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Study on effect of anti-diarrheal medicinal plants on enteropathogenic Escherichia coli induced interleukin-8 secretion by intestinal epithelial cells

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Diarrhea is a major health concern in developing countries with enteropathogenic Escherichia coli (EPEC) being a leading cause of infantile diarrhea. Much of the pathology of EPEC infection is due to the inflammatory responses of infected intestinal epithelium through secretion of pro-inflammatory cytoki - nes such as interleukin (IL)-8. With medicinal plants gaining popularity as prospective antidiarrheal agents, we aimed to evaluate the effect of anti-diarrheal medicinal plants on secretion of IL-8 by epithelial cells in response to EPEC infection. The effect of the decoctions of four anti-diarrheal medicinal plants viz. Aegle marmelos , Cyperus rotundus , Psidium guajava and Zingiber officinale was studied on secretion of IL-8 by a human colon adenocarcinoma cell line, HT-29 infected with E. coli E2348/69. Two protocols were used viz. pre-incubation and post-incubation. The data obtained demonstrated that out of the four plants used, only P. guajava decreased secretion of IL-8 in the post-incubation protocol although in the pre-incubation protocol an increase was observed. A similar increase was seen with C. rotundus in the preincubation protocol. No effect on IL-8 secretion was observed with A. marmelos and Z. officinale in both protocols and with C. rotundus in the post-incubation protocol. The post-incubation protocol, in terms of clinical relevance, indicates the effect of the plant decoctions when used as treatment. Hence P. guajava may be effective in controlling the acute inflammatory response of the intestinal epithelial cells in response to EPEC infection. </p
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[page 64] [Alternative Medicine Studies 2011; 1:e16]
Study of effect of
anti-diarrheal medicinal plants
on enteropathogenic
Escherichia coli
induced
interleukin-8 secretion by
intestinal epithelial cells
Brijesh S.,1Pundarikakshudu Tetali,2
Tannaz J. Birdi1
1The Foundation for Medical Research,
84A, RG Thadani Marg, Worli, Mumbai,
2Naoroji Godrej Centre for Plant
Research, Lawkin Ltd. Campus,
Shindewadi, Shirwal, Satara, India
Abstract
Diarrhea is a major health concern in devel-
oping countries with enteropathogenic
Escherichia coli (EPEC) being a leading cause
of infantile diarrhea. Much of the pathology of
EPEC infection is due to the inflammatory
responses of infected intestinal epithelium
through secretion of pro-inflammatory cytoki -
nes such as interleukin (IL)-8. With medicinal
plants gaining popularity as prospective
antidiarrheal agents, we aimed to evaluate the
effect of anti-diarrheal medicinal plants on
secretion of IL-8 by epithelial cells in response
to EPEC infection. The effect of the decoctions
of four anti-diarrheal medicinal plants viz. Aegle
marmelos, Cyperus rotundus, Psidium guajava
and Zingiber officinale was studied on secretion
of IL-8 by a human colon adenocarcinoma cell
line, HT-29 infected with E. coli E2348/69. Two
protocols were used viz. pre-incubation and
post-incubation. The data obtained demonstrat-
ed that out of the four plants used, only P. guaja-
va decreased secretion of IL-8 in the post-incu-
bation protocol although in the pre-incubation
protocol an increase was observed. A similar
increase was seen with C. rotundus in the pre-
incubation protocol. No effect on IL-8 secretion
was observed with A. marmelos and Z. officinale
in both protocols and with C. rotundus in the
post-incubation protocol. The post-incubation
protocol, in terms of clinical relevance, indicates
the effect of the plant decoctions when used as
treatment. Hence P. guajava may be effective in
controlling the acute inflammatory response of
the intestinal epithelial cells in response to
EPEC infection.
Introduction
Diarrheal diseases are a major health con-
cern in developing countries with an estimated
1.8 million deaths per annum.1Despite improve-
ments in public health and economic well being,
it remains an important clinical problem in
developed countries as well.2It is estimated that
infectious diarrhea will remain a cause of global
health concern in the next 2-3 decades.3
Diarrhea is an etiologically diverse condi-
tion caused by a variety of enteric pathogens.4
E. coli is recognized to be a common cause of
gastroenteritis and accounts for nearly 30% of
total diarrheal pathogens in some regions.5
There are seven different categories of diar-
rheagenic E. coli strains based on epidemiolo-
gy, clinical syndromes, and virulence proper-
ties.6The enteropathogenic E. coli (EPEC) is
an important category that is a leading cause
of infantile diarrhea in developing countries.7
EPEC colonize the intestinal epithelial sur-
face and cause histopathological mucosal
changes, the physiological end results of which
are disruption of the intestinal epithelial barri-
er, alterations in intestinal transport, and
inflammation.8Much of the pathology of EPEC
infection has been linked to inflammatory
responses by the infected epithelium which
occurs through activation of the nuclear tran-
scription factor NF-κB. Since EPEC is a non-
invasive pathogen, NF-κB is activated by solu-
ble factors secreted or shed by the pathogen or
translocated type three secretory system-
dependent effectors.9Activation of NF-κB in
turn promotes the expression of proinflamma-
tory cytokines such as interleukin (IL)-810,11
which result in substantial recruitment of neu-
trophils and other PMNs to the site of in vivo
infection.12 Although the inflammatory
response is not the mechanism that initiates
EPEC mediated diarrhea, it may contribute to
the duration and severity of the diarrheal
response.8The PMNs and other inflammatory
cells recruited to the infected intestine also
cause considerable tissue damage through the
release of toxic inflammatory mediators.13
In recent years, medicinal plants have
gained popularity as prospective antidiarrheal
agents, with a large numbers of studies being
published in the past decade.14 Whilst a few
studies have reported antimicrobial activity, a
majority have focused on physiological diar-
rhea and thereby reporting their effect on
intestinal motility in experimental models.
Hence, we studied the activity of selected
antidiarrheal medicinal plants against patho-
genicity of infectious diarrhea, including
parameters such as colonization of epithelial
cells and production and action of enterotox-
ins.15-18 In the present study we aim to screen
medicinal plants for their effect on inflamma-
tory responses to infections by enteric
pathogens. These plants, if active, may prove to
be an effective therapy towards control of the
intestinal inflammation associated with infec-
tious forms of diarrhea.
