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Jungle Honey Enhances Immune Function and Antitumor Activity


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Jungle honey (JH) is collected from timber and blossom by wild honey bees that live in the tropical forest of Nigeria. JH is used as a traditional medicine for colds, skin inflammation and burn wounds as well as general health care. However, the effects of JH on immune functions are not clearly known. Therefore, we investigated the effects of JH on immune functions and antitumor activity in mice. Female C57BL/6 mice were injected with JH (1 mg/mouse/day, seven times intra-peritoneal). After seven injections, peritoneal cells (PC) were obtained. Antitumor activity was assessed by growth of Lewis Lung Carcinoma/2 (LL/2) cells. PC numbers were increased in JH-injected mice compared to control mice. In Dot Plot analysis by FACS, a new cell population appeared in JH-injected mice. The percent of Gr-1 surface antigen and the intensity of Gr-1 antigen expression of PC were increased in JH-injected mice. The new cell population was neutrophils. JH possessed chemotactic activity for neutrophils. Tumor incidence and weight were decreased in JH-injected mice. The ratio of reactive oxygen species (ROS) producing cells was increased in JH-injected mice. The effective component in JH was fractionized by gel filtration using HPLC and had an approximate molecular weight (MW) of 261. These results suggest that neutrophils induced by JH possess potent antitumor activity mediated by ROS and the effective immune component of JH is substrate of MW 261.
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Hindawi Publishing Corporation
Evidence-Based Complementary and Alternative Medicine
Volume 2011, Article ID 908743, 8pages
Original Article
Jungle Honey Enhances Immune Function and
Antitumor Activity
Miki Fukuda,1Kengo Kobayashi,1Yuri ko Hi r o n o,1Mayuko Miyagawa,1Takahiro Ishida,1
Emenike C. Ejiogu,2Masaharu Sawai,3Kent E. Pinkerton,4and Minoru Takeuchi1
1Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Kyoto, Japan
2Origins Japan Co. Ltd, Nagano, Japan
3Takara Shuzo Co. Ltd, Kyoto, Motoyama, Kamigamo, Kita-ku, Kyoto 603-8555, Japan
4Center for Health and the Environment, University of California Davis, California, USA
Correspondence should be addressed to Minoru Takeuchi,
Received 16 June 2008; Accepted 17 December 2008
Copyright © 2011 Miki Fukuda et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Jungle honey (JH) is collected from timber and blossom by wild honey bees that live in the tropical forest of Nigeria. JH is used as
a traditional medicine for colds, skin inflammation and burn wounds as well as general health care. However, the eects of JH on
immune functions are not clearly known. Therefore, we investigated the eects of JH on immune functions and antitumor activity
in mice. Female C57BL/6 mice were injected with JH (1mg/mouse/day, seven times intra-peritoneal). After seven injections,
peritoneal cells (PC) were obtained. Antitumor activity was assessed by growth of Lewis Lung Carcinoma/2 (LL/2) cells. PC
numbers were increased in JH-injected mice compared to control mice. In Dot Plot analysis by FACS, a new cell population
appeared in JH-injected mice. The percent of Gr-1 surface antigen and the intensity of Gr-1 antigen expression of PC were increased
in JH-injected mice. The new cell population was neutrophils. JH possessed chemotactic activity for neutrophils. Tumor incidence
and weight were decreased in JH-injected mice. The ratio of reactive oxygen species (ROS) producing cells was increased in JH-
injected mice. The eective component in JH was fractionized by gel filtration using HPLC and had an approximate molecular
weight (MW) of 261. These results suggest that neutrophils induced by JH possess potent antitumor activity mediated by ROS and
the eective immune component of JH is substrate of MW 261.
1. Introduction
Natural products are known to have biological activity,
and we have previously investigated the eect of natural
products on immune function [1,2]. Honey contains various
vitamins, minerals and amino acids as well as glucose and
fructose and is popular as a natural food [36]. There is a
wide variety of honey (Manuka honey, Pasture honey, Jelly
bush honey and Jungle honey, etc.), and the varieties are
due to components of the flower sources. Honey is used
not only as natural food but also as traditional medicine for
health care, in beauty products and antiinflammatory skin
care. One variety, Jungle honey, is collected from timber and
blossom by wild honeybees that live in the tropical forest
of Nigeria. Jungle honey is used as traditional medicine or
preventive medicine to treat colds, skin inflammation and
burn wounds as well as for general health care.
It is generally known that honey has antibacterial activity
that has been reported to be due to its high osmolarity,
acidity and presence of hydrogen peroxide and unidentified
substances from floral sources [711]. It has been reported
that flavonoid and phenol acid show antibacterial activity
[1214]. It was reported in a clinical experiment that when
wound infected with gram positive and gram negative
bacteria were treated with honey, infection was more quickly
eradicated [1517].
It has been reported that Manuka honey increased IL-
1β,IL-6,andTNF-αproduction from Mono Mac6 cells or
human monocytes [18,19], and the active component was
5.8 kDa, which increased production of these cytokines via
TLR4 [20]. In addition, it was reported that oral intake
of honey augmented antibody productions in primary and
secondary immune responses against thymus-dependent and
thymus-independent antigens [21].
