skin characteristics determined before
and aer PomX, PomJ and placebo intervention. Data are mean ± SD, n = 24–25. BL = baseline, F = nal,
RU = relative unit. ANCOVA model was used to compare outcomes adjusted for baseline value. *p < 0.05.
Minimal erythema dose (MED); baseline BL; nal (F).
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On the family level bacteria from the family Aerococcaceae, Methylobacteriaceae and Campylobacteraceae
were altered signicantly with PomX consumption compared to placebo (Fig.2). Intergroup analysis comparing
the change of bacterial abundance from week 12 to baseline showed that Campylobacteraceae (Proteobacteria)
was increased and Methylobacteriaceae not changed in the PomX group, while a decrease was observed in the
placebo group (Fig.2). e relative abundance of Aerococcaceae was increased in the placebo group, but there
was no change in the PomX group (Fig.2). e changes for PomJ compared to placebo group were not sig-
nicant. On the genus level the most frequently occurring bacteria included Propionibacterium (Actinobacte
ria) > Staphylococcus (Firmicutes) > Prevotella (Bacteroidetes) > Streptococcus (Firmicutes) > Lactobacillus
(Firmicutes) > Corynebacterium (Actinobacter) > Veillonella (Firmicutes) > Haemophilus (Proteobacteria) > A
cinetobacter (Proteobacteria) (Fig.1B). On the genus level comparing the PomX to placebo group, ve genera
were changed in opposite direction compared to the placebo group: Coprococcus (Firmicutes) was decreased,
Alicycliphilus, Conchiformibius and Campylobacter (Proteobacteria) were increased and Geodermatophilaceae_
unclassied was not changed comparing week 12 to baseline (Fig.3). Comparing PomJ group to placebo we
determined that the unclassied genus from the family Rhizobiaceae was changed signicantly (Fig.3).
Pomegranate consumption had no eect on alpha diversity and no clusters were observed in the principal
coordinate analysis of weighted and unweighted beta-diversity (Figs4 and 5).
Discussion
e main ndings observed in the present study were a signicant increase in minimal UVB dose to induce ery-
thema in women consuming 1000 mg of PomX or 8 oz of PomJ daily for 12 weeks. e minimal erythema dose is
determined by the lowest UVB dose and time of exposure to induce skin erythema. In the present study we also
observed a non-signicant trend to increase in time of UVB exposure (Pom X p = 0.08 and PomJ p = 0.088) and
a non-signicant decrease in melanin formation (Pom J p = 0.16). Together the results demonstrate that pome-
granate consumption may lead to increased protection to UVB-induced damage to skin.
Possibly the MED might have been increased due to systemic pomegranate metabolites such as dimethylel-
lagic acid glucuronide or urolithin A glucuronide circulating in the blood stream. Since we found that in the Pom
X group urolithin A producers showed a signicant increase in MED compared to non-producers, we suggest that
circulating urolithin A glucuronide is involved in the UVB protection.
In previous in vitro and animal studies the topical application of pomegranate and EA improved the resist-
ance of skin to UVB exposure12. For example, the study by Bae JY et al. demonstrated that topical application of
EA reduced collagen breakdown by inhibiting matrix metalloproteinase (MMP) activity and inammation in
UVB-irradiated human skin cells and hairless mice12. In addition, animal studies also showed that oral consump-
tion of pomegranate and EA prevented UVB-induced skin damage4,28,29. For example oral consumption of PomJ
concentrated powder in hairless mice resulted in reduction of UVB-induced skin wrinkles through increased skin
water content, collagen type I and hyaluronan content4. Other processes reported in the literature, contributing to
pomegranate’s UVB photoprotection of the skin include oxidation, inammation, melanin formation, apoptosis
Figure 1. Relative abundance of skin microbiota before and aer pomegranate and placebo intervention.
Stacked column bar graphs depict the average relative abundance and distribution of the most abundant
resolved taxa at the phylum (A) and genus (B) level before (BL) and aer (12 week, F) PomX, PomJ and placebo
control (Cntr) intervention.
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Figure 2. Relative abundance of skin bacteria at the family level that were signicantly dierent comparing
PomX to placebo control (Cntr) and PomJ to placebo control groups (change from BL to 12 weeks [F]). Data
are mean ± SD, n = 24–25. Non-parametric Kruskal–Wallis with Mann-Whitney test was used. Bonferroni
correction was used to correct the probability for multiple comparisons.
Figure 3. Relative abundance of skin bacteria at the genus level that were signicantly dierent comparing
PomX to placebo control (Cntr) and PomJ to placebo control groups (change from BL to 12 weeks [F]). Data
are mean ± SD, n = 24–25. Non-parametric Kruskal–Wallis with Mann-Whitney test was used. Bonferroni
correction was used to correct the probability for multiple comparisons.
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of keratinocytes, activity of matrix metalloproteinases (MMPs), collagen and elastin formation and were identi-
ed in in vitro studies in human skin broblasts, keratinocytes or reconstituted skin13,14,30.
Previously, one human study has been performed testing the eect of oral consumption of a high EA pome-
granate extract for 4 weeks on pigmentation in the skin caused by UV irradiation and showed a trend, but no
signicant change in erythema17. Possibly, in this study by Kasai et al. 4 weeks of pomegranate consumption was
not long enough to see a signicant change. Another human study demonstrated that oral consumption of a phy-
tonutrient blend containing omega-3 fatty acids, resveratrol, quercetin, and other polyphenols led to protection
against UVR-induced skin damage31.
Skin exposed to UVB irradiation develops symptoms of a mild sunburn associated with inflammatory
response and characterized clinically by redness, which is mediated by increased dermal vascular permeability,
vasodilation, edema and inammatory cell inltration32. In the current study we did not observe a change in skin
hydration at the end of pomegranate consumption for 12 weeks.
A likely target of Pom metabolites is melanin formation28,30. We observed a trend to decrease of melanin
formation comparing melanin concentration before and aer pomegranate consumption. It has been previously
demonstrated that EA and pomegranate concentrate inhibited tyrosinase activity, the enzyme necessary for mel-
anin formation28. Administration of pomegranate extract inhibited pigmentation, in a dose-dependent manner,
Figure 4. Diversity analyses of skin microbiota before (BL) and aer (F) PomX, PomJ and placebo control
(Cntr) intervention. Alpha diversity using Chao1 index (A) and Whole tree index (B) was evaluated using
QIIME soware package. Data are means ± SD (n = 24–25).