Different approaches have been used to
analyze the generally anti-inflammatory poten-
tial of plants and plant-derived compounds
which include acute and chronic inflammatory
models. The acute models, which include both
in vitro and in vivo studies, are designed to
assess the effect of the plants on secretion of
pro-inflammatory cytokines such as IL-8,
changes in vascular permeability, leukocyte
migration and chemotaxis, macrophage activi-
ty, and measurement of rat paw edema.19
Therefore, in the present study we have evalu-
ated the in vitro activity of hot aqueous
extracts (decoction) of four selected antidiar-
rheal medicinal plants i.e. Aegle marmelos (L.)
Correa (Family Rutaceae), Cyperus rotundus
Linn., (Family Cyperaceae), Psidium guajava
L. (Family Myrtaceae) and Zingiber officinale
(Roscoe) (Family Zingiberaceae) against
secretion of the pro-inflammatory cytokine IL-
8 by HT-29 cells in response to EPEC infection.
These plants were chosen on the basis of an
ethonobotanical survey in the Parinche valley,
near Pune, Maharashtra, India.20 All four
plants are widely used in indigenous systems
of medicine across the world for their various
medicinal properties. They are widely used for
treatment of diarrhea and have been reported
to have immuno-modulatory properties includ-
ing anti-inflammatory activity.21-24
Alternative Medicine Studies 2011; volume 1:e16
Correspondence: Dr. Tannaz J. Birdi, Deputy
Director, The Foundation for Medical Research,
84A, RG Thadani Marg, Worli, Mumbai - 400018,
India.
Tel. +91.22.249.349.89 - Fax: +91.22.24932876
E-mail: fmr@fmrindia.org
Key words: Aegle marmelos, Cyperus rotundus,
Psidium guajava, Zingiber officinale, IL-8,
enteropathogenic Escherichia coli.
Contributions: BS carried out the laboratory stud-
ies, analyzed the data and prepared the manu-
script; PT collected the plant materials, authenti-
cated them and obtained voucher specimen num-
bers; TJB has overall responsibility for the study.
Conflict of interest: the authors report no con-
flicts of interest.
Received for publication: 17 May 2011.
Revision received: 28 September 2011.
Accepted for publication: 21 October 2011.
This work is licensed under a Creative Commons
Attribution NonCommercial 3.0 License (CC BY-
NC 3.0).
©Copyright Brijesh S. et al., 2011
Licensee PAGEPress, Italy
Alternative Medicine Studies 2011; 1:e16
doi:10.4081/ams.2011.e16
Non-commercial use only
[Alternative Medicine Studies 2011; 1:e16] [page 65]
Materials and Methods
Media, reagents, plastic ware and
instrumentation
The bacteriological media were purchased
from HiMedia laboratory, Mumbai, India.
Dulbecco’s modified Eagle medium (DMEM)
and fetal calf serum (FCS) were procured from
GibcoBRL, UK. The Quantikine human
CXCL8/IL-8 immunoassay kit was purchased
from R&D Systems, USA. All chemicals were
from SD Fine Chemicals, Mumbai. The 24-well
flat bottomed tissue culture plates were pur-
chased from Nunclon, Denmark, and the 55
mm diameter tissue culture plates were
obtained from Tarsons, Kolkata, India.
Bacterial strain
For the present study, EPEC strain E2348/69,
serotype 0127:H6 (kindly provided by Dr. S.
Knutton, Institute of Child Health, University
of Birmingham, Birmingham, UK) was used.
The bacterial strain was stored at -80°C in
brain heart infusion (BHI) containing 20%
glycerol. For each assay a frozen stock of the
bacterial strain was revived in BHI.
Cell culture
The human colon adenocarcinoma cell line,
HT-29, was obtained from the National Centre
for Cell Sciences, Pune, India. The cell line was
maintained by passage every 4-5 days in
DMEM supplemented with 10% FCS, at 37°C in
5% CO2atmosphere.
Plant material
The unripe fruits of A. marmelos, rhizomes of
C. rotundus and Z. officinale and the leaves of P.
guajava were collected from the Parinche valley,
about 53 km south east of the city of Pune in the
state of Maharashtra, India. The plant materials
were authenticated by Dr. P. Tetali, Naoroji
Godrej Centre for Plant Research (NGCPR).
Voucher specimens of A. marmelos, C. rotundus
and P. guajava were deposited at the Botanical
Survey of India (BSI), Western Circle, Pune,
Maharashtra, India, and that of Z. officinale at
the herbarium at the NGCPR (Table 1). The
plant materials were shade dried and powdered,
and stored at 4°C until used.
Preparation of extract
Crude aqueous extracts (decoctions) were
used for the study since this represents the
nearest form to traditional preparations. The
decoctions were prepared as described in the
Ayurvedic text:25 1 g of the powdered plant
materials were boiled in 16 mL double distilled
water till the volume reduced to 4 mL. The
decoctions were centrifuged and filtered
through a 0.22 μm membrane before use. To
replicate field conditions, each assay was per-
formed with freshly prepared decoctions. The
decoctions were diluted 1:1000, 1:100, 1:20 and
1:10 in DMEM for each experiment and have
been referred to as 0.1%, 1%, 5%, and 10%,
respectively throughout the text.
The dry weight of the decoctions was record-
ed to determine the concentration of the
extracts in each dilution. The qualitative phy-
tochemical analysis was performed using stan-
dard methods.26
IL-8 secretion by intestinal
epithelial cells in response to EPEC
infection
The effect of the decoctions on the inflamma-
tory response of intestinal epithelial cells to
infection by EPEC was measured by estimation
of IL-8 secreted by HT-29 cells in response to E.
coli E2348/69 supernatant by IL-8 enzyme
linked immunosorbent assay (ELISA).9Briefly,
E. coli E2348/69 was grown overnight in BHI at
37°C. The culture supernatant obtained by cen-
trifugation at 2,000 rpm was stored at -20°C till
used for the assay. For each assay, 0.2 mL of the
thawed culture supernatant of E. coli E2348/69
was added directly to HT-29 cells (5¥105/well)
grown overnight in 24-well tissue culture plates
in a total assay volume of 0.5 mL containing 0.3
mL DMEM with 10% FCS. Following incubation
for 18-20 h, the culture supernatant was collect-
ed and centrifuged at 14,000 rpm. The super-
natant were either immediately estimated for
IL-8 using a Quantikine IL-8 immunoassay kit
as per the manufacturer’s instructions or stored
at -20ºC for estimation of IL-8 at a later date.
The optical density was measured at 450 nm
(reference 540 nm) on an ELISA plate reader
(Labsystems, Finland).