2 Evidence-Based Complementary and Alternative Medicine
Honey may provide the basis for the development of
novel therapeutics for patients with wounds. Therefore,
the purpose of this study was to investigate the eects of
Jungle honey on immune function and antitumor activity in
2. Methods
2.1. Preparation of Jungle Honey. Jungle honey (JH) was a gift
from Nihon origins Co. Ltd (Nagano, Japan). JH was har-
vested in the forest areas around the Nsukka area of Enugu
state, Nigeria [22]. Generally, the bees were Apis mellifera
adonsonii. The main plant species that the bees collected
from nectar were Pentaclethra macrophylla,Chrysophyllum
albidum and Milicia excela. Briefly, the components of JH
in 100 g were 900 mg protein, 1400 mg glucon acid, 657 mg
amino acid, 213 mg mineral and 3.46 mg vitamin. JH was
dissolved with distilled water, freeze dried and then adjusted
to 10 mg/mL with PBS(). JH was steriled by 0.22-μm
filtration (Millipore, MA, USA) and then stored at 4Cbefore
use. The endotoxin unit of JH was found to be 2.7EU/mL
using the limulus amebocyte lysate assay kit (Canbrex, MD,
USA). The mice did not immunologically respond to this
unit of endotoxin.
2.2. Mice. Female C57BL/6 mice were used at 8–10 weeks.
Ten mice were used in each group. Mice were obtained
from Japan SLC (Shizuoka, Japan). They were housed in
transparent plastic cages with stainless wire lids in the animal
facility of Kyoto Sangyo University (Kyoto, Japan). They were
maintained under standard conditions, with a dark period
from 8 pm to 8 am, and water and food were provided ad libi-
tum. This study was approved by the committee for animals
in Kyoto Sangyo University. Mice were intraperitoneally (i.p.)
injected seven times with JH at a dose of 1 mg/mouse/day.
Control mice received PBS()[Ca
2+-free Dulbecco’s
phosphate buered saline (Nissui Pharmaceutical, Tokyo,
2.3. Analysis of Peritoneal Cells (PC). Peritoneal cells were
analyzed using Fluorescence Activated cell Sorter (FACS)
Calibur (Becton-Dickinson, CA, USA). After seven injections
of JH, PC were collected by peritoneal lavage from the
mice with cold PBS. PC were pooled into plastic tubes
and centrifuged at 185 g for 10 min. The pelletted cells
were resuspended at 1 ×106cells/mL in FACS buer (PBS
containing 100 μg/mL CaCl2/MgCl2, 0.01% sodium azide
and 1% FCS). The newly appearing cell population found via
Dot Plot analysis after JH exposure was sorted by FACS. The
sorted cell population was suspended at 5 ×105cells/mL in
R(+) (RPMI1640 containing 10% fetal calf serum, 100 U/mL
penicillin, 100 μg/mL streptomycin). The cell suspension
(200 μLof5×105cells/mL) was put on a slide glass using
Cyto Spin and centrifuged at 185 g for 5 min. After the slide
glasses were dried in a dryer, the cells were fixed for 3min by
methanol, followed by Giemsa stain. Giemsa-stained samples
were observed using a light microscope.
2.4. Chemotaxis Assay Using EZ-TAXIScan. Neutrophils were
obtained from guinea pig peripheral blood. Blood was
diluted twice with PBS. To precipitate red blood cells,
peripheral blood was added in equal parts to 3.5% Dextran
in saline and incubated at room temperature for 30 min.
The leucocyte-rich supernatant was centrifuged at 400 g
for 30 min on a Ficoll-Paque Plus (GE Healthcare, Tokyo,
Japan) density gradient. The pellet was hemolyzed by
hypotonic lysis. Fractionated neutrophils were centrifuged
at 185 g for 10 min and resuspended at 2 ×106cells/mL in
R(+)(RPMI1640 containing 0.1% bovine serum albumin,
HEPES). A chemotaxis assay for neutrophils was evaluated
with EZ-TAXIScan. Time-lapse images of neutrophils during
chemotaxis were obtained using EZ-TAXIScan equipped
with a six channel chamber (GE Healthcare). This chamber
consists of an etched silicon substrate and a flat glass plate,
both of which form two compartments with a 4-μmdeep
microchannel. Neutrophils (1 μLof2×106cells/mL) were
put into a hole with which the device is held together
with a stainless holder, and 1μLof10
methionyl-leucyl-phenylalanine (fMLP) or 1 mg/mL JH was
put into a contra-hole. The holder assembly was filled with
R(+) and incubated for 30 min at 37C. A charge-coupled
device (CCD) camera was used to record the migration of
neutrophils toward the high concentration of each sample.
2.5. Antitumor Activity. Lewis Lung Carcinoma/2 (LL/2)
cells were used as tumor cells. LL/2 cells were maintained in
a 10-cm dish (BD Falcon, CA, USA) at 2- to 3-day intervals
using MEM(+) [D-MEM (Nacalai tesque, Kyoto, Japan)
containing 10% fetal calf serum, 100 U/mL penicillin and
100 μg/mL streptomycin]. After 2-3 days, LL/2 were obtained
with trypsin-EDTA (0.25% trypsin : 0.02% EDTA =1:1,
Nacalai tesque) and washed with MEM()(D-MEM con-
taining 100 U/mL penicillin and 100 μg/mL streptomycin).
Hemocytometer and trypan blue dye exclusion testing were
used to determine LL/2 total number and viability. Mice
injected with JH or PBS() were inoculated intraperitoneally
with LL/2 (4 ×105cells/0.2 mL/mouse). After 4 weeks,
antitumor activity was evaluated by tumor incidence and
weight. Tumor tissues were fixed in 10% neutral buered
formalin fixative and paran embedded. Sections (4 μm)
were stained with H&E.
2.6. Production of Reactive Oxygen Species. PC (1 ×
105cells/100 μL) were incubated with Hydroethidine (HE,
Polysciences, PA, USA, final concentration 10 μM) or 2,7-
Dichlorofluorescin diacetate (DCFH-DA, Sigma, MO, USA,
final concentration 20 mM). After a shaking incubation at
37C for 30 min, the cells were washed twice and resus-
pended in 200 μL of PBS(+) (Dulbecco’s phosphate buered
saline) and then analyzed using FACS.