Figure 5. Beta-diversity analyses of skin microbiota before (BL) and aer (F) PomX, PomJ and placebo control
(Cntr) intervention. Unweighted (A) and weighted (B) UniFrac PCoA plots were created using QIIME soware
package.
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on the skin of brownish guinea pigs receiving UV irradiation, where the number of melanocytes in the epidermis
was decreased in a dose-dependent manner28. ese results suggest that EA orally administered is absorbed into
the body and the EA and/or its metabolites inhibit proliferation of melanocytes in the skin, resulting in inhibited
synthesis of melanin by tyrosinase in melanocytes.
Other potential mechanisms of protection from photoaging are through anti-apoptotic eects, inhibition of
MMP activity, inhibition and extracellular matrix (ECM) (COL1 and hyaluronan) synthesis- related moistur-
izing, anti-inammatory and antioxidative eects, which have been observed in healthy female SKH-1 hairless
mice receiving oral gavage of 100–400 mg/kg body weight of a dried PomJ concentrated powder4. In the present
study, we did not nd any changes in gene expression of markers of inammation (Smad3, Tgfβ) and photoaging
(MMP1, β-integrin, stratin and IGF1R) (data not included). Samples were collected by tape stripping of skin
locations not exposed to UVB since UVB exposed skin was irritated and tape stripping could cause considerable
skin irritations. However, since no photo challenge was induced to the skin prior to tape stripping we did not
observe any protection with pomegranate consumption.
Human skin microbiota composition depends on the location of the skin on the body33. Skin swabbing sam-
ples for microbiota analysis were collected at the inner elbow (antecubital fossa), which is considered a moist but
not oily habitat. is region is usually enriched for Corynebacteria species and Staphylococci species34. In healthy
individuals, the most common skin bacteria are categorized into four dierent phyla: Actinobacteria (most dom-
inated by Propionibacterium spp., and Corynebacterium spp.), Firmicutes (major genus is Staphyloccocus spp.),
Proteobacteria and Bacteroidetes33,35. Previous studies have demonstrated that consumption of pomegranate will
aect the intestinal microbiota36 and in vitro studies demonstrated the antibacterial eect of pomegranate on
bacteria commonly found on skin, such as Propionibacterium and Staphylococcus23. However, the present study
is rst to demonstrate that oral consumption of pomegranate altered the skin microbiota. Skin swipes for the
microbiota assessment were performed in skin areas of the inner arm, which were not exposed to photoaging. No
changes on the phylum level were observed. On the family and genus level several bacteria with minor abundance
were changed when comparing PomX and PomJ to placebo. Bacteria in the family Methylobacteriaceae (phy-
lum Proteobacteria) have been found to form UVA-absorbing compounds and are frequently found on plant’s
phyllosphere (above ground), exposed to harmful UV irradiation24. e proportion of Methylobacteriaceae was
decreased in the placebo and PomJ groups and not changed in the PomX group comparing 12 weeks to base-
line. e abundance of Methylobacteriaceae was very low and little is known about pigments in other bacteria.
Bacterial pigments, however, can potentially contribute to UV skin protection. Other bacteria with altered abun-
dance such as Campylobacter (Proteobacteria) and Coprococcus (Firmicutes) are not commonly found on skin.
Our intergroup evaluation of bacteria abundance showed dierent eects of PomX and PomJ consumption on the
skin microbiota. However, both extract and juice intervention resulted in an increase in UVB protection. Possibly
the skin microbiota does not contribute to the UVB protection or since the microbiota functions in a network of
many bacteria possibly changes in individual bacteria may not aect the microbiota function.
The evaluation of the skin microbiota was performed in healthy skin at a site without UVB exposure.
erefore, we do not know if the observed changes in the skin microbiota contributed to the increase in UVB
protection.
One shortcoming of this study was that it is not possible to sample skin sites shortly aer UV exposure. In future
studies we will collect skin samples near the site of UV exposure to evaluate the mechanism of skin UVB protection
from UVB-induced photodamage and to determine if the skin microbiota contributes to UVB protection.
In summary, PomX and PomJ consumption resulted in an increase in skin protection to UVB exposure
as shown by an increase in minimal erythema dose and a trend to decrease melanin formation, indicating an
enhancement of UVB protection. Our mechanistic studies did not provide insights into specic targets of the
pomegranate induced skin protection mostly because skin and microbiota samples were collected at non-UVB
exposed sites. In future studies, we will collect samples near exposed sites to test if changes in gene expression and
composition of the microbiota contribute to the UVB protection by pomegranate consumption.
Methods
is study was a randomized controlled, parallel, three arm open label study completed at the Center for Human
Nutrition, University of California Los Angeles, California, USA. e study was carried out in accordance with
the guidelines of the Human Subjects Protection Committee of the University of California, Los Angeles. e
clinical protocol was approved by the internal review board (IRB) of the University of California, Los Angeles.
All subjects gave written informed consent before the study began. e study was registered in ClinicalTrials.gov
under the following identier: NCT02258776 on 10/07/2014.
Study participants. Seventy-seven healthy women were enrolled. Seventy-four women completed the
12-week pomegranate intervention study. ree participants dropped out related to pregnancy (one participant
in the PomX group and one in the PomJ group) and related to moving out of state (one particiant in the the PomJ
group). eir data was not included in the statistical evaluation. No adverse eects were reported. Inclusion criteria
were 30–40 years of age, female, be in good health. Exclusion criteria were: no use of topical antibiotic or topical
steroid on the face, scalp, neck, arms, forearms or hands in the previous 7 days or with any skin condition in the
target area, no skin irritations, dry skin or rash and no intake of antibiotics. roughout the study participants were
instructed not to consume pomegranate products, walnuts, or polyphenol-rich fruits (strawberry, raspberry, etc.).
Study design. Participants were randomized to consuming either pomegranate juice (PomJ 8 oz)(n = 24),
pomegranate extract capsules (PomX 1000 mg) (n = 25), or placebo (n = 25) capsules for 12 weeks. e random
permuted block design was implemented to carry out the randomization using our standard random number
program. Block size was 4 or 6 carried out in a random way. Subjects were instructed to take a daily dose of
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1000 mg of the pomegranate extract (POMx®, POM Wonderful, Inc., Los Angeles), which delivers pomegranate
polyphenols in an amount equivalent to about 8 oz of pomegranate juice. PomX was developed to be used as a
dietary supplement and has Generally Recognized as Safe status37. Pomegranate placebo capsules contained inac-
tive excipients (dextran). PomJ contained 100 mg of punicalagin A/B and 23 mg ellagic acid in 240 ml of juice, and
PomX contained 100 mg punicalagin A/B and 44 mg ellagic acid in 1000 mg extract, as determined by HPLC38.
