In vitro susceptibility of Helicobacter pylori to isoquinoline alkaloids from Sanguinaria canadensis and Hydrastis canadensis


Mahady, G.B.; Pendland, S.L.; Stoia, A.; Chadwick, L.R.

PhytoTherapy Research 17(3): 217-221

2003


Methanol extracts of the rhizomes of Sanguinaria canadensis, and the roots and rhizomes of Hydrastis canadensis, two plants used traditionally for the treatment of gastrointestinal ailments, were screened for in vitro antibacterial activity against 15 strains of Helicobacter pylori. The rhizome extracts, as well as a methanol extract of S. canadensis suspension-cell cultures inhibited the growth of H. pylori in vitro, with a MIC50 range of 12.5-50.0 micro g/ml. Three isoquinoline alkaloids were identified in the active fraction. Sanguinarine and chelerythrine, two benzophenanthridine alkaloids, inhibited the growth of the bacterium, with an MIC50 of 50.0 and 100.0 micro g/ml, respectively. Protopine, a protopine alkaloid, also inhibited the growth of the bacterium, with a MIC50 of 100 micro g/ml. The crude methanol extract of H. canadensis rhizomes was very active, with an MIC50 of 12.5 micro g/ml. Two isoquinoline alkaloids, berberine and beta -hydrastine, were identified as the active constituents, and having an MIC50 of 12.5 and 100.0 micro g/ml, respectively.

PHYTOTHERAPY
RESEARCH
Phytother.
Res.
17,
217-221
(2003)
Published
online
in
Wiley
InterScience
(www.interscience.wiley.com
).
DOI:
10.1002/ptr.1108
In
Vitro
Susceptibility
of
Helicobacter
pylori
to
Isoquinoline
Alkaloids
from
Sanguinaria
canadensis
and
Hydrastis
canadensis
Gail
B.
Mahady,
1
'
2
Susan
L.
Pendland,
2
Adenia
Stoia
l
and
Lucas
R.
Chadwick
s
'Program
for
Collaborative
Research
in
the
Pharmaceutical
Sciences,
College
of
Pharmacy,
University
of
Illinois
at
Chicago,
Chicago,
Illinois
'Department
of
Pharmacy
Practice,
College
of
Pharmacy,
University
of
Illinois
at
Chicago,
Chicago,
Illinois
Methanol
extracts
of
the
rhizomes
of
Sanguinaria
canadensis,
and
the
roots
and
rhizomes
of
Hydrastis
canadensis,
two
plants
used
traditionally
for
the
treatment
of
gastrointestinal
ailments,
were
screened
for
in
vitro
antibacterial
activity
against
15
strains
of
Helicobacter
pylori.
The
rhizome
extracts,
as
well
as
a
methanol
extract
of
S.
canadensis
suspension-cell
cultures
inhibited
the
growth
of
H.
pylori
in
vitro,
with
a
MIC,
range
of
12.5-50.0
µg/ml.
Three
isoquinoline
alkaloids
were
identified
in
the
active
fraction.
Sanguinarine
and
chelerythrine,
two
benzophenanthridine
alkaloids,
inhibited
the
growth
of
the
bacterium,
with
an
MIC„
of
50.0
and
100.0
µg/ml,
respectively.
Protopine,
a
protopine
alkaloid,
also
inhibited
the
growth
of
the
bacterium,
with
a
MIC,
of
100
µg/ml.
The
crude
methanol
extract
of
H.
canadensis
rhizomes
was
very
active,
with
an
MIC„
of
12.5
µg/ml.
Two
isoquinoline
alkaloids,
berberine
and
p-hydrastine,
were
identified
as
the
active
constituents,
and
having
an
MIC,
of
12.5
and
100.0
µg/ml,
respectively.
Copyright
©
2003
John
Wiley
&
Sons,
Ltd.
Keywords:
Helicobacter
pylori;
Sanguinaria
canadensis;
Hydrastis
canadensis;
berberine;
sanguinarine.
INTRODUCTION
The
discovery
of
the
bacterium
Helicobacter
pylori
(HP)
as
the
main
etiologic
organism
of
chronic
gastritis,
peptic
ulcer
disease,
and
gastric
cancer
was
the
most
significant
discovery
in
the
field
of
gastroenterology
of
the
twentieth
century
(Graham,
1989,
1994).