2.7. Fractionation of Jungle Honey. JH (100 mg/mL) was
fractionized from Fr. 1 to Fr. 5 by gel filtration using a
Shodex OHpak SB-802 HQ column and HPLC (LC-20AD,
RID-10A, SPD-20A, CB-20A, Simazu, Japan). Elution was
Evidence-Based Complementary and Alternative Medicine 3
carried out with PBS()ataflowrateof1mL/minfor
30 min. Standards curves were traced using polyethylene
glycols, which had MW of 3930, 1020, and 106 (Polymer
Laboratories, Germany), and LCsolution GPC (Shimau),
under the same conditions in HPLC. The MW of JH was
estimated using the polyethylene glycol standard curves.
Each fraction of JH was freeze dried and then adjusted to a
concentration of 10 mg/mL with PBS().
2.8. Expression of IL-1βmRNA. Aliquots of obtained PC
(1 ×105cells/100 μL/well) were cultured with or without
JH fractions (final concentration 500 μg/mL) in 96-well flat
bottom culture plates (Becton-Dickinson, MA, USA) at
2. After 24 h, total RNA was iso-
lated by acid guanidinium thiocyanate-phenol-chloroform
assay. Total RNA was transcribed to cDNA with MLV
reverse transcriptase (Invitrogen, CA, USA). Oligonucleotide
primers were used from published cDNA sequences of IL-
1β(250 bp) and β-actin (268 bp) (house-keeping gene). PCR
was performed for 30 cycles using the following primer
pairs: β-actin sense (5-GCATTGTTACCAACTGGGAC-3)
and β-actin antisense (5-TCTCCGGAGTCCATCACAAT-
3); IL-1βsense (5-AGCTACCTGTGTCTTTCCCG-3)and
IL-1βantisense (5-GTCGTTGCTTGGTTCTCCTT-3). The
amplification profile consisted of denaturation at 94Cfor
30 s, primer annealing at 56C for 30 s and extension at
72C for 30 s. PCR products were visualized using ethidium
bromide after 8% polyacrylamide gel electrophoresis. Data
on the expression in IL-1βmRNA were quantified by Scion
2.9. Statistical Analysis. Allvaluesareexpressedasmean±
SE. Comparisons between control and JH-injected mice were
made with the Student’s t-test. Any P-values <.05 were
considered statistically significant.
3. Results
3.1. Increases of the Number of PC by Jungle Honey. The
number of PC was significantly (P<.001) increased in JH-
injected mice (5.13 ±0.28 ×106cells/mouse) compared to
control mice (1.17 ±0.11 ×106cells/mouse).
3.2. Induction of New Cell Populations of PC by Jungle
Honey. New cell populations in JH-injected mice were found
at FSC 120–400, SSC 200–800 by Dot Plot analysis of
FACS (Figure 1(b)) compared to control mice (Figure 1(a)).
An isolated, new cell population was found, as is shown
in Figure 1(c) and the isolated cells were found to be
neutrophils (Figure 1(d)) by Giemsa stain. The new cell
population in JH-injected mice was observed by a light
microscope and was identified as neutrophils by their
morphology (Figure 1(d)).
3.3. Enhancement of Chemotaxis for Neutrophil by Jungle
Honey. Forty neutrophils migrated in 30 min in the JH-
treated group compared to 13 neutrophils in the non-
treated group (Figure 2(a)-i, ii). The velocity of the migrating
neutrophils was 0.17 ±0.01 μm/s in the JH-treated group
and 0.04 ±0.01 μm/s in the non-treated group (Figure 2(b)).
The radian of neutrophils was 0.39 ±0.04 rad in JH-
treated group and 0.09 ±0.03 rad in the non-treated
group (Figure 2(b)). JH resulted in significantly (P<.001)
increased numbers, velocity, and radiation of migrated cells.
Therefore, JH showed chemotactic activity for neutrophils.
3.4. Inhibition of LL/2 Tumor Growth by Jungle Honey. The
incidence of LL/2 tumors was 20% in JH-injected mice
and 100% in control mice (Figure 3(a)). The mean tumor
weight was 0.02 ±0.02 g in JH-injected mice and 2.57 ±
1.05 g in control mice (Figure 3(b)). These results reveal
that JH inhibited tumor incidence and growth (Figure 4). In
histological findings of control tumor tissue, necrotic areas
were recognized (Figure 4(a)-i, iii), but there were few infil-
trations of neutrophils (Figure 4(a)-ii, iv). In JH injected-
tumor tissue, massive necrotic areas (Figure 4(b)-v) and
infiltration by many neutrophils were observed (Figure 4(b)-
vi). Hemorrhagic necrotic areas and a disassociation between
tumor cells were found (Figure 4(b)-vii, viii).
3.5. Increases of ROS Production in PC by Jungle Honey. The
ratio of control was 1.0. The ratio of O
2was 1.16 and the
ratio of H2O2was 1.13 in the JH treated group. The ratio
of O
2or H2O2producing cells were significantly (P<.001)
increased in JH-injected mice compared with control mice
(Figure 5). Therefore, Reactive Oxygen Species (ROS) may
be associated with antitumor activity.
3.6. Enhancement of IL-1βmRNA Expression in PC by Jungle
Honey Fractions. IL-1βmRNA expressions by JH and each
fraction from Fr. 1 to Fr. 5 were 0.94 ±0.06, 0.67 ±0.12, 1.11
±0.13, 0.42 ±0.05, 0.58 ±0.04 and 0.64 ±0.05, respectively
(Figure 6(a)). IL-1βmRNA expressions were significantly (P
<.001) increased by JH or Fr. 2 (Figure 6(b)).
4. Discussion
Jungle honey (JH) is collected from timber and blossom by
wild honeybees that live in the tropical forest of Nigeria,
where JH is used as traditional or preventive medicine for
colds, skin inflammation and burn wounds as well as general
health care. Therefore, we expected that JH would have
potential biological, especially immune, activity. Until now,
the eect of JH on immunomodulatory activity has been
relatively unknown. Therefore, we investigated the eects of
JH on immune function and antitumor activity in mice.