Aer subjects signed the informed consent form at baseline, weight and height were measured and skin type
was evaluated. At baseline prior to taking pomegranate and at the end of 12 weeks of pomegranate consumption,
evaluation of UVB-induced erythema, microbiota, gene expression of epithelial cells and blood collection was
performed as described below.
Study outcomes. e primary outcome of this intervention study was the quantication of the minimal
erythema dose (MED) of skin response to UVB exposure and the measure of melanin index in skin. For the
secondary outcome changes in the skin microbiota were determined. Both outcomes were determined before
(baseline) and aer (12 weeks) of PomJ, PomX or placebo consumption.
Minimal erythema dose, melanin index, hydration and sebum evaluation. We determined
the lowest dose of UVB radiation capable of inducing erythema (minimal erythema dose [MED]). e MED
was determined for each subject before (week 0) and aer (week 12) the intervention. Prior to testing, the skin
type was evaluated based on the Fitzpatrick Skin Type scale39. Participants with Fitzpatrick skin type 2–4 were
included in the study. Background erythema (T0) was measured in all test areas before treatment using a mex-
ameter probe attached to Cutometer dual MPA 580 (Courage&Khazaka electronic GmbH, Koeln, Germany).
Skin UVB dose and treatment time were determined based on overall skin type classication. Using the dosing
guideline for NB-UVB and the National Biological UVB mJ chart, we determined the sequential exposure times
for each skin patch. A sleeve with 6 cut out patches was placed on the subjects arm. Using the Dermalight 90
handheld device (National Biological, Beachwood, OH) the test area on the inner arm of subjects was irradiated
with a dened dose of narrow band ultraviolet B (NB-UVB) light delivered by the UV radiation between 270 and
400 nm, peaking at 310 nm was delivered from a uorescent UV-B lamp (Philips TL20 W/12). Depending on skin
type, a dose range of 220–550 mJ/cm2 for a time of 100–290 seconds was used. is dose range is typically used
for treatment of skin conditions like psoriasis and vitiligo. To evaluate MED the subject returned 24 hours later
to determine which skin patch showed minimal erythema (pink color). Photographs were taken before irradia-
tion and aer 24 hours. e lowest dose and time of the occurrence of pink were determined and used as MED.
Melanin index, hydration and sebum on skin surface were evaluated at baseline (prior to Pom intake) and aer
12 week intervention using the mexameter MA18, corneometer CM825 and sebumeter SM815 probes attached
to the Cutometer. Sebumeter SM 815 uses the dierence of light intensity through a plastic strip to indicate the
amount of absorbed sebum. e sebum level is expressed in μg/cm2 40. Corneometer CM 825 uses the high dielec-
tric constant of water for analyzing the water-related changes in the electrical capacitance of the skin. It displays
hydration measurements in system-specic arbitrary units40. A melanin index is calculated by Mexameter MX 18
from the strength of the absorbed and the reected light at, respectively, 660 and 880 nm. An erythema index is
processed similarly at, respectively, 568 and 660 nm40.
Skin surface microbiota collection. Skin sampling using the wet swab method was performed at base-
line and 12 weeks. Samples were collected as described by the Human Microbiome Project41. A sterile 4 cm
square template was placed on the inner arm to mark the sampling area. e collection swab (CatchAll®Sample
Collection Swab (Epicenter, Illumina, Madison, WI) was moistened with buer (50 mM Tris buer [pH 7.6],
1 mM EDTA [pH 8.0], and 0.5% Tween-20) and the area within the template was swabbed for 30 s rubbing the
swab back and forth about 50 times applying rm pressure. e swabs were placed into bead solution for DNA
extraction using DNeasy Powerlyzer microbial kit (Qiagen, Valencia, CA) and vortexed for 30 sec. e quality of
the extracted DNA was conrmed using the Nanodrop 1000 (ermo Fisher Scientic, Wilmington, DE).
Skin microbiological analyses. MiSeq sequencing. Microbial sequencing of the V1 to V3 region of 16 S
bacterial rDNA was performed using primer pair 27 F (AGA GTT TGA TCC TGG CTC AG) and 534 R (ATT
ACC GCG GCT GCT GG)42. 30 cycle PCR using the HotStarTaq Plus Master Mix Kit (Qiagen, USA) was per-
formed under the following conditions: 94 °C for 3 min, followed by 28 cycles of 94 °C for 30 s, 53 °C for 40 s and
72 °C for 1 min, aer which a nal elongation step at 72 °C for 5 min was performed. Aer amplication, PCR
products were checked in 2% agarose gel to determine the success of amplication and the relative intensity of
bands. Multiple samples are pooled together (e.g., 100 samples) in equal proportions based on their molecular
weight and DNA concentrations. Pooled samples were puried using calibrated Ampure XP beads. en the
pooled and puried PCR product was used to prepare DNA library by following Illumina TruSeq DNA library
preparation protocol. Sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA)
on a MiSeq (Illumina, San Diego, CA) following the manufacturer’s guidelines. Sequence data were processed
using a proprietary analysis pipeline (MR DNA, Shallowater, TX, USA). In summary, sequences were depleted
of barcodes then sequences <150 bp removed, sequences with ambiguous base calls removed. Sequences were
denoised, OTUs generated and chimeras removed. Operational taxonomic units (OTUs) were dened by cluster-
ing at 3% divergence (97% similarity). Final OTUs were taxonomically classied using BLASTn against a curated
GreenGenes database43. Within community diversity (α-diversity) was calculated using Quantitative Insights
Into Microbial Ecology (QIIME) soware package44. Analysis of α-diversity (Shannon index) was performed by a
one-way ANOVA. β-diversity was measured by calculating the weighted UniFrac distances45 using QIIME default
scripts, and weighted UniFrac PCoA biplot was visualized using EMPeror46.
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Identication of pomegranate metabolites by high performance liquid chromatography and
mass spectrometry. All solvents were HPLC grade from Fisher Scientic. Ellagic, formic and phosphoric
acid were purchased from Sigma-Aldrich (St Louis, MO). Pure punicalagin A/B was purchased from ChromaDex
(Irvine, CA) and urolithin A was purchased from Jinan Feiteng Technology (Jinan Shandon, China). e com-
position of the pomegranate extract was analyzed by HPLC and diode array detection. To determine the concen-
tration of urolithin A glucuronide and dimethylellagic acid glucuronide in urine, samples (1 mL) were diluted
with 1 ml of 2% formic acid MeOH, vortexed for 30 s and centrifuged at 20,000xg for 10 min at 4 °C. e super-
natant was analyzed by LC-MS/MS8. e concentration was estimated based on urolithin A standard. e con-
version of urolithin A glucuronide to urolithin was estimated by using β-glucuronidase to catalyze hydrolysis of
β-D-glucuronic acid residues from urolithin A glucuronide in human urine samples.