Helicobacter
pylori-induced
gastritis
is
now
associated
with
duodenal
ulcer
disease,
peptic
ulcer
disease,
gastric
carcinoma,
primary
gastric
B-cell
lymphoma,
ischemic
heart
disease
and
hyperemesis
gravidarum
(Goodwin,
1997;
Kuipers
and
Appelmelk,
1997;
Frigo
et
aL,
1998;
Markus
and
Mendall,
1998).
Statistics
from
1997
indicate
that
as
much
as
one-half
of
the
world's
population
is
infected
with
the
bacterium
(Breuer
et
al.,
1997).
Treatment
of
HP
infections
often
consists
of
triple
drug
therapy,
including
a
proton
pump
inhibitor
or
bismuth
salts
in
combination
with
two
antibiotics
for
7-14
days
(Sung
et
al.,
1996;
Salcedo
and
Al
Kawas,
1998).
While,
triple
and
quadruple
drug
therapy
is
usually
effective,
anti-
biotic
resistant
strains
of
H.
pylori
have
emerged,
and
are
becoming
problematic
throughout
the
world
(Graham
et
al.,
1996;
Fedorak
et
al.,
1997;
Dore
et
al.,
1999).
In
view
of
the
development
of
antibiotic
resistance,
HP
is
now
viewed
as
a
significant
human
pathogen
in
need
of
*
Correspondence
to:
Dr
G.
B.
Mahady,
PCRPS,
Room
310,
College
of
Pharmacy,
University
of
Illinois,
833
S.
Wood
St.,
MC
877,
Chicago,
Il
60612,
USA.
Tel:
312
996-1669.
Fax:
312
413-5894.
E-mail:
mahady@uic.edu
Contract/grant
sponsor:
National
Center
for
Complementary
and
Altern-
ative
Medicine;
Contract/grant
number:
At00413.
Contract/grant
sponsor:
NIH.
Contract/grant
sponsor:
Hans
Valhteich
Scholarship
award.
Copyright
©
2003
John
Wiley
&
Sons,
Ltd.
new
chemotherapeutic
agents
for
treatment
and
pre-
vention
(Doig
and
Trust,
1997).
The
discovery
and
development
of
botanical
extracts
for
the
treatment
of
HP
infections
is
one
such
strategy.
Plant-based
medicines
have
been
used
throughout
history,
in
traditional
systems
of
medicine,
to
treat
a
variety
of
gastrointestinal
ailments,
including
stomachache,
gastritis,
diarrhea,
and
peptic
ulcers.
Two
such
plants,
Sanguinaria
canadensis
L.
(bloodroot,
Papaveraceae)
and
Hydrastis
canadensis
L.
(goldenseal,
Ranunculaceae),
are
herbaceous
perennials
native
to
eastern
Canada
and
the
United
States
(Der
Marderosian,
1977;
Mahady,
1993).
Traditionally,
the
Native
American
Indians
used
hot
water
extracts
of
both
plants
for
the
treatment
of
a
variety
of
ailments
including
dyspepsia,
gastritis
and
indigestion
(Anderson,
1885;
Veninga
and
Zaricor,
1976;
Mahady
et
aL,
1993).
The
rhizomes
of
S.
canadensis
contain
an
acrid
orange-red
juice
from
which
numerous
isoquinoline
alkaloids,
including
chelerythrine,
protopine and
sanguinarine
(Fig.
1)
have
been
isolated
(Mahady
et
al.,
1993).
Experimental
evidence
has
shown
that
aqueous
and
alcohol
extracts
of
the
rhizomes
inhibit
the
growth
of
Staphylococcus
aureus
and
Mycobacterium
tubercu-
losis
in
vitro,
and
the
constituents
responsible
for
this
activity
were
two
of
the
benzophenanthridine
alka-
loids,
sanguinarine
and
chelerythrine
(D'Amico,
1950;
Gottshall
et
al.,
1949).
Similarly,
the
roots
and
rhizomes
of
H.
canadensis
produce
a
yellow-orange
juice
con-
taining
isoquinoline
alkaloids,
including
berberine
and
/3-hydrastine
(Fig.
1)
(Der
Marderosian,
1977)
Crude
extracts
of
the
rhizome,
as
well
as
berberine
have
been
shown
to
inhibit
the
growth
of
Staphylococcus
aureus
and
E.
coli
in
vitro
(D'Amico,
1950;
Hocking,
1977).
Received
1
October
2001
Revised
15
November
2001
Accepted
22
January
2002
218
G.
B.
MAHADY
ET
AL.
OMe
OMe
O
\-0
Sanguinarine
Berberine
MeO
MeO
Chelerythrine
Protopine
O
Fe'
OMe
OMe
fi-Hydrastine
Figure
1.