We found that the number of peritoneal cells (PC) was
increased 4-fold in JH-injected mice compared with con-
trol mice. This result suggests that JH induces cell migration.
Although the eects of other types of honey on PC numbers
are not yet reported, mice treated with other natural products
(i.e., hydroalcoholic extract from Chenopodium ambrosioides
or aqueous extract from Orbignya phalerata Mart)havebeen
shown to have increased PC as well [23,24].
To characterize the new cell population found after JH
treatment, we investigated surface antigens by FACS. In Dot
Plot analysis, a new cell population appeared in the region
4 Evidence-Based Complementary and Alternative Medicine
0 200 400
600 800 1000
(a) Control
0 200 400
600 800 1000
(b) Jungle honey
0 200 400
600 800 1000
(c) Isolated cell population
(d) Isolated cell population
Figure 1: Induction of new cell populations of PC by Jungle honey. (a) Control. (b) Jungle honey. (c) Isolated cell population. (d) Isolated
cell population.
of FSC 120–400 and SSC 200–800 in JH-injected mice. The
percent of Gr-1 surface antigen and the intensity of Gr-1
antigen expression of PC were increased in JH-injected mice
(data not shown). Moreover, the new cell population was
found to be neutrophils based on morphology. Although
the eects of honey on Dot Plot and cell surface antigen
of PC were not reported, it was shown that the number of
neutrophils was increased in treated mice with propolis [25].
This result agrees with our present report.
Our results showed that JH may have chemotactic
activity for neutrophils. Therefore, we investigated the
chemotactic activity of JH for neutrophils. The velocity and
direction of migration were increased by JH compared with
the control treatment. These results suggest that JH possesses
chemotactic activity for neutrophils. Although there are no
reports concerning chemotactic activity of honey, polysac-
charide from Ganoderma lucidum have chemotactic activity
for neutrophils in the Boyden chamber assay [26].
Because it was demonstrated that the number of PC
and migration of neutrophils were increased by JH, we
investigated antitumor activity by immune cells. LL/2 tumor
tumor antigen and inhibit the immune system as well as
human cancer [27]. The incidence and the mean weight of
LL/2 tumors were decreased in JH-injected mice compared
to control mice. These results suggest that JH has a preventive
eect on tumor growth. Tumor weight was found decreased
by royal jelly, propolis or polyphenol in propolis [2830].
The tumor tissue was infiltrated by many neutrophils at
massive necrotic areas in JH-injected mice. Therefore, it was
suggested that the neutrophils were involved with inhibition
of tumor growth.
To investigate the mechanism of antitumor activity by
JH, we examined ROS, a well-known antitumor factor.
The ratio of ROS produced by cells was increased in JH-
injected mice. Although the eect of honey on cellular
Evidence-Based Complementary and Alternative Medicine 5
(i) Control
(ii) Jungle honey
After 30 m in After 30 m in
(a) Image of neutrophil chemotaxis
Direction (Radian)
Veloc i t y ( μm/sec)
(b) Dot plots of velocity and direction of neutrophils
Figure 2: Enhancement of chemotactic activity for neutrophils by Jungle honey. (a) Image of neutrophil chemotaxis. (b) Dot plots of velocity
and direction of neutrophils. Up arrow: Migrated cells, filled circle: control, filled diamond: fMLP (106M), filled triangle: JH (1 mg/mL).
Incidence of LL/2 tumor (%)
0Control Jungle honey
(a) Incidence of LL/2 tumor
Tumo r w eig h t ( g)
Jungle honey
(b) Tumor weight
Figure 3: Inhibition of the incidence of LL/2 tumor (a) Tumor weight (a) by Jungle honey.
(i) (x200)
(iii) (x200)
(ii) (x400)
(iv) (x400)
(a) Control
(v) (x100)
(vii) (x200)
(vi) (x400)
(viii) (x400) H.E. Stain
(b) Jungle honey
Figure 4: Histological findings of LL/2 tumor by Jungle honey. (a) Control. (b) Jungle honey.
6 Evidence-Based Complementary and Alternative Medicine
0.9Control Jungle honey
1.2∗∗∗ ∗∗∗
Index of ROS production
Figure 5: Increases of ROS production in PC by Jungle honey. Open
square: O
Refractive index (RI)
0 5 10 15
Retention time
20 25 30
75000 Fr1 Fr2
M.W.3910 M.W.106
Fr3 Fr4 Fr5
(a) Fraction by gel filtration
Control Fr. 1 Fr. 2 Fr. 3 Fr. 4 Fr. 5
IL-1βmRNA expression/β-actin
(b) IL-1βmRNA expression
Figure 6: Enhancement of IL-1βmRNA expression in PC by Jungle
honey fractions. (a) Fraction by gel filtration. (b) IL-1βmRNA
ROS production has hereto been unreported, one study
reported that H2O2production was increased in PC treated
with extracts of G. lucidum or P. c o r nuc o p i a e [31]. This
result agrees with our present study. It was reported that
ROS produced by activated neutrophils has tumor cytotoxic
properties as well as preventive action against infection [32
35]. Since infiltration of many neutrophils was observed
at necrotic areas in JH injected-tumor tissue, there is a
possibility that antitumor activity by JH is due to production
of ROS by infiltrated neutrophils into tumor tissue.