Statistical analysis. To obtain an estimate of the power of this study to detect a treatment eect for the pri-
mary outcome we use data from Kasai et al.17. On the basis of this data we estimated power for this study assum-
ing a treatment eect of similar magnitude and we use a two sample t-test. Based on these assumptions, a nal
sample size of 20 per group will have 85% power to detect a dierence in mean over time among the groups with a
0.050 two sided signicance level. We assume that there will be a 20% drop out rate and the goal was to randomize
24 subjects per group. e study was stopped when at least 24 subjects for each group completed the intervention.
Summary statistics (mean, standard deviation and frequency distribution) were generated for baseline demo-
graphic and clinical information for each study group to characterize the study population. ANOVA (analysis
of variance) and Chi-square test were used to evaluate the dierence between treatment groups for continuous
variable and categorical variables, respectively. e 12 week outcomes were compared between study groups,
using ANCOVA (analysis of covariance) with the adjustment of baseline values. Data management, variable
transformations, and other statistical analyses were conducted using SAS 9.2 (Statistical Analysis System, Cary,
NC, 2008). Dierence in changes of bacterial relative abundances over 12 week intervention among the treatment
groups were compared using the non-parametric Kruskal–Wallis test in IBM SPSS Statistics version 23. Only
the signicant genera and species from the Kruskal–Wallis test were further tested with Mann–Whitney test to
assess the dierences between treatments. Bonferroni correction was used to correct the probability for multiple
comparisons. P values < 0.05 were considered statistically signicant.
Data Availability
e datasets generated during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Supported by departmental funds from the Center for Human Nutrition, Department of Medicine, David Geen
School of Medicine, University of California, Los Angeles.
Author Contributions
S.M.H. wrote the main manuscript text, J.Y., Y.X., J.L., E.H.P. and M.H. performed QIIME and microbiota data
analysis, R.P.L. and J.H. performed chemical analyses, I.G. and G.T. coordinated clinical study, J.K., Z.L. and
D.H. participated in manuscript preparation and C.H.T. performed statistical analysis. All authors reviewed the
manuscript.
Additional Information
Competing Interests: e authors declare no competing interests.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and
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... Results demonstrated improved resistance to UVB exposure after 8 weeks of consumption of pomegranate extract and juice, but no effect on skin elasticity and firmness. 16 We have found similar results with daily consumption of 1.5 oz of almonds daily. 17 It was the objective of the present study to investigate the effects of consumption of one avocado daily compared with no avocado control on skin health including elasticity, firmness, hydration, pigmentation, and UVB resistance in women with increased abdominal circumference and elevated BMI. ...
... We determined the lowest dose of UVB radiation capable of inducing erythema (MED) as described in detail in our previous publication. 16 ...
... The resistance of the skin to negative pressure (ie, firmness) and its ability to return to its original position (ie, elasticity) are re- Melanin content and erythema were measured using a mexameter MA18 probe, hydration using the corneometer CM825 probe, and sebum using the sebumeter SM815 probe attached to the Cutometer dual MPA 580 (Courage&Khazaka electronic GmbH, Koeln, Germany) as previously described. 16 The measurements were performed in triplicates at two locations (forehead and 1 inch below the outer edge of the eye). The software of the cutometer MPA 580 allows the analysis of several parameters of skin elasticity and firmness. ...
Article
Background Avocados are a rich dietary source of monounsaturated fatty acids, carotenoids, and phenolic compounds. Clinical studies have demonstrated that oral consumption of carotenoids improved skin aging. However, no studies have investigated whether oral intake of avocado will reduce skin aging. Objectives We therefore performed this pilot study to assess whether oral consumption of one avocado daily for 8 weeks can reduce skin aging in healthy overweight women assessing skin physical characteristics and resistance to UVB radiation. Methods Thirty-nine female participants (age 27–73 years) with Fitzpatrick skin type II-IV were randomly assigned to consume either one avocado daily or continue habitual diet for 8 weeks. Facial skin elasticity, firmness, pigmentation, sebum, and hydration were determined using a cutometer on the forehead and under eye. Minimal erythema dose (MED) was determined by standardized protocol at inner arm. Results Elasticity and firmness were increased at forehead comparing 8 weeks to baseline in the avocado group. Comparing avocado to control, change in firmness marker from baseline to week 8 indicated a significant increase in forehead skin firmness in the avocado group. We did not observe any change in hydration, pigmentation, sebum, and UVB resistance between the avocado and control group, although changes in melanin and erythema were observed in both groups over time. Conclusions Our findings suggest that daily oral avocado consumption may lead to enhanced elasticity and firmness of the facial skin in healthy women. Further studies of other skin locations are required to establish the connection between avocado consumption and skin aging.
... Henning et al. [283] randomized 74 women to receive 8 oz of PJ (23 mg EA), 1000 mg of pomegranate extract (same dose of EA as juice), or placebo for twelve weeks. They found that either the liquid or the placebo significantly increased the minimal erythema dose. ...
... They found that either the liquid or the placebo significantly increased the minimal erythema dose. Therefore, it may lead to increased UV damage protection [283]. ...
... After one year of treatment, there was a statistically significant decrease in CAIMT (p < 0:01). The authors do not specify Induce improvement in patients with hyperpigmentation and dark spots [280] Patients with melasma showed significant improvement after treatment [281,282] Lead to increased UV damage protection [283] Have antiwrinkle activity, increasing the expression of type I collagen mRNA [284] Insulin resistance and diabetes Induce reduction in blood glucose levels [285,286] Weight and body composition No significant effect on body weight, body mass index, waist circumference, or body fat percentage [287] Cardiovascular disease risk factors Decrease carotid artery intima-media thickness; decrease in mean peak systolic velocity and end-diastolic velocity of carotid arteries [288] Reduction in stress-induced ischemia [289] Significant reductions in systolic and diastolic blood pressure [290,291] No effect on plasma concentrations of cholesterol, LDL-C, HDL-C, or triglycerides [292,293] ...
