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A catechin, EGCG also demonstrated antibacterial activ-
ity. However, other compounds exhibited minimal
antibacterial activity against H. pylori with a few excep-
tions. Hydrolyzable tannins have higher water-solubil-
ity than other compounds (53). Water-solubility may be
important for their antibacterial action. Mabe et al.
have reported that tea catechins may have therapeutic
effects on H. pylori infection (25). In this study, many
hydrolyzable tannins revealed stronger antibacterial
activity than catechins. Furthermore, acid-treatment
scarcely affected the antibacterial activity of hydrolyzable
tannins in vitro, which suggests that they can work in the
acidic gastric environment. Thus, hydrolyzable tannins
may be promising therapeutic agents in the treatment of
H. pylori infection.
No compounds tested in this study demonstrated
antibacterial activity against E. coli which is a normal
inhabitant of the human intestinal tract. Hydrolyzable
tannins may have a narrow antibacterial spectrum. New
triple therapies suppress not only H. pylori but also
intestinal bacterial ora, which causes such side effects
as abdominal pain and diarrhea (46). Harsch et al. have
reported a case of pseudomembranous colitis after erad-
ication of H. pylori with a new triple therapy (9).
Hydrolyzable tannins may be able to suppress H. pylori
without affecting intestinal bacterial ora. Furthermore,
hydrolyzable tannins did not affect the viability of MKN-
28 cells derived from human gastric epithelium (28).
Thus, hydrolyzable tannins may be able to suppress H.
pylori without affecting the viability of gastric epithelial
cells.
A monomeric hydrolyzable tannin, TG-I decreased the
viability of H. pylori in time- and dose-dependent man-
ners in vitro. This result indicates that hydrolyzable
tannins have bactericidal effects against H. pylori. In the
process of bacterial killing, TG-I rapidly aggregated H.
pylori cells into clusters and fused them, which sug-
gests that hydrolyzable tannins act on the surface struc-
tures of H. pylori. Ikigai et al. have reported antibacte-
rial catechins damage bacterial membranes (13). Then,
258 K. FUNATOGAWA ET AL
Fig. 4. Membrane-damaging activity of TG-I. CF-encapsulated
liposomes were exposed to TG-I (50 g/ml). Samples were
taken at the times indicated, and the amount of CF released from
the liposomes was measured. Data are presented as means
standard deviations.
Fig. 5. Membrane-damaging activity of TG-I (), TG-II (),
acid-treated TG-I ( ), acid-treated TG-II ( ), Geraniin (),
Agrimoniin (), Oenothein A (), EGCG (), and EC (). CF-
encapsulated liposomes were exposed to each compound (3.13 to
100 g/ml). Samples were taken after 40 min of exposure, and
the amount of CF released from the liposomes was measured.
Data are presented as means standard deviations.
Table 4. Effect of plant-derived compounds on the viability of
MKN-28 cells
a)
Compound Concentration Viability (OD540)
b)
RPMI alone 0.930.06
12.5 g/ml 0.890.08
TG-I 25 g/ml 0.930.06
50 g/ml 0.940.05
12.5 g/ml 0.870.09
TG-II 25 g/ml 0.870.08
50 g/ml 0.850.09
12.5 g/ml 0.890.04
Oenothein A 25 g/ml 0.870.08
50 g/ml 0.870.08
a)
MKN-28 cells were treated with RPMI alone or containing
each compound (12.550 g/ml) for 12 hr.
b)
The viability of MKN-28 cells is expressed as the OD at 540
nm (OD540). Each value represents the mean the standard
deviation. There was no signicant difference between each
compound and RPMI alone (P0.05 by unpaired Students t test).
we investigated effects of hydrolyzable tannins and cat-
echins on lipid bilayer membranes. All hydrolyzable tan-
nins tested demonstrated dose-dependent membrane-
damaging activity. An antibacterial catechin, EGCG
showed stronger membrane-damaging activity than
hydrolyzable tannins, though it was inferior to many
hydrolyzable tannins in antibacterial activity. On the
other hand, hydrolyzable tannins did not reveal toxicity
against E. coli and MKN-28 cells.
H. pylori membrane contains three kinds of cholesteryl
glucosides (CGs); cholesteryl--D-glucopyranoside
(CGL), cholesteryl-6-O-tetradecanoyl--D-glucopyran-
oside (CAG), and cholesteryl-6-O-phosphatidyl--D-
glucopyranoside (CPG) (7, 11). These glycolipids
account for about 25% (wt/wt) of the total lipid. CGs are
found in many plants, but they are very rare in bacteria
and animals. Especially, CPG is a unique lipid found
only in Helicobacter species (8). These unique mem-
brane features might be related to the sensitivity of H.
pylori to hydrolyzable tannins.
In conclusion, plant-derived hydrolyzable tannins
have antibacterial effects against H. pylori. Hydrolyzable
tannins have potential as new and safe therapeutic regi-
mens against H. pylori infection. Whether their mem-
brane-damaging activity directly contributes to their
antibacterial action remains to be elucidated.
This study was supported in part by a Grant-in-Aid for Sci-
entic Research (No. 09672143) from the Ministry of Education,
Culture, Sports, Science and Technology, Japan.
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