JH was fractionized from Fr. 1 to Fr. 5 by gel filtration
using HPLC to identify the eective component in JH, and
IL-1βmRNA expressions in PC were assayed using each
fraction. IL-1βaugments immune response, functions and
migration capability of neutrophils. JH and Fr. 2 augmented
the expression of IL-1βmRNA in PC. Therefore, the eective
componentwasFr.2inJH,andtheeective component in
Fr. 2 was estimated to have a MW of 261. In contrast, it was
reported that the eective component of honey or royal jelly
was 55 kDa (Apalbumin-1), and the eective component of
Manuka honey was 5.8 kDa; these two components also were
found to increase TNF-αproduction [19,3638]. Therefore,
the active component of honey or royal jelly and Manuka
honey diers from the active component of JH.
Our results suggested that JH-induced neutrophils to the
peritoneal cavity, and the neutrophils were activated by IL-
1β, which was produced by JH stimulation. Then ROS from
activated neutrophils was associated with antitumor activity.
In addition, the eectivecomponentinJHwasfoundtohave
a MW of 261.
Grant-in-Aid for Scientific Research (C) in Japan Society for
the Promotion of Science (Grant no. 20500606).
The authors appreciate Dr Suzette Smiley-Jewell for her
eorts to provide technical assistance and suggestions in the
preparation of this manuscript.
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... An electronic paramagnetic resonance has also been used to study the MH's transitory action against superoxide anion radicals [43,44]. In these two studies, obtained outcomes provided that the alleviating role of MH could be very much due to the presence of methyl syringate [45]. Former in-vivo model experiments have proved the activity of MH in halting the damages caused by oxidation in young and middle-aged rodents hepatocytes [46]. ...
... Anticancer activity of MH against tested cell lines are surmised in Figure 2. A dosedependent anti-cancer potential was observed, as well as a substantial anticancer potential against the MCF-7 and A549 cell lines, and IC 50 values were as 9.05 and 9.37 mg/mL, respectively. Aside from its antibacterial and antioxidant characteristics, honey has been found in recent research to have features that could slow the multiplication of malignant cells [45,53,54]. The anti-cancerous activity can occur via the adoption of one of the following routes: (1) depolarizing the mitochondrial membrane of cancer cells, hence causing apoptosis, (2) cyclooxygenase-2 inhibition by components, such as flavonoids, (3) releasing cytotoxic H 2 O 2 and (4) scavenging the reactive oxygen species (ROS). ...
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Honey has a history of medical use and is known as bio-alternative therapy. This research assessed the phytochemical and biological activity of the medical grade manuka honey (MH). Gas chromatography–mass spectrometry (GC–MS) was chosen to investigate bioactive compounds of the MH. The DPPH and ABTS free radical scavenging and beta-carotene antioxidant activities as well as the antibacterial and antibiofilm effects against S. aureus, B. subtilis, E. coli and P. aeruginosa were all determined. Furthermore, to gauge anticancer properties of MH, a MTT assay was opted towards three cell lines, including HCT-116 (colon), A549 (lung) and MCF-7 (breast) cancer cells. The GC–MS analysis of the tested MH revealed the identification of various chemical constituents belonging to the fatty acids, phenols, and esters. The MH was found to have higher reducing power activity (DPPH IC50: 7.36; ABTS IC50: 4.49 mg/mL) than the beta-carotene bleaching power (IC50: 37.51 mg/mL). Similarly, the MH was noted to be more active against the planktonic and biofilm of Gram-positive bacteria. Furthermore, a dose-dependent anticancer potential was observed, although a significant anticancer potential was pointed out against the MCF-7 and A549 cell conforming to the IC50 values of 9.05 and 9.37 mg/mL, respectively. This study’s results have highlighted the MH’s chemical composition with significant bioactivities.
... Regular intake of honey has been shown to improve antibody production due to immune responses (Fukuda et al., 2011). It has been shown to induce antibody production during immune responses (thymus-dependent and independent antigens) in mice when exposed to sheep red blood cells and E. coli (Al-Waili and Haq, 2004). ...
... The pharmacological effects of honey can be the basis for developing novel therapeutics for patients suffering from cancer. Some jungle honey fragments were reported to induce chemotactic neutrophils and ROS, causing cell death (Fukuda et al., 2011). Also, a variety of Malaysian jungle honey has shown significant anticancer activity against human breast, oral, cervical, and osteosarcoma cell lines (Fauzi et al., 2011;Ghashm et al., 2010). ...
The Chemistry inside Spices and Herbs: Research and Development brings comprehensive information about the chemistry of spices and herbs with a focus on recent research in this field. The book is an extensive 2-part collection of 20 chapters contributed by experts in phytochemistry with the aim to give the reader deep knowledge about phytochemical constituents in herbal plants and their benefits. The contents include reviews on the biochemistry and biotechnology of spices and herbs, herbal medicines, biologically active compounds and their role in therapeutics among other topics. Chapters which highlight natural drugs and their role in different diseases and special plants of clinical significance are also included. Part II continues from the previous part with chapters on the treatment of skin diseases and oral problems. This part focuses on clinically important herbs such as turmeric, fenugreek, ashwagandha (Indian winter cherry), basil, Terminalia chebula (black myrobalan). In terms of phytochemicals, this part presents chapters that cover resveratrol, piperine and circumin.
... Various studies have been reported to investigate the anticancer benefits of different types of honey from different origins. The anticancer activity of honey has been demonstrated against various cancer cell lines and tissues, such as breast, prostate, colorectal, endometrial, and renal [98][99][100][101][102][103][104][105][106]. In general, the chemo-preventive properties of honey are consistent with its bioactive compounds, mostly quercetin, luteolin, chrysin, and esters of caffeic [107]. ...