Article
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Ellagic acid (EA) is a bioactive polyphenolic compound naturally occurring as secondary metabolite in many plant taxa. EA content is considerable in pomegranate (Punica granatum L.) and in wood and bark of some tree species. Structurally, EA is a dilactone of hexahydroxydiphenic acid (HHDP), a dimeric gallic acid derivative, produced mainly by hydrolysis of ellagitannins, a widely distributed group of secondary metabolites. EA is attracting attention due to its antioxidant, anti-inflammatory, antimutagenic, and antiproliferative properties. EA displayed pharmacological effects in various in vitro and in vivo model systems. Furthermore, EA has also been well documented for its antiallergic, antiatherosclerotic, cardioprotective, hepatoprotective, nephroprotective, and neuroprotective properties. This review reports on the health-promoting effects of EA, along with possible mechanisms of its action in maintaining the health status, by summarizing the literature related to the therapeutic potential of this polyphenolic in the treatment of several human diseases.
... It confers photoprotection through inhibition of UV-induced production of free radicals, erythema and burning, DNA damage, cell proliferation, and apoptosis [8,24] . It can also decrease collagen breakdown [25] . ...
... Oral administration of pomegranate extract was found to be protective against UVB-induced skin cancer formation in mice through downregulation of COX-2, iNOS, cyclin D2, and MMPs. Women who consumed 1000 mg of pomegranate extract or 8 ounces of pomegranate juice daily for 12 weeks were reported to have increased MED in a randomized controlled, open-label study [25] . ...
Article
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The importance of photoprotection against the deleterious effects of excessive and chronic exposure to sunlight is now well established. Photoprotective measures include behavioral modifications such as seeking shade, wearing photoprotective clothing, wide-brimmed hat and sunglasses, and applying sunscreen to exposed areas. Data on botanical topical and oral preparations have demonstrated photoprotective potential in in vitro, animal, and human studies. This review will focus on botanicals that have been most extensively studied, namely, Polypodium leucotomos extract, green tea, pomegranate, resveratrol, curcumin, and silymarin. These agents have shown promise in mitigating ultraviolet-induced acute changes on the skin, chronic photodamage, and even skin cancer prevention. However, it must be emphasized that current evidence indicates that these agents should be used as adjunctive measures rather than as a replacement of the photoprotective behavioral modifications described above.
... The extract and juice of P. granatum are known for their concentration of hydrolysable tannins (ellagitannins), including ellagic acid, as well as anthocyanins and other polyphenols [74]. At the end of that study, the hypolipidemic effect was observed only in individuals with metabolite B, along with reductions in total cholesterol, LDLcholesterol, non-apolipoprotein-B cholesterol and LDLcholesterol [72,75]. ...
... In an obesogenic rat model, the proanthocyanidin of grape seed extract increased adipocyte hyperplasia and reduced the hypertrophy of the adipose retroperitoneal adipose tissue through inhibition of adipogenesis regulation by PPARγin a sirtuin 1 (Sirt1)-dependent manner. Hyperplasia is associated with the prevention of obesity-related metabolic disorders [75]. In cardiovascular diseases, proanthocyanidins exert their protective effect by inhibiting the oxidation of LDL-cholesteroland binding this molecule to the lectin-like oxidized LDL-1 receptor (LOX-1) involved in the pathogenesis of arteriosclerosis. ...
Article
Full-text available
The fruit and leaves of Eugenia dysenterica DC., locally known as cagaita, are rich in antioxidant glycosylated quercetin derivatives and phenolic compounds that have beneficial effects on diabetes mellitus, hypertension and general inflammation. We conducted a literature search to investigate the nutraceutical potentials of these phenolic compounds for treating obesity, diabetes mellitus and intestinal inflammatory disease. The phenolic compounds in E. dysenterica have demonstrated effects on carbohydrate metabolism, which can prevent the development of these chronic diseases and reduce LDL (low-density lipoprotein) cholesterol and hypertension. E. dysenterica also improves intestinal motility and microbiota and protects gastric mucosa, thereby preventing inflammation. However, studies are necessary to identify the mechanism by which E. dysenterica nutraceutical compounds act on such pathological processes to support future research.
... This plant is found in tropical and subtropical regions and is widely used for hypertension and atherosclerosis. A few in vitro and in vivo animal studies have already been carried out to evaluate the cutaneous impact of this extract, mainly in inhibiting melanin synthesis, improving hydration and elasticity and preventing skin ageing, in addition to antioxidant and anti-inflammatory activity and protection against damage from the UV exposure [1,[77][78][79][80][81][82][83]. ...
... Henning et al. [77] evaluated in vivo the performance of oral supplementation of juice and capsules containing pomegranate extract against premature aging caused by UV radiation. This trial was carried out for 12 weeks on female volunteers aged between 30 to 40 years old and with skin phototype II to IV, which were separated into three distinct groups: placebo, those who consumed pomegranate juice and those who received supplementation with capsules containing 1000 mg of pomegranate extract. ...
Article
Full-text available
Skin health is not only significantly affected by ageing, but also by other lifestyle-related factors, such as sun exposure, exercise and eating habits, smoking or alcohol intake. It is known that the cutaneous tissue can exhibit visible signs of senescence, in the form of, for example, dull complexion, loss of firmness, or changes in pigmentation. Consumers attempt to improve skin health and appearance not only by cosmetic products, but also with the consumption of food supplements. Recently, there has been an increase in the amount of food supplements with claims that are related to skin and hair health. Nevertheless, the literature is still scarce in evidence of the efficacy of this type of products. Considering this scenario, we aim in this review to assemble studies and methodologies that are directed at the substantiation of the cutaneous health claims of food supplements. For example, we reviewed those that were indicative of antioxidant properties, improvement in pigmentation disorders, increased hydration or protection against the damages caused by ultraviolet radiation.
... The prevalence of pigmentation secondary to Minocycline ingestion varies between 2.4% to 41% of treated individuals and is highest in patients with rheumatoid arthritis 15 . Pomegranate fruit have been used for centuries in ancient cultures for its medicinal purposes and its benefits has been attributed to the content of hydrolysable tannins (ellagitannins) including ellagic acid and ellagic acid derivatives 16 . Ellagic acid is present in the extract of pomegranate has been proven to exhibit the potent antioxidant and tyrosine inhibitor properties 17 . ...