The IL-8 assay was performed using two dif-
ferent protocols: pre-incubation protocol and
post-incubation protocol. In the pre-incubation
protocol the HT-29 cells were incubated with
different concentrations of the plant decoc-
tions for 3 h prior to addition of the EPEC
supernatant whereas in the post-incubation
protocol the plant decoctions were added into
the assay system 3 h after the addition of the
EPEC supernatant to the HT-29 cells.
Statistical analysis
The results have been expressed as the
mean ± standard error of the percentage val-
ues of the test groups relative to control
(100%) from three independent experiments.
Data were analyzed by analysis of variance
(ANOVA) and Dunnett’s post test. A value of
P0.05 was considered to be statistically sig-
nificant. The EC50 values, wherever applicable,
were calculated by non-linear regression
analysis using the equation for a sigmoid con-
centration-response curve. All statistical analy-
ses were performed using the software Prism
4.0 (GraphPad Software, Inc., USA).
Results
Phytochemical analysis
The percentage yield (mg/mL) for each
decoction with respect to the respective start-
ing material is shown in Table 1. The qualita-
tive phytochemical analysis revealed the pres-
ence of constituents such as carbohydrates,
flavonoids and tannins in the decoctions of all
four plants. While saponins were detected in
the decoctions of A. marmelos, C. rotundus and
P. guajava, phytosterols and anthraquinone
glycosides were detected in the decoction of A.
marmelos only (Table 2).
Effect on IL-8 secretion in response
to EPEC infection
The effect of the decoctions on EPEC
induced secretion of IL-8 by HT-29 cells is
shown in Figures 1-4. The baseline secretion
of IL-8 by HT-29 cells in the culture medium
was estimated to be 56.55±5.12 pg/mL where-
as the secretion of IL-8 in presence of EPEC
supernatant increased to 477.53±8.84 pg/mL
(represented as 100%). The decoctions of A.
marmelos and Z. officinale had no effect on
EPEC induced secretion of IL-8 by the HT-29
cells in either of the two protocols used
(Figures 1 and 4, respectively) whereas the
decoction of C. rotundus had no effect in the
post-incubation protocol (Figure 2). In com-
parison the decoction of P. guajava resulted in
a significant decrease (EC50 value
Article
Table 1. Details of the plant material.
Herbarium Botanical name Common name Family name Part Used % yield of
N. decoction
(w/w)
BSI-124675 A. marmelos Wood Apple, Rutaceae Unripe fruit pulp 20.4±2.11
Bengal Quince
BSI-124666 C. rotundus Nutgrass Cyperaceae Rhizomes 20.9±1.12
BSI-124672 P. guajava Guava Myrtaceae Leaves 10.8±0.5
NGCPR-642 Z. officinale Ginger Zingiberaceae Rhizomes 8.84±0.01
Non-commercial use only
[page 66] [Alternative Medicine Studies 2011; 1:e16]
2.09±0.78%) in EPEC induced IL-8 secretion in
the post-incubation protocol with a maximum
decrease at 10% concentration (Figure 3). On
the contrary, in the pre-incubation protocol the
decoctions of both C. rotundus and P. guajava
resulted in a significant increase in EPEC
induced IL-8 secretion (Figures 2 and 3,
respectively).
Discussion
The anti-inflammatory activity of four anti-
diarrheal medicinal plants used in traditional
medicine, i.e. A. marmelos, C. rotundus, P. gua-
java and Z. officinale were assessed against
EPEC induced secretion of IL-8 by intestinal
epithelial cells. Analysis of the results of the
study shows that the decoctions of A. marmelos
and Z. officinale had no effect on EPEC
induced IL-8 secretion by HT-29 cells in both
pre- and post-incubation protocols. On the
other hand, the IL-8 secretion by HT-29 cells
was increased in the pre-incubation protocol
by decoctions of C. rotundus and P. guajava and
inhibited in the post-incubation protocol by the
decoction of P. guajava.
Extracts from several plants have been
reported to inhibit IL-8 secretion from epithe-
lial cells.27-29 Our observations in the post-
incubation protocol on the inhibitory activity of
aqueous extract of P. guajava leaves on IL-8
secretion from epithelial cells confirm the
results reported by Peng et al.30 who used a
similar protocol. However, in contrast to the
observation made in the present study in
which the decoction of Z. officinale showed no
inhibition of IL-8 secretion, several studies
have reported on the inhibitory effect of the
ethanolic,31 50% ethanolic32 and methanolic33
extracts of Z. officinale on either IL-8 or IL-8
and NF-κB modulation in vivo and in vitro.
These differences in observations could be due
to the use of aqueous decoction in the present
study compared to the use of organic extracts
by other workers. Major bioactive constituents
from Z. officinale such as [6]-shogaol, [6]-
gingerol, [8]-gingerol, and [10]-gingerol,34
and constituents such as ar-curcumene and α-
pinene isolated from volatile oil,35 have also
been reported to have an inhibitory effect on
IL-8. However, these belong to the class of con-
stituents that are not soluble in water. In addi-
tion, the differences in the ecotype and/or vari-
ety of the Z. officinale used could also be
responsible for the observed difference in the
activity.
Inhibition of IL-8 secretion by epithelial
cells has been attributed to polyphenolic com-
pounds present in the extracts.36 Different
modes of action have been proposed for their
inhibitory activity on IL-8 secretion which
include: i) inhibition of inhibitory factor IKK
Article
Table 2. Results for the qualitative phytochemical analysis of the decoctions of the plants
used for the study.
Phytochemical Phytochemical Inference
Constituent Test A. marmelos C. rotundus P. guajava Z. officinale
Carbohydrates Molisch’s test ++++
Reducing sugars Fehling’s test ++++
Benedict’s test ++++
Starch Iodine test ++-+
Phytosterols Salkowski’s test +---
Cardiac glycosides Legal’s test ----
Anthraquinone Modified
glycosides Borntrager’s test +---
Saponins Foam test +++-
Flavonoids Shinoda’s test ++++
Tannins Ferric chloride test ++++
Lead acetate test ++++
Alkaloids Wagner’s test +-+-
Figure 1. Effect of A. marmelos on IL-8 secretion by HT-29 cells in response to EPEC
infection in the pre-incubation and the post-incubation protocols. Values represent mean
± standard error (n=3) of percentage secretion of IL-8 by HT-29 cells in presence of dif-
ferent concentrations of the decoction relative to control (100%).
Figure 2. Effect of C. rotundus on IL-8 secretion by HT-29 cells in response to EPEC
infection in the pre-incubation and the post-incubation protocols. Values represent mean
± standard error (n=3) of percentage secretion of IL-8 by HT-29 cells in presence of dif-
ferent concentrations of the decoction relative to control (100%). * P<0.05.