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Osteoporosis and breast cancer are serious diseases that have become a significant socioeconomic burden. There are biochemical associations between the two disorders in terms of the amended function of estrogen, receptor activator of nuclear factor kappa beta ligand, oxidative stress, inflammation, and lipid accumulation. Honey as a functional food with high antioxidant and anti-inflammatory properties can contribute to the prevention of various diseases. Its health benefits are mainly related to the content of polyphenols. This review aims to summarize the current knowledge from in vitro, animal, and human studies on the use of honey as a potential therapeutic agent for osteoporosis and breast cancer. Preclinical studies have revealed a beneficial impact of honey on both bone health (microstructure, strength, oxidative stress) and breast tissue health (breast cancer cell proliferation and apoptosis, tumor growth rate, and volume). The limited number of clinical trials, especially in osteoporosis, indicates the need for further research to evaluate the potential benefits of honey in the treatment. Clinical studies related to breast cancer have revealed that honey is effective in increasing blood cell counts, interleukin-3 levels, and quality of life. In summary, honey may serve as a prospective therapeutic supplement for bone and breast tissue health.
... The diminution of cellular infiltration in the renal cortex of the honey/MSG-administered group reflects the potent immunomodulatory potential of honey [27] and its prominent role in the promotion of cellular immunity [28] . It reduces the relocation of these cells from the peripheral circulation because it possesses an anti-inflammatory role by suppressing the production of TNF-ɑ [26] . ...
Full-text available
Background: Monosodium glutamate (MSG) is a commercial food improver and is widely marketed as a flavor enhancer. It is now utilized in many processed foods and by most fast-food chains. Honey is a potent antioxidant that acts in the body against many diseases. Objective: The aim of the current study is to investigate honey's ameliorative effect on kidney damage initiated by monosodium glutamate in adult male rats. Moreover, different polyphenolic compounds in the crude honey were evaluated. Material and methods: Forty adult male albino rats were equally divided into four groups (N=6): The Control group was administered 1 mL of saline daily orally, the MSG group (30 g/kg on diet), the Honey group (2.5 g/kg body weight/day, orally) and Honey/MSG group as the previous for one month. Results: Urea, creatinine, and uric acid were significantly increased in the MSG group and decreased in the honey/MSG group compared to the control group. MSG markedly destructed glomeruli of the kidney and increased the immunoexpressing of nuclear erythroid-related factor 2 (Nrf2) and tumor necrosis factor-alpha (TNF-). Conclusions: Administration of crude honey attenuates and improves the kidney pathological changes induced by MSG.
... Nigerosa, a honey-derived sugar, is said to have immunostimulatory properties. 18 The production of TNF-α, IL-1, IL-6, and apalbumin 1 can be increased by a number of honeys, including Manuka, Royal Jelly, Grass, and Nigerian Jungle Honey. By raising levels of ascorbic acid, glutathione reductase, minerals, and immunological cells including eosinophils, monocytes, and lymphocytes in people, honey has positive effects. ...
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Due to their capacity to reduce acute inflammation by increasing the immune response, honey and its constituents are gaining attention as an efficient natural therapeutic. Studies have suggested that it may be able to treat a variety of chronic illnesses and conditions, including as bacterial and fungal infections, cardiac issues, diabetes, hypertension, and abnormalities of the lungs and heart. More significantly, honey has proven to have virucidal effects on a number of enveloped viruses, including the varicella-zoster virus, herpes simplex, influenza virus, and HIV. Honey may help patients with COVID-19, a disorder brought on by the SARS-CoV-2 enveloped virus, by boosting the host's immune system, treating comorbid conditions, and engaging in antiviral activities. Additionally, patients with COVID-19 are presently participating in a clinical trial of honey. We've attempted to condense the possible advantages of honey and its components in this review in terms of antibacterial properties, some chronic illnesses, and the host's immune system. Therefore, we have made an effort to develop a relationship with honey in order to treat COVID-19. In light of the COVID-19 epidemic, this review will be useful in reevaluating our understanding of the potential therapeutic benefits of honey. However, more in vitro and in vivo research is required to determine the effects of honey on SARS-CoV-2 replication and/or the host's immune system.
... This response is highlighted by the substantial increase in the total number of peritoneal cells and the recruitment of neutrophils. Our findings are consistent with previous studies showing that i.p. administration of jungle honey could induce recruitment of neutrophils into the peritoneal cavity of C57BL/6 mice, which were then able to limit tumor growth within the same site (47). The observed recruitment of neutrophils is directed through the secretion of chemokines which are primarily involved in the recruitment of inflammatory cells into the sites of injury (48). ...
Full-text available
Manuka honey (MH) is known for its wound-healing, anti-microbial, anti-oxidant and anti-tumor properties. However, there is conflicting evidence regarding the role of MH in inflammatory responses, with some studies highlighting its pro-inflammatory capacity and others showing that it has a predominantly anti-inflammatory activity. The current study is aimed at characterizing the immunomodulatory capacity of MH using both in vitro and in vivo approaches, focusing on the underlying mechanisms. Treatment of RAW 264.7 macrophages with 1% MH (w/v) resulted in a significant increase in the gene expression (~26-fold) and secretion (~27-fold) of tumor necrosis factor-alpha (TNF-α). Similarly, an increase was observed in the gene expression of other inflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS), as well as the chemokines; (C-X-C motif) ligand 2 (CXCL2) and (C-C) motif ligand 2 (CCL2). Using an in vivo model, intraperitoneal (i.p.) administration of MH in C57BL/6 mice elicited a peritoneal response characterized by a significant expansion in the number of peritoneal exudate cells (PECs), which was mainly due to a 35-fold increase in the recruitment of neutrophils. Importantly, this response was evident in toll-like receptor 4 (TLR4)-defective C3H/HeJ mice, indicating that the observed stimulatory effect occurs independently of TLR4 and unlikely to be mediated by any lipopolysaccharide (LPS) contaminant. MH administration also led to changes in the phenotypic expression and functional maturation of peritoneal macrophages, as evidenced by a shift towards the CD11blo F4/80lo phenotype and an increase in the expression of major histocompatibility complex (MHC) class II proteins. In contrast, the MH-initiated peritoneal response was largely abrogated in mice deficient in myeloid differentiation primary response 88 (MyD88) protein, a critical adaptor of most TLR signaling pathways. Thus, the current findings help to characterize the immunostimulatory properties of MH and their dependence on TLR signaling, and highlight the potential utility of MH as an immunomodulatory agent in a variety of disorders.