Article
Full-text available
Objective: This study was planned over the hypothesis that pomegranate extract rich in ellagic acid used with minocycline could decrease its adverse effect and prolong its therapeutic use and efficacy. Study design and Setting: This experimental study was done in the department of anatomy, Basic Medical Sciences Institute, Jinnah Postgraduate Medical Center, Karachi, Methodology: We acquired 40 guinea pigs (male, adult, 450 – 550 gm), randomly divided them into 4 groups.Group B received 0.0003mg/g bodyweight of minocycline only, group C was given 0.0003mg/g bodyweight of minocycline with 0.4mg/g bodyweight of pomegranate, group D was given 0.4mg/g bodyweight pomegranate only; with keeping group A with no intervention at laboratory diet for 8 weeks. After the experimental period, the animals were sacrificed, H & E and DOPA-OXIDASE staining was done on harvested skin tissues for morphometric observations under light microscopy. Results: The results showed that minocycline induced reduction in mean thickness of epidermis and increased melanin pigment deposition. Mean number of melanocytes decreased with pomegranate use though the difference was insignificant (P-value > 0.05) but consistent and measurable. Conclusions: It was proven that by including pomegranate in our daily diet, the process of hyperpigmentation of skin induced by the broad spectrum tetracycline particularly minocycline, can be slow down by decreasing the activity of tyrosinase enzyme, thus it provides a novel pathway to fight against any other drug induced hyperpigmentation occurring due to increase activity of tyrosinase enzyme
Article
Ethnopharmacological relevance Skin diseases are among the most common human health affections. A healthy skin promotes a healthy body that can be achieved through modern, allopathic and natural medicines. Therefore, medicinal plants can be a reliable therapy in treating skin diseases in humans through a diverse range of bioactive molecules they contain. Aim of the study This review aims to provide for the first-time scientific evidence related to the dermatological properties of Morocco's medicinal plants and it aims to provide a baseline for the discovery of new drugs having activities against skin issues. Methods This review involved an investigation with different search engines for Moroccan ethnobotanical surveys published between 1991 and 2021. The plants used to treat skin diseases have been determined. Information regarding pharmacological effects, phytochemical, and clinical trials related to the plants listed in this review was collected from different scientific databases like PubMed, Science Direct, Google Scholar, Web of Science and Scopus. The data were analyzed and summarized in the review. Results A total of 401 plants belonging to 86 families mainly represented by Asteraceae, Lamiaceae, Fabaceae, and Apiaceae which have been documented to be in common use by Moroccans for managing skin diseases. Among those plants recorded, the most commonly used are Allium cepa L, Chamaeleon gummifer (L.) Cass and Salvia rosmarinus Schleid. Mill. Leaves were the most commonly used plant part, while powder and decoction were the most common method of traditional drug preparation. 107 of the 401 plants (27%) have undergone pharmacological validation. A total of 44 compounds isolated from 27 plants were investigated to treat different types of skin diseases, and 25 plants have been clinically studied for their activities against skin diseases. Conclusion The beneficial effects of using Moroccan medicinal plants to treat skin diseases, according to traditional practices, have been proven in numerous scientific studies. Therefore, other studies should focus on isolating and identifying specific bioactive compounds from plant extracts, revealing more valuable therapeutic properties. Furthermore, additional reliable clinical trials are needed to confirm their beneficial effect on patients with skin diseases.
Article
Nuclear factor-kappa B (NF-κB) has important but complex functions in the photoaging of the human skin. This protein complex is activated upon UV irradiation and plays a key role in the signalling pathway of the inflammatory cascade. NF-κB induces the expression of various proinflammatory cytokines, such as tumour necrosis factor (TNF) and interleukin-1 (IL-1). These proinflammatory cytokines can in turn stimulate the activation of NF-κB, forming a vicious cycle. These processes cause chronic inflammation and contribute to skin ageing. In addition, the activation of NF-κB upregulates the expression of matrix metalloproteinases (MMPs) and leads to the degradation of structural proteins in the dermis. NF-κB disrupts the barrier function of the skin under prolonged and repeated UV stimulations in these ways. Such activity causes chronic skin damage, followed by the formation of wrinkles, dryness, roughness, laxity, and other photoaging manifestations. This study on the NF-κB signalling pathway and effector molecules provides a new perspective to understand and prevent photoaging.
Article
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Punica granatum L. belongs to the Punicaceae family which is distributed around the world. Different parts of pomegranate like seed, peel, juice, and leaves are rich in potential bioactive compounds. These plants have found application in traditional medicine such as in treatment of gastrointestinal, cardiovascular, and endocrine diseases, among others. The present review aimed to summarize the current research on the traditional and scientific applications of P. granatum with regard to the phytochemical content and clinical applications that may be useful for future drug development. Information about P. granatum was obtained from local classic herbal literature and electronic databases, such as PubMed, Scopus, and ScienceDirect. Several phytochemical constituents including polyphenolics, flavonoids, anthocyanosides, alkaloids, lignans, and triterpenes have been reported from the plant. Randomized clinical trials have provided evidence as to the pharmacological activities of pomegranate in several diseases including diabetes, cardiovascular disease, oral cavity disorders, endocrine disorders, and cancer. The present review has provided an insight into the traditional applications of the plants, and some of them have been validated by scientific evidence, particularly their applications as treatment of cardiovascular and endocrine diseases.
Article
The potential role of plant-based foods in the promotion of skin health is an emerging area of nutrition research. Plant-based foods are rich in bioactive compounds, including vitamin C, vitamin E, beta-carotene, polyphenols, and phenolic acids, which can contribute to oxidant defense, lower inflammation, and promote structural support of the skin. Epidemiological studies have associated higher intakes of select fruits and vegetables with positive skin health.1,2 Beneficial effects of certain fruits, vegetables, nuts, legumes, and polyphenolic-rich beverages on the skin have been reported, with each of these providing a unique phytochemical composition. While most studies use extracts, this review will focus on data from whole foods and minimally processed products. Collectively, the evidence to date suggests a promising future for plant-based dietary interventions that promote skin barrier health and function. However, additional research is required to address issues such as the optimal quality and duration of intake as well as potential mechanisms. Studies in the above areas will help formulate specific targeted dietary recommendations.
Article
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Skin is the largest body organ and the first barrier to exogenous threats. This organ is constantly exposed to external factors such as ultraviolet radiation, which induces many adverse effects including sunburn, depigmentation, photo aging, photo immune suppression, and even skin cancer. Antioxidants seem to be good candidates in order to reduce ultraviolet-mediated damages and to prevent the health consequences of ultraviolet exposure. The present investigation aims to further characterize the potential skin photoprotective effects of a food supplementation and a topical administration of a melon concentrate alone or in combination. A clinical study assessing the Minimal Erythema Dose (MED) was first set up to evaluate photoprotection. Afterward, an independent in vitro study was performed on human skin explants from a donor to evaluate the effect of the melon concentrate at different levels including on the sunburn cells formation and on the endogenous antioxidant enzymes and its influence on melanin. Clinical study results demonstrate that melon concentrate application and/or supplementation increased MED. It also increased the endogenous antioxidant enzymes and reduced sunburn cells and melanin level on irradiated skin explants. Therefore, it is suggested that melon concentrate administration (oral and/or topical) could be a useful strategy for photoprotection due to its antioxidant properties.