Non-commercial use only
[Alternative Medicine Studies 2011; 1:e16] [page 67]
activity, e.g. curcumin;37 and ii) inhibition of
MAPK activation, e.g. Punica granatum fruit
extract.38 It is known that different polypheno-
lic compounds influence different molecular
targets and pathways to exhibit their anti-
inflammatory activities.39 Though polyphenolic
compounds such as tannins and flavonoids
were detected in the crude decoctions of all
four plants, only the decoction of P. guajava
leaves inhibited IL-8 secretion by HT-29 cells.
Since EPEC is a non-invasive pathogen, the
activation of NF-κB has been linked to soluble
factors secreted or shed by the pathogen or
translocated type three secretory system-
dependent effectors.9Much of the pathology of
EPEC infection which is responsible for trig-
gering IL-8 release from epithelial cells has
been linked to flagellin, the flagellar structural
protein. Flagellin interaction with Toll-like
receptors (TLR)-5 results in activation of the
NF-κB which in turn promotes the expression
of IL-8.9The increase in IL-8 secretion by HT-
29 cells in the presence of the decoctions of C.
rotundus and P. guajava in the pre-incubation
protocol may be due to components that are
similar to flagellins structurally and/or func-
tionally which may be interacting with these
receptors and inducing expression of IL-8
mRNA. The effect could be similar to that
observed with green tea extract (GTE) which
was reported to induce de novo synthesis of IL-
8 in Caco-2 cells.40 Interestingly, Netsch et al.
reported that GTE, while inducing expression
of IL-8 mRNA, specifically inhibited its extra-
celluar secretion.40 Similarly, the decoction of
P. guajava, in addition to components that
induce expression of IL-8 mRNA, may also con-
tain constituents that may be specifically
inhibiting the extracellular secretion of IL-8.
This could probably be the reason for the inhi-
bition of IL-8 secretion observed in the post-
incubation protocol even when IL-8 expression
is induced. However, no decrease in extracellu-
lar secretion of IL-8 in the pre-incubation pro-
tocol following induction of IL-8 mRNA expres-
sion could be due to the removal of the decoc-
tion from the assay system prior to addition of
EPEC supernatant resulting in washing off the
components responsible for inhibiting the
extracellular secretion of IL-8.
It is interesting to note that the P. guajava
decoction had a biphasic effect on expression
of IL-8 mRNA in the pre-incubation protocol.
There was a dose dependent increase in secre-
tion of IL-8 with maximum secretion at a 5%
concentration. However, at a 10% concentra-
tion of decoction the IL-8 secretion was lesser
than that at 5%, though it was still much high-
er than the control. The observed biphasic
activity may probably be due to possible inter-
action between constituents of the crude
extract. Such biphasic activities of plant con-
stituents have been reported previously.41-44
However, as explained before, since the secre-
tion of IL-8 was specifically inhibited by cer-
tain plant constituents in the post-incubation
protocol in a dose dependent manner, the
biphasic effect on expression of IL-8 mRNA
was not apparent.
It is hypothesized that in terms of clinical
relevance the results of the pre-incubation pro-
tocol indicate a preventive effect of the plant
decoctions on the host whereas the post-incu-
bation protocol indicates the effect of the plant
decoctions when used as treatment. The
results, therefore, indicate that amongst the
plants tested, while none of them may be effec-
tive as preventive measures, P. guajava can be
used for the treatment of inflammatory diar-
rheal episodes. Our previous studies with
these four plants, however, have demonstrated
their efficacy against other parameters of
infectious forms of diarrhea, such as coloniza-
tion of epithelial cells, and production and
action of enterotoxins15-18 which still render
them effective for treatment of diarrhea.
In conclusion, it may be stated that of the
four antidiarrheal medicinal plants used in the
present study, only P. guajava is effective in
controlling the acute inflammatory response of
the intestinal epithelial cells in response to
EPEC infection.
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Figure 3. Effect of P. guajava on IL-8 secretion by HT-29 cells in response to EPEC infec-
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standard error (n=3) of percentage secretion of IL-8 by HT-29 cells in presence of differ-
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infection in the pre-incubation and the post-incubation protocols. Values represent mean
± standard error (n=3) of percentage secretion of IL-8 by HT-29 cells in presence of dif-
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Article
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... 33,34 Our previous studies also demonstrated the anti-diarrhoeal activity of guava leaves. 18,19,[35][36][37] These included the in vitro effect of GLD on the viability of S. flexneri and a reduction in the invading ability of the bacterium to HEp-2 cells. 18 Based on the promising results with the reference strain used, the current study was undertaken to elucidate the effect of guava leaves against drug-resistant clinical Shigella isolates. ...
Article
Background: Although shigellosis is self-limiting, antibiotics are recommended to minimize the severity of symptoms and reduce mortality rates. However, due to the increasing reports of antibiotic resistance, alternative approaches are needed to combat shigellosis. Interest for research on medicinal plants has increased in recent years, and hence, they can be explored to treat this infectious diarrhoea. Objective: To study the effect of Psidium guajava L. (guava) leaf decoction (GLD) on the antibiotic-resistant clinical isolates of Shigella spp. Materials and Methods: A total of 43 isolated Shigella spp. from diarrhoeal patients were used in this study. The effect of GLD on the bacterial viability was initially assessed. The isolates were divided into two categories: sensitive and resistant to GLD. For sensitive isolates, antibacterial activity of GLD was evaluated while for resistant strains, the ability of GLD for reducing the bacterial invasion of the HEp-2 cell line underwent an investigation. Results: Among the 43 Shigella isolates, GLD affected the growth of 23 strains. The invasion of 9 strains from the 20 remaining resistant isolates was unaffected. Although the number of isolates was less, the data suggested that isolates belonging to S. flexneri serogroup were more sensitive to GLD in comparison with other spp (i.e., sonnei, boydii, and dysenteriae). Conclusion: The results of this study revealed the efficacy of GLD against drug-resistant Shigella spp. and thus could be considered for the treatment of diarrhoea. GLD can be a cost-effective alternative to antibiotics.
... Guava leaves have been used globally in the treatment of gastrointestinal disorders [41,42]. Our work which included a clinical trial has confirmed that guava is a promising anti-diarrhoeal plant exhibiting a wide spectrum of activity [8,[43][44][45][46]. However, its functional components largely remained unknown, mainly due to chemical complexity and possible synergism. ...