... It has been reported that honey stimulates B-lymphocytes and T-lymphocytes in cell culture to multiply and activate neutrophils, which induce cytokine production, such as, IL-1, IL-6, and TNF-α and apalbumin 1 (AP-1) [131]. A variety of honeys have been associated with the increase of immune responses mediators [132,133]. Tonks et al. [134] discovered a 5.8 kDA component of Manuka honey that stimulates the production of TNF-α in macrophages via TLR4, such as blockade suppress honey-mediated immunomodulatory effects. ...
Full-text available
Since ancestor times, honey has been used to promote human health due to its medicinal, and nutritious properties, mainly due to bioactive compounds present, such as phenolic compounds. The emergence of COVID-19, caused by the SARS-CoV-2 virus, led to the pursuit of solutions for the treatment of symptoms and/or disease. Honey has proven to be effective against viral infections, principally due to its potential antioxidant and anti-inflammatory activities that attenuate oxidative damage induced by pathogens, and by improving the immune system. Therefore, the aim of this review is to overview the abilities of honey to attenuate different COVID-19 symptoms, highlighting the mechanisms associated with these actions and relating the with the different bioactive compounds present. A brief, detailed approach to SARS-CoV-2 mechanism of action is first overviewed to allow readers a deep understanding. Additionally, the compounds and beneficial properties of honey, and its previously application in other similar diseases, are detailed in depth. Despite the already reported efficacy of honey against different viruses and their complications, further studies are urgently needed to explain the molecular mechanisms of activity against COVID-19 and, most importantly, clinical trials enrolling COVID-19 patients.
... Honey provides various nutritional benefits such as antiinflammatory (22), anti-hypertensive, and cardioprotective properties (23,24), as well as antioxidative effects (25)(26)(27). It also possesses antibacterial (28), antifungal (29), antiviral (30), and antitumor (31) activities. Honey has also demonstrated the anti-obesity effects in various studies (1,(32)(33)(34)(35). ...
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Obesity is a metabolic disorder that has become critically prevalent throughout the world. Obesity has been linked to other chronic diseases such as diabetes mellitus, cardiovascular diseases and cancer. Natural products such as honey have been investigated for their potential effect on obesity. Hence, this study systematically reviewed the recent literature concerning the effects of honey on obesity in obese animal models and in people with obesity. The Ovid MEDLINE, PubMed, Scopus, Web of Science and Google Scholar electronic databases were searched for relevant articles. A total of 130 relevant articles were obtained from the initial search. Following a thorough screening, nine articles were selected for data extraction, including six animal studies and three clinical trials. In most of the animal studies, honey demonstrated an anti-obesity effect by reducing body weight, body fat composition and adipocyte size, among others. However, supplementation of honey in clinical trials showed conflicting results. Even though honey supplementation did not demonstrate any weight-reducing effect in some of the clinical trials, none of the trials showed that honey increases body weight. However, the results should be interpreted with caution as most of the studies involved animal models and there is a limited number of high quality, randomized, controlled clinical trials. Systematic Review Registration PROSPERO, identifier 10.37766/inplasy2022.6.0038.
... According to certain studies, honey (manuka) possesses anti-proliferative properties against cancer cells due to its high content of polyphenols and flavonoids [36][37][38][39]. ...
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Manuka honey originates from the New Zealand manuka tree (Leptospermum scoparium). It has been recognized for its anti-bacterial and wound-healing activity. Antibacterial activity of manuka honey is due to reactive methylglyoxal (MG) as main constituent. It is a complex mixture of carbohydrates, fatty acids, proteins, vitamins and minerals containing numerous kinds of phytochemicals with high phenolic and flavonoid content. It have also antiulcer property, cancer and skin property. Also used medicinally all over the world. Manuka honey so called unique manuka honey with the presence of methylglyoxal.
The present study was performed to clarify the possible causes of the antimicrobial activity of honey A sugar solution resembling honey in its high sugar content was made. The antimicrobial activities of both honey and this solution towards 21 types of bacteria and two types of fungi were examined. The results achieved by both were compared. The difference between them indicated the presence of antimicrobial substance(s) in honey The kinds of antimicrobial substances (inhibines) in honey are discussed. Hydrogen peroxide is not the only inhibine in honey. In fact, inhibines in honey include many other substances. Two important classes of these inhibines are the flavonoids and the phenolic acids. Flavonoids have often been extracted from honey previously. In this study two phenolic acids (caffeic acid and ferulic acid) were extracted from honey for the first time.