Article
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The present study investigated the anti-aging effects of pomegranate juice concentrated powder (PCP) in hairless mice following 15 weeks of UVB irradiation (three times a week; 0.18 J/cm²). Skin moisturizing effects were evaluated through skin water, collagen type I and hyaluronan contents, as well as collagen type I and hyaluronan synthesis-related transcript levels. Wrinkle formation and edema scores (skin weights) were also assessed, along with skin matrix metalloproteinase (MMP)-1, MMP-9 and MMP-13 transcript levels. To determine the anti-inflammatory effects of PCP, myeloperoxidase (MPO) activity, interleukin (IL)-1β and IL-10 contents were observed. Caspase-3 and cleaved poly(ADP-ribose) polymerase (PARP) were used as an apoptotic index in epidermal keratinocytes. To determine the anti-oxidative effects of PCP, nitrotyrosine and 4-hydroxynonenal immunoreactive cells were detected and glutathione (GSH) content, malondialdehyde levels, superoxide anion production, Nox2, and GSH reductase mRNA expression were all measured. The results indicated that skin wrinkles induced by photoaging were significantly reduced by PCP, whereas skin water contents, collagen type I and hyaluronan contents all increased. Furthermore, IL-1β levels in the PCP-treated groups were lower than those in the UVB-exposed control group. UVB-induced GSH depletion was also inhibited by PCP. Taken together, the results of the current study suggest that PCP has favorable protective effects against UVB-induced photoaging through anti-apoptotic effects, MMP activity inhibition and ECM (COL1 and hyaluronan) synthesis-related moisturizing, anti-inflammatory and anti-oxidative effects.
Article
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Acne is a common skin condition with sebum overproduction, hyperkeratosis, Propionibacterium acnes (P. acnes) and Staphylococcus aureus, and inflammation. Punica granatum (pomegranate) is well-known for its anti-inflammatory effects; however, few studies have discussed the anti-acne effects of pomegranate. In this study, we found that pomegranate extract (PG-E) significantly reduced P. acnes-induced edema in Wistar rat ears. Therefore, an evaluation platform using multiple pathogenic mechanisms of acne was established to explore the anti-acne effects of pomegranate. Results showed that PG-E inhibited bacterial growth and lipase activity. Through a bioguided-fractionation-isolation system, four hydrolysable tannins, punicalagin (1), punicalin (2), strictinin A (3), and granatin B (4), were isolated. Compounds 1 and 2 had greater anti-bacterial activities and anti-testosterone-induced HaCaT proliferative effects than the others. Compounds 1, 3, and 4 displayed lipase inhibitory effects. Compound 4 decreased cyclooxygenase-2 expression and downregulated prostaglandin E2 production in heat-killed P. acnes-treated RAW 246.7 cells. In conclusion, PG-E is abundant in hydrolysable tannins that display multiple anti-acne capacities, including anti-bacterial, anti-lipase, anti-keratinocyte proliferation, and anti-inflammatory actions. Hence, PG-E has great potential in the application of anti-acne and skin-care products, and punicalagin (1), the most effective component in PG-E, can be employed as a quality control marker.
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Background: The human body relies on several aging defense mechanisms (ADMs) to limit damage induced from pro-aging stressors (aging aggressors). However, such protective mechanisms can be compromised, leading to accelerated aging. The skin provides a model to probe the effects of an oral nutritional intervention on ADMs in response to ultraviolet radiation (UVR)-induced damage. Objective: To determine whether supplementation with a novel nutritional and phytonutrient blend could protect against UVR-induced skin damage and positively influence facial skin attributes and characteristics by bolstering ADMs. Methods: Thirty-six healthy, nonsmoking women (40-75 years) with Fitzpatrick skin types I and II were recruited. UVR-induced erythema and the number of apoptotic cells were determined before (pre-) and after 8-week (post-) supplementation. Other clinical variables included skin carotenoid concentrations, facial skin attributes and characteristics. Results: Eight-week supplementation led to protection against UVR-induced skin damage as evidenced by reductions in erythema at all three minimal erythema doses (MEDs) (9.1 to 7.4 [P = 0.10]; 15.8 to 13.6 [P = 0.02]; and 19.6 to 17.3 [P = 0.01] for one, two, and three MEDs and a reduction in the average number of apoptotic cells [11.3 to 5.3, P < 0.0001] pre- and post-supplementation, respectively). Skin carotenoid concentrations increased from 28 111 Raman intensity units to 38 472 (P < 0.0001) along with noticeable improvements in facial skin attributes and characteristics: elasticity, transepidermal water loss, radiance, texture, and overall appearance (all P < 0.05) following supplementation. Conclusion: Eight weeks of oral supplementation positively impacted ADMs resulting in protection against UVR-induced skin damage and improvements in facial skin attributes and characteristics.
Book
While one may not find ancient studies that substantiate the pomegranate’s curative and preventive qualities, the exalted status of this fruit goes back as far as the history of agriculture itself. Allusions to the pomegranate are readily found in the oldest cultures of the Indus Valley, ancient China, and classical Greece, as well as in the Old Testament. To modern scientists, the biochemistry of the pomegranate is as equally fascinating as its storied place in literature and religion. Providing an unprecedented compilation of scientific information, Pomegranates: Ancient Roots to Modern Medicine offers an exploration of the biochemistry, health effects, and cultivation of the pomegranate that is as authoritative as it is unparalleled. Featuring the contributions of a multidisciplinary and international team of prominent researchers, it presents the latest findings on the potential human health benefits of this exceptionally polyphenol-rich fruit. As the research indicates, the physiological effects of pomegranate juice constituents are remarkable in their preventive potential against two of the major chronic diseases of aging - heart disease and cancer. Many of the pioneering researchers responsible for initiating our newfound fascination with pomegranates discuss its biochemistry, detailing the location and action of the phytochemicals found in the fruit’s flesh, peels and seeds. They present evidence of the pomegranate’s impact on heart disease, including its ability to enhance nitric oxide production in endothelial cells. They also reveal the significant antiproliferative and proapoptotic effects attributed to the pomegranate in battling several different types of cancer cells, as well as its ability to retard tumor growth in animals. Recognizing that the pomegranate is only as valuable as it is available, the editors include a substantial section on commercialization and another on plant growth and improvement. These additions make this text as uniquely essential for botanists and agriculturists as it is for nutritionists, cancer researchers, natural product chemists, botanical supplement producers and consumers, and pharmacognosists seeking to evaluate both the pomegranate’s legacy and future as a powerful natural healing agent.