Article
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Background Herbal medicines are fast gaining popularity. However, their acceptability by modern practitioners is low which is often due to lack of standardization. Several approaches towards standardization of herbals have been employed. The current study attempted to recognize key peaks from ¹ H NMR spectra which together would comprise of a spectral fingerprint relating to efficacy of Psidium guajava (guava) leaf extract as an antidiarrhoeal when a number of unidentified active principles are involved. Methods Ninety samples of guava leaves were collected from three locations over three seasons. Hydroalcoholic (water and ethanol, 50:50) extracts of these samples were prepared and their ¹ H NMR spectra were acquired. Spectra were also obtained for quercetin, ferulic acid and gallic acid as standards. Eight bioassays reflecting different stages of diarrhoeal pathogenesis were undertaken and based on pre-decided cut-offs, the extracts were classified as ‘good’ or ‘poor’ extracts. The bioactivity data was then correlated with the ¹ H NMR profiles using Regression or Orthogonal Partial Least Square-Discriminant Analysis (OPLS-DA). Results OPLS-DA showed seasonal and regional segregation of extracts. Significant models were established for seven bioassays, namely those for anti-bacterial activity against Shigella flexneri and Vibrio cholerae , adherence of E. coli , invasion of E. coli and S. flexneri and production and binding of toxin produced by V. cholerae . It was observed that none of the extracts were good or bad across all the bioassays. The spectral analysis showed multiple peaks correlating with a particular activity. Based on NMR and LC-MS/MS, it was noted that the extracts contained quercetin, ferulic acid and gallic acid. However, they did not correlate with the peaks that segregated extracts with good and poor activity. Conclusions The current study identified key peaks in ¹ H NMR spectra contributing to the anti-diarrhoeal activity of guava leaf extracts. The approach of using spectral fingerprinting employed in the present study can thus be used as a prototype towards standardization of plant extracts with respect to efficacy.
... Nigeria was estimated to have a total number of annual child deaths due to diarrhoea to be 151,700 [9]. Diarrhoea may be caused by a wide array of agents such as entero-pathogenic microorganisms (Shigella flexneri and Shigella dysenteriae, Staphylococcus aureus, Escherichia coli, Salmonella typhi and Candida albicans), alcohol, irritable bowel syndrome, bile salts, hormones, secretory tumors and intoxication [10,11]. Dependency on plants as medicine in controlling diseases is common among rural populace in Nigeria because of its relative safety and affordability compared with the cost of conventional medicines. ...
... Campylobacter species, S. aureus, and C. difficile), and parasitic (Cryptosporidium and Giardia) agents are important pathogens, although rotavirus is the major cause of infectious diarrhea, particularly among young children [4][5][6][7]. ...
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Diarrhea is a major health problem throughout the world and it has become more problematic in developing countries like Ethiopia. People, in several parts of the world, use different traditional medicines for treating diarrhea and it has been reported that the roots, leaves, and flowers of various species are used for the same purpose. In Ethiopia, for instance, Discopodium Penninervum is used for the treatment of diarrhea and also to control infection. The aim of the present study was, therefore, to evaluate the in vivo antidiarrheal and in vitro antimicrobial effect of Discopodium Penninervum in mice. For the antimicrobial activity test, four standard bacteria and disc diffusion method were used, while for antidiarrheal experiment, animals had been used, which were divided into 5 groups. The first group served as negative control and was administered with vehicle (0.2-0.3ml of distilled water). Groups two (D100), three (D200), and four (D400) were administered 100, 200, and 400 mg/kg of the extract, respectively. Group five served as positive control group and was administered with either loperamide (3mg/kg) for castor oil induced diarrhea and castor oil induced enteropooling diarrhea models or atropine (1mg/kg) for charcoal meal test. Safety study was performed using a standard acute toxicity study procedure. The effect of the extract on castor oil induced diarrheal drops, onset of diarrhea, weight of faeces, small intestinal fluid accumulation, and intestinal motility was measured and analyzed using one-way ANOVA. Preliminary phytochemical screening of the leaves powder of the plant indicated the presence of various components. Inhibition of castor oil induced diarrhea was observed at all tested doses. It can be concluded that crude extracts of Discopodium Penninervum showed strong activities against diarrhea indicating that it contains some chemical constituents that possibly lead to antidiarrheal drug development.
... A polyphenolic fraction of the aqueous extract of budding guava leaves exhibited strong antiangiogenic and anti-migratory activities against DU 145 human prostate cancer cells through the inhibition of the expression of the angiogenic stimulators including VEGF, IL-6, IL-8, matrix metalloproteinase (MMP)-2, and MMP-9 [15]. A hot aqueous P. guajava leaf extract significantly decreased the secretion of IL-8, a potent angiogenic and inflammatory factor, in response to enteropathogenic Escherichia coli infection of the human colon adenocarcinoma (HT-29) cell line [16]. In this study, we explored the anti-angiogenic and anticancer potential of the extracts of the P. ...
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Objective: To evaluate the anti-angiogenic and anticancer activities of Psidium guajava leaf extracts against angiogenesis-dependent colorectal cancer. Methods: Three extracts were produced using distilled water, ethanol, and n-hexane as solvents. The extracts were physically characterised through gas chromatography–mass spectrometry, ultraviolet-visible spectroscopy, and Fourier transform infrared spectroscopy. Their antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl, total phenolic content, and total flavonoid content assays. To assess their anti-angiogenic activity, cell viability and rat aortic ring assays were conducted, while cell migration, tube formation, colony formation, and VEGF ELISA assays were conducted to elucidate their effects on different aspects of angiogenesis. Molecular docking was used to assess the anti- angiogenic potential of some possible compounds in the extracts. Tumour spheroid assay was used to assess the extracts’ potential as a treatment for colorectal cancer. Results: The ethanol extract showed the best antioxidant activity. The distilled water and ethanol extracts exhibited more inhibitory activity against EA.hy926 cell viability and aortic ring microvessel growth. In addition, the ethanol extract performed significantly better than the distilled water extract against cell migration and colony formation, and VEGF expression of the cells was suppressed by the ethanol extract. Both the distilled water and ethanol extracts showed significant inhibitory effect on EA.hy926 tube formation and tumour spheroids consisting of EA.hy926 and HCT116 cells. The ethanol extract contained β-caryophyllene and β-elemene by phytochemical analysis and subsequent docking studies, which may contribute to its anti-angiogenic activity. Conclusions: The ethanol extract of Psidium guajava has potential in the treatment of colorectal cancer through the inhibition of angiogenesis.