Because
of
their
traditional
use
for
the
treatment
of
gastrointestinal
disorders,
and
their
purported
anti-
bacterial
activities,
methanol
extracts
of
both
plants
were
screened
against
15
strains
of
H.
pylori.
This
report
describes
the
results
of
the
in
vitro
susceptibility
of
H.
pylori
to
the
methanol
extracts,
as
well
as
their
constituent
isoquinoline
alkaloids.
MATERIALS
AND
METHODS
Plant
Materials
and
Extraction
The
cultivated
rhizomes
of
Sanguinaria
canadensis
L.
were
collected,
cleaned
and
air-dried
before
grinding.
The
ground
rhizomes
(100
g)
were
extracted
three
times
with
500
ml
of
95%
methanol,
and
evaporated
to
dryness
under
reduced
pressure.
The
dried
crude
methanol
extracts
were
re-dissolved
in
a
minimal
amount
of
methanol
and
eluted
through
a
Prep
Sep®
cyano
column
to
produce
extracts
with
a
high
concen-
tration
of
isoquinoline
alkaloids
(Mahady
et
al.,
1993).
The
alkaloids,
sanguinarine
and
chelerythrine,
and
protopine
were
isolated
and
characterized
from
the
concentrated
extracts
using
a
previously
described
protocol
(Mahady
and
Beecher,
1994).
Suspension
cell
cultures
of
S.
canadensis
were
maintained
on
a
modified
Gamborg
B5
medium
(2
g
of
tissue
per
100
ml
media),
and
subcultured
every
two
weeks
(Mahady
et
al.,
1993).
The
suspension
cells
(100
g)
were
harvested
at
fourteen
days,
dried
and
9.86
g
were
extracted
three
times
with
100
ml
of
95%
methanol.
The
extract
was
dried
under
reduced pressure
and
then
re-dissolved
in
a
minimal
amount
of
methanol
and
eluted
through
a
Prep
Sep®
cyano
column
as
previously
described
above.
Hydrastis
canadensis
dried
powdered
roots
and
rhizomes
were
obtained
from
Frontier
Natural
Products,
Norway,
Iowa.
The
powdered
plant
materials
(1718
g)
were
extracted
with
95%
methanol
(MeOH),
three
times,
4000
ml
for
48
h,
3500
ml
for
24
h
and
3300
ml
for
24
h.
The
Me0H
extract
was
evaporated
under
reduced
pressure
leaving
a
dark
brown
residue
which
was
dissolved
in
100%
Me0H
and
filtered,
resulting
in
a
yellowish-brown
crystalline
substance.
Repeated
recrystal-
lization
from
chloroform/MeOH
yielded
pure
/3-
hydrastine
(colorless
crystals)
and
berberine
(yellow
crystals).
The
extracts
and
pure
compounds
were
stored
Copyright
©
2003
John
Wiley
&
Sons,
Ltd.
Phytother.
Res.
17,
217-221
(2003)
IN
VITRO
SUSCEPTIBILITY
OF
HELICOBACTER
PYLORI
219
at
-20
°C
in
sterile
borosilicate
glass
vials.
A
10
mg
sample
of
each
plant
extract
or
pure
compound
was
tested
for
antibacterial
activity
using
the
HP
in
vitro
susceptibility
assay.
H.
pylori
assay
Susceptibility
testing
was
performed
using
the
agar
dilu-
tion
procedure
according
to
the
guidelines
described
by
the
National
Committee
for
Clinical
Laboratory
Standards
(1997,
1999).
The
extracts
were
dissolved
in
methanol
and
sterile
distilled
water
was
used
for
further
serial
dilutions
of
the
dissolved
plant
extracts.
Final
test
concentrations
consisted
of
100,
50,
25,
and
12.5,
6.25,
3.125,
1.56,
0.78
µg/m1
for
each
sample.
One
mL
of
each
concentration
was
added
to
19
mL
of
molten
Mueller-
Hinton
agar
(pH
7.3)
supplemented
with
10%
sterile
defibrinated
horse
blood.
Growth
control
plates
consist-
ing
of
20
mL
of
agar
medium
were
included
in
each
experiment.
Petri
plates
incorporating
minimal
to
maximum
volumes
of
vehicle
solvent
were
included
as
a
growth
control
to
ensure
the
viability
of
the
organisms
was
not
affected
by
the
solvent
used
to
dissolve
the
plant
extracts.