Article
Background: Ultraviolet radiation (UVR) is a well-known factor in skin aging and pigmentation, and daily exposure to subcytotoxic doses of UVR might accelerate senescence and senescence-associated phenomena in human melanocytes. Objective: To establish an in vitro melanocyte model to mimic the conditions of repeated exposure to subcytotoxic doses of UVB irradiation and to investigate key factor(s) for melanocyte senescence and senescence-associated phenomena. Methods: Human epidermal melanocytes were exposed twice with 20 mJ/cm2UVB over a 24-h interval and subsequently cultivated for 2 weeks. Senescent phenotypes were addressed morphologically, and by measuring the senescence-associated β-galactosidase (SA-β-Gal) activity, cell proliferation capacity with cell cycle analysis, and melanin content. Results: The established protocol successfully induced melanocyte senescence, and senescent melanocytes accompanied hyperpigmentation. Prolonged expression of p53 was responsible for melanocyte senescence and hyperpigmentation, and treatment with the p53-inhibitor pifithrin-α at 2-weeks post-UVB irradiation, but not at 48 h, significantly reduced melanin content along with decreases in tyrosinase levels. Conclusion: Melanocyte senescence model will be useful for studying the long-term effects of UVB irradiation and pigmentation relevant to physiological photoaging, and screening compounds effective for senescence-associated p53-mediated pigmentation.
Article
The analysis of pomegranate phenolic compounds belonging to different classes in different fruit parts was performed by high-performance liquid chromatography coupled with photodiode array and mass spectrometry detection. Two different separation methods were optimized for the analysis of anthocyanins and hydrolyzable tannins along with phenolic acids and flavonoids. Two C18 columns, core–shell and fully porous particle stationary phases, were used. The parameters for separation of phenolic compounds were optimized considering chromatographic resolution and analysis time. Thirty-five phenolic compounds were found, and 28 of them were tentatively identified as belonging to four different phenolic compound classes; namely, anthocyanins, phenolic acids, hydrolyzable tannins, and flavonoids. Quantitative analysis was performed with a mixture of nine phenolic compounds belonging to phenolic compound classes representative of pomegranate. The method was then fully validated in terms of retention time precision, expressed as the relative standard deviation, limit of detection, limit of quantification, and linearity range. Phenolic compounds were analyzed directly in pomegranate juice, and after solvent extraction with a mixture of water and methanol with a small percentage of acid in peel and pulp samples. The accuracy of the extraction method was also assessed, and satisfactory values were obtained. Finally, the method was used to study identified analytes in pomegranate juice, peel, and pulp of six different Italian varieties and one international variety. Differences in phenolic compound profiles among the different pomegranate parts were observed. Pomegranate peel samples showed a high concentration of phenolic compounds, ellagitannins being the most abundant ones, with respect to pulp and juice samples for each variety. With the same samples, total phenols and antioxidant activity were evaluated through colorimetric assays, and the results were correlated among them.
Article
Skin microorganisms have adapted to utilize the sparse nutrients available on the skin Many cutaneous microorganisms can produce molecules that inhibit the colonization of other microorganisms or alter their behaviour The skin microbiota of a healthy adult remains stable over time, despite environmental perturbations Shotgun metagenomics provides greater resolution than traditional amplicon sequencing, enabling surveys of the skin microbiota at the kingdom, species, strain or gene level Skin microorganisms have important roles in educating the innate and adaptive arms of the cutaneous immune system Some skin diseases are associated with an altered microbial state; reversion of this dysbiosis may help prevent and/or treat the disease
Article
Humans have become exposed to another form of a trait which is ultraviolet B (UVB) radiation reaching the earth's surface. This has become a major source of oxidative stress that ultimately leads to inflammation, DNA damage, photoaging and pigmentation disorders etc. Although several studies have shown the photo-protective role of different grape parts like the fruits and seeds, little or no data demonstrating the in vivo photo-protective role of grape stem, which is the most discarded part of the grape are available. We evaluated the protective influence of grape stem extract against UVB-induced oxidative damage in C57BL mice characterized by epidermal hyperplasia, pigmentation, collagen degradation and inflammation. Grape stem extract was administered topically 1 week before UVB irradiation (120 mJ/cm²) and continued until the termination of the experiment. A group of non-irradiated mice and a group of irradiated mice topically administered with propylene were used as a negative and positive control. Epidermal thickness, pigmentation, erythema, mast cell and neutrophil infiltration, collagen degradation and COX-2, Nrf2, and HO-1 expressions were evaluated. Grape stem extract markedly recovered skin damage induced by the UVB radiation through the prevention of epidermal hyperplasia, pigmentation, erythema, mast cell and neutrophil infiltrations, collagen degradation and COX-2, Nrf2, and HO-1 expressions. Our study demonstrated for the first time in C57BL mice that grape stem extract reduces UVB-induced oxidative damage and hence can play a protective role in skin photo-damage.
Article
Microbes inhabiting the phyllosphere encounter harmful ultraviolet rays, and must develop adaptive strategies against this irradiation. In this study, we screened bacterial isolates originating from the phyllosphere of various plants which harbored absorbers of ultraviolet A (UVA), a wavelength range which is recognized as harmful to human skin. Of the 200 phyllosphere bacterial isolates we screened, methanol extracts from bacterial cells of seventeen isolates absorbed wavelengths in the range of 315–400 nm. All of the UVA-absorbing strains belonged to Methylobacterium species based on 16S ribosomal RNA gene sequences, suggesting that cells of this bacterial genus contain specific UVA-absorbing compounds. When cells of a representative Methylobacterium strain were extracted using various solvents, UVA absorption was observed in the extracts obtained using several aqueous solvents, indicating that the UVA-absorbing compounds were highly polar. A compound was purified using solid columns and HPLC separation, and comparative analysis revealed that the absorption strength and spectrum of the compound were similar to those of the known UVA filter, avobenzone. The compound was also verified to be stable under UVA exposure for at least 480 min. Based on these results, the UVA-absorbing compound harbored by Methylobacterium has potential to be used as a novel sunscreen ingredient.