... Studies undertaken at the FMR have revealed that guava leaf decoction (GLD) shows anti-rotaviral, anti-giardial [24] and antibacterial activity against V. cholerae and Shigella sp. [25] Although the decoction did not show bactericidal activity against E. coli, it inhibited the colonization and production of labile toxin and also inhibited the IL-8 production [25,26]. The invasion by S. flexneri into HEp-2 cells and production of cholera toxin were also affected [25]. ...
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Background Diarrhoea is amongst the first ten causes of death and its treatment faces an increased threat of drug resistance. Previous studies on the guava leaf decoction (GLD) revealed its suitability for use in infectious diarrhoea of unknown etiology. Objective The objective of this trial was to establish efficacy, dose and safety of GLD prepared from the Indian Sardar variety in adults with acute infectious diarrhoea. Methods The current trial was an open efficacy randomized 5-day, parallel group multi-arm interventional study. Amongst 137 adults (18–60 years) suffering with acute diarrhoea, 109 were included (57% females, 43% males). Three doses of GLD (6-leaf, 10-leaf and 14-leaf) were compared with controls receiving oral rehydration solution. Decrease in stool frequency and improvement in consistency were the outcomes measured. The data was analyzed using ANOVA, Tukey's post-hoc test, Kruscal-Wallis test and Chi-Square test where applicable. Results The trial showed that the 14-leaf (7.4 g) decoction was the most effective. Administration of the decoction, thrice daily helped the patients regain normalcy in 72 h as opposed to 120 h in controls. Safety of the intervention was reflected by normal levels of haemoglobin, liver and kidney parameters. No adverse events were reported. Conclusion The 14 leaves decoction was a safe treatment for adult acute uncomplicated diarrhoea of unknown etiology. Moreover due to component synergy and divergent mechanisms of action, it could possibly combat the generation of drug resistance and destruction of gut microbiota. Hence GLD has the potential for development as a first line treatment for diarrhoea. Trial registration Trial was registered with Clinical Trials Registry - India (CTRI registration number: CTRI/2016/07/007095). The trial was retrospectively registered.
... Nigeria was estimated to have a total number of annual child deaths due to diarrhoea to be 151,700 [9]. Diarrhoea may be caused by a wide array of agents such as entero-pathogenic microorganisms (Shigella flexneri and Shigella dysenteriae, Staphylococcus aureus, Escherichia coli, Salmonella typhi and Candida albicans), alcohol, irritable bowel syndrome, bile salts, hormones, secretory tumors and intoxication [10,11]. Dependency on plants as medicine in controlling diseases is common among rural populace in Nigeria because of its relative safety and affordability compared with the cost of conventional medicines. ...
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Full-text available
Aim: To evaluate the antidiarrhoea effect of hydromethanolic leave extract of I. asarifolia (HLEIA) on castor oil-induced diarrhea. Place and Duration of Study: Department of Biochemistry, Faculty of Life sciences, Kebbi State University of Science and Technology, Aliero, Kebbi state, Nigeria. P.M.B.1144. Kebbi State. Nigeria, between February 2015 and September 2016. Methodology: In a continuous effort to search for bioactive agents from medicinal plants, the antidiarrhoea activity of I. asarifolia was investigated. The effect of hydromethanolic leave extract of I. asarifolia (HLEIA) on castor oil-induced diarrhoea, gastrointestinal transit and intestinal fluid accumulation (enteropooling) were assessed in albino rats. Qualitative phytochemical analysis was carried out using standard procedures while acute oral toxicity studies was determined using the staircase method. Results: The phytochemical analysis showed the presence of alkaloid, terpenoid, tannin, saponin, phenols. The LD50 was estimated to be greater than 3000 mg/kg since there was no mortality recorded after 14 days of acute oral toxicity studies. Sub-chronic administration of graded doses (150 – 600 mg/kg) of HLEIA significantly (p<0.05) reduced diarrhoea episodes, decreased gastro intestinal movement and inhibited intestinal fluid accumulation compared to the control. The antidiarrhoea effect of treated group (600 mg/kg ) was comparable to that of the standard drug Loperamide. Conclusion: The findings of the present study scientifically validate the use of I. asarifolia in the treatment of diarrhoea.
... Campylobacter species, S. aureus, and C. difficile), and parasitic (Cryptosporidium and Giardia) agents are important pathogens, although rotavirus is the major cause of infectious diarrhea, particularly among young children [4][5][6][7]. ...
Article
Full-text available
Diarrhea is amajor health problem throughout theworld and it has becomemore problematic in developing countries like Ethiopia. People, in several parts of the world, use different traditional medicines for treating diarrhea and it has been reported that the roots, leaves, and flowers of various species are used for the same purpose. In Ethiopia, for instance, Discopodium Penninervum is used for the treatment of diarrhea and also to control infection. The aim of the present study was, therefore, to evaluate the in vivo antidiarrheal and in vitro antimicrobial effect of Discopodium Penninervum in mice. For the antimicrobial activity test, four standard bacteria and disc diffusion method were used, while for antidiarrheal experiment, animals had been used, which were divided into 5 groups. The first group served as negative control and was administered with vehicle (0.2-0.3ml of distilled water). Groups two (D100), three (D200), and four (D400) were administered 100, 200, and 400 mg/kg of the extract, respectively. Group five served as positive control group and was administered with either loperamide (3mg/kg) for castor oil induced diarrhea and castor oil induced enteropooling diarrhea models or atropine (1mg/kg) for charcoal meal test. Safety study was performed using a standard acute toxicity study procedure. The effect of the extract on castor oil induced diarrheal drops, onset of diarrhea, weight of faeces, small intestinal fluid accumulation, and intestinal motility wasmeasured and analyzed using one-way ANOVA. Preliminary phytochemical screening of the leaves powder of the plant indicated the presence of various components. Inhibition of castor oil induced diarrhea was observed at all tested doses. It can be concluded that crude extracts of Discopodium Penninervum showed strong activities against diarrhea indicating that it contains some chemical constituents that possibly lead to antidiarrheal drug development.
... Campylobacter species, S. aureus, and C. difficile), and parasitic (Cryptosporidium and Giardia) agents are important pathogens, although rotavirus is the major cause of infectious diarrhea, particularly among young children [4][5][6][7]. ...