Babassu is the popular name of Orbignya phalerata Mart. [Arecaceae (Palmae)], which fruits mesocarp has been used in Brazil as medicine for the treatment of pains, constipation, obesity, leukemia, rheumatism, ulcerations, tumors and inflammations. In this study, we investigated the effect of babassu mesocarp flour aqueous extract (BM) on C3H/HePas mice peritoneal cellular migration and macrophage activation by measuring the nitric oxide (NO), hydrogen peroxide (H2O2) and tumor necrosis factor (TNF) release, spreading activity and major histocompatibility complex (MHC) class II expression. Our results demonstrate that BM injected once ip in mice at 10 and 20 mg/kg increased the cellular influx to the peritoneal cavity, the MHC class II expression and the spreading ability, and also induced the production of NO, TNF and H2O2. The increase in NO-production and MHC expression was also observed after the addition of BM to resident macrophage cultures (100 mu g/ml). Thus, BM-treatment was able to activate peritoueal macrophages' in vitro and in vivo inducing the production of inflammatory and cytotoxic metabolites, which could justify the popular use of babassu mesocarp in the treatment of tumor diseases, but not in inflammatory pathologies. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
Although honey has been used as a traditional remedy for burns and wounds, the potential for its inclusion in mainstream medical care is not well recognized. Many studies have demonstrated that honey has antibacterial activity in vitro, and a small number of clinical case studies have shown that application of honey to severely infected cutaneous wounds is capable of clearing infection from the wound and improving tissue healing. The physicochemical properties (eg, osmotic effects and pH) of honey also aid in its antibacterial actions. Research has also indicated that honey may possess antiinflammatory activity and stimulate immune responses within a wound. The overall effect is to reduce infection and to enhance wound healing in burns, ulcers, and other cutaneous wounds. It is also known that honeys derived from particular floral sources in Australia and New Zealand (Leptospermum spp) have enhanced antibacterial activity, and these honeys have been approved for marketing as therapeutic honeys (Medihoney and Active Manuka honey). This review outlines what is known about the medical properties of honey and indicates the potential for honey to be incorporated into the management of a large number of wound types. (J WOCN 2002;29:295-300.)
This paper attempts, in an exploratory manner, to identify the various ways in which unsustainable beekeeping and honey hunting practices result in the loss of important multi-purpose agroforestry tree species in bee endemic parts of Southeastern Nigeria. Both qualitative and quantitative approaches (Rapid Rural Appraisal (RRA), community fora, focus group discussions, key informant interviews, and semi-structured interview schedules) were used in an interactive manner to collect data for this study from five randomly selected communities in Nsukka, Igbo-Etiti, Uzo-Uwani, Igbo-Eze South and Udenu Local Government Areas of Enugu State. Results show that beekeeping/honey hunting in the area is traditionally gender-specific occupations involving only male members of the households, while female members play an active role in processing, preserving, and marketing the products. The majority (69.6%) of the beekeepers/ honey hunters were within the age range of 31–50, and most (75.2%) of them did not receive secondary school education. Findings reveal that outright felling of some trees in order to extract honey, cutting tree trunks open and/or cutting down tree branches and setting surrounding bush on fire are among the major factors negatively impacting on the agroforestry of the area. Species of trees particularly at risk include Irvingia gabonensis, young Chlorophera excelsa, Raphia spp., Elaeis guineensis, Brachystegia eurycome, Dialium guineense, Erythrophleum guineese and Strychnos spinosa. Recommendations are proffered to improve harvesting practices for hive and other non-timber forest products in order to avert the erosion of natural resource base of the fragile farming ecosystem of the area.
The acids from clover honey were isolated by ion-exchange adsorption and separated by silicic acid partition chromatography and ion-exchange chromatography. The different acids were identified by paper chromatography with six solvent systems, infrared spectra of the sodium salts, and derivatives. Acids definitely identified were butyric, acetic, formic, lactic, succinic, pyroglutamic, malic, citric, and gluconic. Oxalic acid was tentatively identified.Gluconic acid, not citric acid as previously proposed, was the principal acid of honey.
The oligosaccharide fraction of samples of manuka (Leptospermum), heather (Calluna), clover (Trifolium) and beech honeydew (Nothofagus) honeys from New Zealand was separated from the monosaccharides and then analysed by high performance anion-exchange chromatography with pulsed amperometric detection (hpaec-pad). Significant oligosaccharide components of manuka honey were isomaltose (or maltulose), kojibiose, turanose (or gentiobiose), nigerose and maltose which was the major component. The composition of clover honey was identical to that of manuka, while heather honey differed from these two only because isomaltose was the major component. Beech honeydew honey was characterised by the complexity of the oligosaccharide composition. The trisaccharides melezitose and panose were the most abundant components. No differences were observed between the oligosaccharide compositions of manuka honeys which did or did not exhibit non-peroxide residual antibacterial activity. Manuka honey was shown to be derived from nectar and not honeydew as has been suggested.
The nonperoxide antibacterial activity of honey and honey fractions was tested withStaphylococcus aureusandMicrococcus luteusbacterial species. Antibacterial activity correlated significantly with the honey acidity but did not correlate with honey pH. There were small differences between the antibacterial activities of different honey types: rhododendron, eucalyptus and orange honeys had a relatively low activity, whereas dandelion, honeydew and rape honeys had a relatively higher activity. These results suggest that a part of the antibacterial activity might be of plant origin. However, the antibacterial activity of sugar-adulterated honeys was the same as that of control honeydew honeys produced in the same apiary suggesting that the major part of the antibacterial activity of honeydew honey is of bee origin.Ten different honeys were fractionated into four fractions using column chromatography or vacuum distillation: acidic; basic; nonvolatile, nonpolar; and volatile. The antibacterial activity of the different fractions tested was: acids > bases = nonpolar, nonvolatiles > volatiles. This order was the same using eitherStaph. aureusorMicrococcus luteusas test strains.An exception was manuka honey from New Zealand where almost the entire activity was found in the acidic fraction.
The present study was performed to clarify the possible causes of the antimicrobial activity of honey. A sugar solution resembling honey in its high sugar content was made. The antimicrobial activities of both honey and this solution towards 21 types of bacteria and two types of fungi were examined. The results achieved by both were compared. The difference between them indicated the presence of antimicrobial substance(s) in honey. The kinds of antimicrobial substances (inhibines) in honey are discussed. Hydrogen peroxide is not the only inhibine in honey. In fact, inhibines in honey include many other substances. Two important classes of these inhibines are the flavonoids and the phenolic acids. Flavonoids have often been extracted from honey previously. In this study two phenolic acids (caffeic acid and ferulic acid) were extracted from honey for the first time.