For
quality
control
and
comparative
analyses,
the
antibiotic
amoxicillin
was
also
tested
with
each
batch
of
plant
extracts.
A
total
of
14
clinical
isolates
namely,
assession
numbers:
A2,
A6,
Ed,
002,
019A,
1022,
1050,
1058,
1060, 1080, 1153,
1175, 1452,
4126,
and
1
American
Type
Culture
Collection
(ATCC)
(Rockville,
MD)
strain
(#43504)
of
H.
pylori
were
used
in
the
susceptibility
testing.
The
clinical
strains
were
coded
to
protect
the
identity
of
the
patient
from
which
they
were
obtained.
Some
of
the
isolates
were
obtained
from
the
Micro-
biology
Laboratory
at
the
University
of
Illinois
Medical
Center
(Chicago,
IL),
Abbott
Laboratories
(Abbott
Park,
IL),
and
Dr
D.
Y.
Graham
(Houston,
TX).
The
isolates
obtained
from
Abbott
Laboratories
include
organisms
obtained
from
patients
in
Richmond,
VA;
Charlottesville,
VA;
Nashville,
TN;
and
Southampton,
England.
Clinical
isolates
were
obtained
from
differ-
ent
geographic
regions
to
ensure
that
the
organisms
were
genetically
distinct.
Gram
stain
appearance,
and
a
positive
urease
test
confirmed
the
identification
of
each
organism.
The
organisms
were
stored
frozen
at
-70
°C
in
skimmed
milk
plus
17%
glycerol.
For
susceptibility
testing,
the
organisms
were
inocu-
lated
onto
5%
sheep
blood
agar
plates,
and
incubated
at
37°C
in
a
10%
CO
2
atmosphere
for
72
h.
The
organ-
isms
were
then
subcultured
once
to
ensure
reliable
growth.
An
inoculum
of
each
isolate
was
prepared
by
suspending
the
organism
in
4.5
ml
of
sterile
Mueller-
Hinton
broth
and
adjusting
the
turbidity
to
that
of
a
2.0
McFarland
Standard
using
a
spectrophotometer
at
625
nm.
This
density
produces
a
suspension
of
approx-
imately
1
x
10
8
CFU/ml
of
H.
pylori.
The
organisms
were
inoculated
onto
the
agar
plates
containing
con-
secutive
dilutions
of
the
plant
extracts
via
a
32-prong
inoculating
device.
The
device
delivers
8
µ1
per
spot
resulting
in
a
final
inoculum
of
approximately
1
x
10
6
CFU/spot.
After
the
spots
dried,
the
plates
were
incubated
at
37
°C
in
10%
CO
2
and
examined
for
growth
after
3
days.
All
procedures
were
performed
in
duplicate.
The
minimum
inhibitory
concentration
(MIC),
defined
as
the
lowest
concentration
of
the
com-
pound
at
which
there
was
no
visible
growth
or
only
a
faint
haze,
was
determined
for
each
plant
extract
and
pure
compound.
RESULTS
The
minimum
inhibitory
concentrations
(MIC)
of
crude
methanol
extracts
of
Sanguinaria
canadensis
and
Hydrastis
canadensis,
and
isolated
chemical
constituents
are
presented
in
Table
1.
The
95%
methanol
extracts
of
both
plants
inhibited
the
growth
of
HP
in
vitro.
The
crude
methanol
extract
of
the
roots
and
rhizomes
of
H.
canadensis
was
slightly
more
active
(MIC
50
12.5
µg/mg;
range
0.78-25
µg/m1)
than
the
crude
extract
from
the
rhizomes
or
cell
suspension
cultures
of
S.
canadensis
(MIC
50
12.5
µg/m1;
range
12.5-100
µg/m1).
Three
iso-
quinoline
alkaloids
isolated
from
the
rhizome
extract
of
S.
canadensis
namely,
sanguinarine,
chelerythrine
and
protopine,
inhibited
the
growth
of
the
bacterium
with
an
MIC
50
of
50.0
to
100.0
µg/ml.
Sanguinarine,
was
the
most
active
of
the
three
alkaloids,
and
inhibited
the
growth
of
all
15
HP
strains
with
an
MIC
range
of
6.25
to
50.0
µg
/ml
(Table
2).
Two
isoquinoline
alkaloids,
berberine
and
hydrastine,
isolated
from
H.
canadensis,
inhibited
the
growth
of
H.
pylori
with
an
MIC
50
of
12.5
and
100.0
µg/ml,
respectively.