Article
Full-text available
Diarrhea is a major health problem throughout the world and it has become more problematic in developing countries like Ethiopia. People, in several parts of the world, use different traditional medicines for treating diarrhea and it has been reported that the roots, leaves, and flowers of various species are used for the same purpose. In Ethiopia, for instance, Discopodium Penninervum is used for the treatment of diarrhea and also to control infection. The aim of the present study was, therefore, to evaluate the in vivo antidiarrheal and in vitro antimicrobial effect of Discopodium Penninervum in mice. For the antimicrobial activity test, four standard bacteria and disc diffusion method were used, while for antidiarrheal experiment, animals had been used, which were divided into 5 groups. The first group served as negative control and was administered with vehicle (0.2-0.3ml of distilled water). Groups two (D100), three (D200), and four (D400) were administered 100, 200, and 400 mg/kg of the extract, respectively. Group five served as positive control group and was administered with either loperamide (3mg/kg) for castor oil induced diarrhea and castor oil induced enteropooling diarrhea models or atropine (1mg/kg) for charcoal meal test. Safety study was performed using a standard acute toxicity study procedure. The effect of the extract on castor oil induced diarrheal drops, onset of diarrhea, weight of faeces, small intestinal fluid accumulation, and intestinal motility was measured and analyzed using one-way ANOVA. Preliminary phytochemical screening of the leaves powder of the plant indicated the presence of various components. Inhibition of castor oil induced diarrhea was observed at all tested doses. It can be concluded that crude extracts of Discopodium Penninervum showed strong activities against diarrhea indicating that it contains some chemical constituents that possibly lead to antidiarrheal drug development.
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We evaluated the plants used to treat inflammatory diseases by Dayak Uud Danum by examining the anti-allergic activity and the preventive effects on the damage of colon epithelial cells. The plant species examined are Callicarpa longifolia Lam, Myrmeconauclea strigosa Merr., Tebelion Eusideroxylon zwageri Teijsm & Binn., Scoparia dulcis L. and Lagerstroemia speciosa (L.) Pers. Anti-inflammatory activities of methanol extracts from leaves of these plants were analyzed in terms of delayed-type hypersensitivity against 2,4,6-trinitrochlorobenzene using BALB/cAJc mice and in vitro model of intestinal inflammation using FPCK-1-1 human colon epithelial cells. The yield percentage of methanol extract ranges from 4.33 to 8.99%. Extracts from C. longifolia, M. strigosa, E. zwageri, and S. dulcis have suppressed delayed-type hypersensitivity at both 24 and 48 hours after challenge. These extracts also suppressed the migration of eosinophils to the site of inflammation. L. speciosa extract suppressed the delayed-type hypersensitivity only at 48 hours after challenge. M. strigosa extract showed the preventive effect in the damage model of colon epithelial cells by inhibiting the decrease of transepithelial electrical resistance. Methanol extract of M. strigosa stimulated FPCK1-1 cells to produce polysaccharides and that of C. longifolia induced FPCK1-1 cells to produce IL-22. The production of polysaccharides by FPCK-1-1 cells may explain, in part, the preventive effects of the plant extracts on the damage of human colon epithelial cells. It is suggested that leaves from five medicinal plants used by Dayak Uud Danum have the potential as anti-inflammatory agents.
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The antidiarrheal effects of the aqueous leaf extract of C. sesamoides at 25, 50 and 100 mg/kg body weight was evaluated in female rats using gastrointestinal transit, diarrhea and enteropooling induced by castor oil models. The extract was positive for alkaloids, saponins, flavonoids and phenolics. The 25 mg/kg body weight of the extract significantly (p<0.05) prolonged the onset time of diarrhea, decreased the fecal parameters (number, water content, fresh weight, total number of wet feaces) with no episode in the animals treated with 50 and 100 mg/kg body weight. The activity of small intestine Na+-K+ ATPase increased (p<0.05) while the nitric oxide, volume and mass of intestinal fluid as well as the distance travelled by the charcoal meal decreased. The patterns of changes were similar to the reference drugs. Overall, the antidiarrheal activity of the aqueous leaf extract of Ceratotheca sesamoides may be due to alkaloids, phenolics, flavonoids and saponins present in the extract.
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Zingiber officinale (ginger) was studied for its antimicrobial profile and effect on virulent features of diarrhoeal pathogens, viz. colonization of epithelial cells and production of enterotoxins. Z. officinale showed no antimicrobial activity. Although it inhibited the production of cholera toxin, it had no effect on the action of this toxin. It also had no effect on the production and action of E. coli heat labile and heat stable toxins. However the bacterial colonization of HEp-2 cells was reduced. The results indicate that in the absence of antimicrobial action, Z. officinale exhibits its antidiarrhoeal activity by affecting bacterial and host cell metabolism. The present study reports a novel mechanism of action by Z. officinale in infectious diarrhoea.
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The aqueous extract of Psidium guajava budding leaves (PE) bears an extremely high content of polyphenolic and isoflavonoids. Whether it could be used as an anti-tumor chemopreventive in view of anti-angiogenesis and anti-migration, we performed the assay methods including the MTT assay to examine the cell viability; the ELISA assay to test the expressions of VEGF, IL-6 and IL-8; the western blot analysis to detect TIMP-2; the gelatinolytic zymography to follow the expression of MMPs; the wound scratch assay to examine the migration capability; and the chicken chorioallantoic membrane assay to detect the suppressive angiogenesis. Results indicated that the IC50 of PE for DU145 cells was ∼0.57 mg ml −1 . In addition, PE effectively inhibited the expressions of VEGF, IL-6 and IL-8 cytokines, and MMP-2 and MMP-9, and simultaneously activated TIMP-2 and suppressed the cell migration and the angiogenesis. Conclusively, PE potentially possesses a strong anti-DU145 effect. Thus, clinically it owns the potential to be used as an effective adjuvant anti-cancer chemopreventive.
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In an attempt to scientifically appraise the ‘ healing powers\' and medicinal value of Harpagophytum procumbens DC root aqueous extract (HPE), and throw some light on the efficacy and safety of the medicinal plant product, the cardiovascular effects of the herb\'s root aqueous extract (HPE) have been investigated in some mammalian experimental animal models. The results of this laboratory animal study indicate that relatively low to moderate doses of H. procumbens root aqueous extract (HPE, 10–400 mg/kg i. v.) produced dose-dependent hypotensive and cardio-depressant effects on systemic arterial blood pressures and heart rates of pentobarbitone-anaesthetized rats. Relatively low to high concentrations of the plant\'s extract (HPE, 10–1000 g/ml) also produced concentration-related biphasic responses in isolated cardiac muscle strips of guinea-pigs and isolated portal veins of rats. Relatively low concentrations of the plant\'s extract (HPE, 10–100 g/ml) always produced initial slight, transient and non-significant (P 0.05) positive chronotropic responses in isolated spontaneouslybeating right atria, but significant (P 0.05) positive inotropic responses in isolated electricallydriven left atria of guinea-pigs. However, moderate to high concentrations of the plant\'s extract (HPE, 400–1000 g/ml) always induced dose-dependent, significant (P 0.05–0.001), secondary longer-lasting, negative chronotropic and inotropic responses of the isolated spontaneouslybeating right-, and isolated electrically-driven left-, atrial muscle preparations of guinea-pigs. The plant\'s extract also produced concentration-related biphasic effects on rat isolated portal vein. Low to high concentrations of the plant\'s extract (HPE, 10–1000 g/ml) always produced dose-dependent, initial slight, transient and significant (P 0.05–0.001) contractions of the rat isolated portal veins, followed by secondary, longer-lasting, significant (P 0.05–0.001) relaxations of the muscle preparations. Although the precise mechanisms of the hypotensive and cardio-depressant actions of HPE are unknown, the vasorelaxant action of the plant\'s extract is speculated to contribute, at least in part, to the hypotensive action of the plant\'s extract. The results of this laboratory animal study lend pharmacological credence to the suggested folkloric uses of Harpagophytum procumbens secondary root in the management and/or control of hypertension and certain cardiac disorders in some communities of South Africa. Key Words: Harpagophytum procumbens, cardiovascular effects. Afr. J. Trad. CAM (2004) 1: 30-44
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There is persuasive epidemiological and experimental evidence that dietary polyphenols have anti-inflammatory activity. Aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) have long been used to combat inflammation. Recently, cyclooxygenase (COX) inhibitors have been developed and recommended for treatment of rheumatoid arthritis (RA) and osteoarthritis (OA). However, two COX inhibitors have been withdrawn from the market due to unexpected side effects. Because conventional therapeutic and surgical approaches have not been able to fully control the incidence and outcome of many inflammatory diseases, there is an urgent need to find safer compounds and to develop mechanism-based approaches for the management of these diseases. Polyphenols are found in many dietary plant products, including fruits, vegetables, beverages, herbs, and spices. Several of these compounds have been found to inhibit the inflammation process as well as tumorigenesis in experimental animals; they can also exhibit potent biological properties. In addition, epidemiological studies have indicated that populations who consume foods rich in specific polyphenols have lower incidences of inflammatory disease. This paper provides an overview of the research approaches that can be used to unravel the biology and health effects of polyphenols. Polyphenols have diverse biological effects, however, this review will focus on some of the pivotal molecular targets that directly affect the inflammation process.
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Twelve compounds were isolated from the MeOH extract of the leaf and stem of the Korean endemic plant Weigela subsessilis L. H. Bailey. Their chemical structures were elucidated on the basis of physicochemical and spectroscopic data and by comparison with those of published literatures. These compounds were identified as three sterols, P-sitosterol acetate (2), beta-sitosterol (3), daucosterol (11), eight triterpenoids, squalene (1), ursolic acid (4), ilekudinol A (5), corosolic acid (6), ilekudinol B (7), esculentic acid (8), pomolic acid (9), asiatic acid (10), and one iridoid glycoside, alboside 1 (12). This is the first report pertaining to the isolation of these compounds from Weigela subsessilis L. H. Bailey. In addition, three compounds 7, 9, and 12 were found to display a strong inhibitory effect on the production of IL-8 in the HT29 cells stimulated by TNF-alpha.
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A new pimarane-type diterpene compound, acanthokoreoic acid A together with three known compounds, acanthoic acid, acanthol, and sumogaside were isolated from a CH2CI2 fraction of Acanthopanax koreanum by repeated column chromatography and reversed phase preparative HPLC.,Acanthoic acid was isolated in high yields and showed potent inhibitory activity on the IL-8 secretion of the TNF-alpha-stimulated human colon adenocarcinoma cell line HT-29 and on the TNF-alpha secretion of the trypsin-stimulated human leukemic mast cell line HMC-1.
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Escherichia coli is the predominant nonpathogenic facultative flora of the human intestine. Some E. coli strains, however, have developed the ability to cause disease of the gastrointestinal, urinary, or central nervous system in even the most robust human hosts. Diarrheagenic strains of E. coli can be divided into at least six different categories with corresponding distinct pathogenic schemes. Taken together, these organisms probably represent the most common cause of pediatric diarrhea worldwide. Several distinct clinical syndromes accompany infection with diarrheagenic E. coli categories, including traveler's diarrhea (enterotoxigenic E. coli), hemorrhagic colitis and hemolytic-uremic syndrome (enterohemorrhagic E. coli), persistent diarrhea (enteroaggregative E. coli), and watery diarrhea of infants (entero-pathogenic E. coli). This review discusses the current level of understanding of the pathogenesis of the diarrheagenic E. coli strains and describes how their pathogenic schemes underlie the clinical manifestations, diagnostic approach, and epidemiologic investigation of these important pathogens.
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The aim of this study was to investigate whether treatment with a ginger (Zingiber officinale) extract of high-fat diet (HFD)-fed rats suppresses Nuclear factor-kappa B (NF-κB)-driven hepatic inflammation and to subsequently explore the molecular mechanisms in vitro. Adult male Sprague-Dawley rats were treated with an ethanolic extract of Zingiber officinale (400 mg/kg) along with a HFD for 6 weeks. Hepatic cytokine mRNA levels, cytokine protein levels and NF-κB activation were measured by real-time PCR, Western blot and an NF-κB nuclear translocation assay, respectively. In vitro, cell culture studies were carried out in human hepatocyte (HuH-7) cells by treatment with Zingiber officinale (100 μg/mL) for 24 hr prior to interleukin-1β (IL-1β, 8 ng/mL)-induced inflammation. We showed that Zingiber officinale treatment decreased cytokine gene TNFα and IL-6 expression in HFD-fed rats, which was associated with suppression of NF-κB activation. In vitro, Zingiber officinale treatment decreased NF-κB-target inflammatory gene expression of IL-6, IL-8 and serum amyloid A1 (SAA1), while it suppressed NF-κB activity, IκBα degradation and IκB kinase (IKK) activity. In conclusion, Zingiber officinale suppressed markers of hepatic inflammation in HFD-fed rats, as demonstrated by decreased hepatic cytokine gene expression and decreased NF-κB activation. The study demonstrates that the anti-inflammatory effect of Zingiber officinale occurs at least in part through the NF-κB signalling pathway.