Berberine
was
the
most
active
alkaloid
in
the
assay,
and
inhibited
the
growth
of
all
15
HP
strains,
with
an
MIC
range
of
0.78
to
25.0
µg
/ml
(Table
2).
The
MIC
of
amoxicillin
was
in
the
acceptable
range
established
by
NCCLS
against
the
control
strain,
H.
pylori
ATCC
43504
(Table
1).
Table
1.
MICs
of
Sanguinaria
canadensis
extracts
and
constituents
against
H.
pylori
Sample Sample
description
MIC,
µg/m1
MIC,
µg/m1
MIC
range
µg/m1
1.
Sanguinaria
canadensis
rhizome-MeOH
12.5
50
12.5-50
2.
S.
canadensis
Me0H-Cyano
fraction
25
50
12.5-100
3.
S.
canadensis-suspension
cell
cultures
50
50
25-50
4.
Sanguinarine
50
50
6.25-50
5.
Chelerythrine
100 100
25-100
6.
Protopine
100
>100
25-
>
100
7.
Hydrastis
canadensis
rhizome-MeOH
12.5
50
0.78-50
8.
Berberine
12.5
25
0.78-25
9.
ig-Hydrastine
100 100
25-
>
100
10.
Amoxicillin
0.002
0.06
0.002-0.06
Copyright
©
2003
John
Wiley
&
Sons,
Ltd.
Phytother.
Res.
17,
217-221
(2003)
220
G.
B.
MAHADY
ET
AL.
Table
2.
Susceptible
strains
of
HP
and
minimum
inhibitory
concentrations
of
crude
drug
extracts
and
active
isoquinoline
alkaloids.
The
clinical
HP
strains
are
list
by
assession
numbers
designated
by
the
individual
laboratories
from
where
they
originated.
They
are
coded,
either
alphabetical,
alpha-numeric,
or
numerically
Extract
or
Compound
MIC
0.7814/m1
MIC
6.2514/m1
MIC
12.514/m1
MIC
25.014/m1
MIC
50.014/m1
Tested
Sanguinaria
root
extract
ATCC
43504,
Ed,
002,
019A,
1175,
A2,
1058,
1153
1009,
1022,
1050,
A6
1060,
1080,
1452,
4126,
Sanguinarine
019A,
1022,
1050,
A2,
A6,
ATCC
43504,
1175,
1452
Ed,
1058, 1060, 1080,
1153,
4126,
002
Hydrastis
root
and
rhizome
extract
1050
1022,
1452
002,
019A,
1058,
1080,
A2
Ed,
A6,
1153,
4126
ATCC
43504,
1060
Berberine
1050
1022,
1058,
1080,
002,
A2, A6,
019A,
Ed,
4126,
1060,
1153,
1452
1175
ATCC
43504
DISCUSSION
Previously
published
investigations
have
demonstrated
that
medicinal
plants,
used
traditionally
to
treat
peptic
ulcer
disease
and
other
gastrointestinal
ailments,
also
inhibit
the
growth
of
HP
in
vitro.
For
example,
Termi-
nalia
spinosa,
an
East
African
plant,
inhibited
the
growth
of
HP,
with
a
MIC
range
of
62.5-500
µg/ml
(Fabry
et
al.,
1996).
Extracts
of
Thymus
vulgaris
and
Cinnamonum
zeylanicum
also
inhibited
the
growth
of
the
bacterium,
(Tabak
et
al.,
1996)
and
extracts
of
garlic
(Allium
sativum)
were
effective,
with
an
MIC
range
of
20-40.0
µg/ml
(Sivam
et
al.,
1997;
Jonkers,
1999).
Furthermore,
extracts
of
Pistacia
lentiscus
and
Cetraria
islandica,
plants
used
traditionally
for
the
treatment
of
dyspepsia,
also
inhibit
the
growth
of
H.
pylori
in
vitro
(Ingsdorfer
et
al.,
1997;
Huwez,
1998).
Thus,
there
is
a
growing
body
of
evidence
suggesting
that
medicinal
plants
are
good
candidates
for
developing
new
agents
for
the
treatment
and
prevention
of
HP
infections.
Traditionally,
extracts
of
S.
canadensis
and
H.
canadensis
were
employed
as
homeopathic
and
folk
remedies
for
the
treatment
of
various
illnesses,
including
dyspepsia,
gastritis
and
ulcers
(Anderson,
1885;
Veninga
and
Zaricor,
1976).
In
fact
in
1830,
tinctures
and
extracts
of
S.
canadensis
were
listed
as
digestants,
in
both
the
United
States
Pharmacopoeia
and
National
Formulary
(Harkrader,
1990).
Furthermore,
descriptions
of
the
clinical
use
of
S.
canadensis
for
the
treatment
of
gastritis
and
stomach
ulcers
were
published
as
far
back
as
1882
(Winterburn,
1885).
More
recently,
both
plants
have
been
popularized
as
ingredients
of
dietary
supplements
in
the
United
States
with
a
variety
of
structure-function
claims
(Mahady
and
Chadwick,
2001).
While
crude
extracts
of
S.
canadensis,
and
H.
canadensis
have
been
shown
to
inhibit
the
growth
of
various
microorganisms
in
vitro
(Harkrader,
1990;
Sca
z7occhio
et
al.,
1998;
Veninga
and
Zaricor,
1976;
Walker,
1990),
their
antibacterial
activity
against
H.
pylori
had
not
been
previously
reported.
In
this
investigation,
95%
methanol
extracts
of
the
roots
and/or
rhizomes
of
S.
canadensis
and
H.
canadensis
inhibited
the
growth
of
14
clinical
isolates
and
one
ATCC
strain
of
H.
pylori.
All
of
the
anti-helicobacter
activity
appeared
in
the
alkaloid
fractions
of
the
extracts,
with
the
active
con-
stituents
consisting
primarily
of
known
isoquinoline
alkaloids.
Berberine
and
sanguinarine
were
the
most
active
isoquinoline
alkaloids
tested,
and
have
previously
been
reported
to
have
antimicrobial
activities
(Walker,
1990;
Gentry
et
al.,
1998).
Berberine
has
been
reported
to
inhibit
the
growth
of
HP
in
vitro
(Bae
et
al.,
1998),
and
its
efficacy
against
HP
has
been
compared
with
that
of
other
antibiotics
in
a
clinical
trial
(Hu,
1993).
Few
side
effects
were
observed
in
patients
treated
orally
with
400
mg
of
berberine
per
day.
Although
there
is
little
in
the
way
of
in
vivo
toxicological
data
for
gold-
enseal,
there
is
some
information
is
available
for
berberine.
The
oral
median
lethal
dose
is
329
mg/kg
body
weight
in
mice
and
100
mg/kg
body
weight
in
dogs,
doses
well
above
that
needed
to
treat
HP
infec-
tions
(Lampe,
1992).
As
for
the
safety
of
sanguinarine,
no
adverse
effects
have
been
observed
in
animals
treated
with
up
to
100
mg/kg,
and
the
oral
median
lethal
dose
in
rats
was
1658
mg/kg,
thus
again
demon-
strating
low
in
vivo
toxicity
(Becci
et
al.,
1987;
Keller
and
Meyer,
1989).
The
mechanism
by
which
these
extracts
and
alkaloids
exert
their
antibacterial
effect
on
HP
is
not
known.
However,
it
has
been
shown
in
vitro
that
berberine
blocks
the
adhesion
of
uropathogenic
Escherichia
coli
to
erythrocytes
and
epithelial
cells
through
a
mechanism
that
involves
a
reduction
in
the
synthesis
of
fimbrial
subunits
and
the
expression
of
assembled
fimbriae
(Sun
et
al.,
1988).
Since,
in
vitro
and
in
vivo
studies
have
both
demonstrated
that
the
attachment
of
H.
pylori
to
the
gastric
epithelial
cells
is
mediated
by
the
fucosylated
Lewis
b
histo-blood
group
antigen-binding
adhesin
(Ilver
et
al.,
1998;
Gerhard
et
al.,
1999)
this
interesting
mechanism
of
action
will
be
explored
in
future
investiga-
tions
with
both
berberine
and
sanguinarine.
Acknowledgements
This
study
was
supported
in
part
by
Grant
No.
AT00413
from
the
National
Center
for
Complementary
and
Alternative
Medicine,
NIH,
and
the
Hans
Valhteich
Scholarship
award
to
GBM.
The
authors
also
acknowledge
the
gift
of
fl-hydrastine
by
Dr.
D.
Kinghorn,
Program
for
Collaborative
Research
in
the
Pharmaceutical
Sciences,
University
of
Illinois
at
Chicago.
Copyright
©
2003
John
Wiley
&
Sons,
Ltd.
Phytother.
Res.
17,
217-221
(2003)
IN
VITRO
SUSCEPTIBILITY
OF
HELICOBACTER
PYLORI
221
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Copyright
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2003
John
Wiley
&
Sons,
Ltd.
Phytother.
Res.
17,
217-221
(2003)