ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Apr. 2009, p. 1592–1597 Vol. 53, No.

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0066-4804/09/$08.00⫹0 doi:10.1128/AAC.01242-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Antimicrobial Activity of Curcumin against Helicobacter pylori Isolates

from India and during Infections in Mice䌤
Ronita De,1 Parag Kundu,2 Snehasikta Swarnakar,2 T. Ramamurthy,1 Abhijit Chowdhury,3
G. Balakrish Nair,1 and Asish K. Mukhopadhyay1*
National Institute of Cholera and Enteric Diseases, Kolkata 700010, India1; Indian Institute of Chemical Biology, Kolkata 700032,
India2; and School of Digestive Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India3
Received 18 September 2008/Returned for modification 29 October 2008/Accepted 28 January 2009

Treatment failure is a major cause of concern for the Helicobacter pylori-related gastroduodenal diseases like
gastritis, peptic ulcer, and gastric cancer. Curcumin, diferuloylmethane from turmeric, has recently been
shown to arrest H. pylori growth. The antibacterial activity of curcumin against 65 clinical isolates of H. pylori
in vitro and during protection against H. pylori infection in vivo was examined. The MIC of curcumin ranges
from 5 ␮g/ml to 50 ␮g/ml, showing its effectiveness in inhibiting H. pylori growth in vitro irrespective of the
genetic makeup of the strains. The nucleotide sequences of the aroE genes, encoding shikimate dehydrogenase,
against which curcumin seems to act as a noncompetitive inhibitor, from H. pylori strains presenting differ-
ential curcumin MICs showed that curcumin-mediated growth inhibition of Indian H. pylori strains may not
be always dependent on the shikimate pathway. The antimicrobial effect of curcumin in H. pylori-infected
C57BL/6 mice and its efficacy in reducing the gastric damage due to infection were examined histologically.
Curcumin showed immense therapeutic potential against H. pylori infection as it was highly effective in
eradication of H. pylori from infected mice as well as in restoration of H. pylori-induced gastric damage. This
study provides novel insights into the therapeutic effect of curcumin against H. pylori infection, suggesting its
potential as an alternative therapy, and opens the way for further studies on identification of novel antimi-
crobial targets of curcumin.

Helicobacter pylori is a microaerophilic bacterium with the pylori of resistance to antibiotics, including metronidazole and
extraordinary ability to establish infections in human stomachs clarithromycin, could represent a serious problem that may
that can last for years or decades. It is carried by more than reduce treatment efficacy (10). Many studies have indicated
one-half of all people worldwide, and its prevalence exceeds that the prevalence of resistance varies geographically, ranging
90% in some developing countries like India. It has attracted from 10 to 90% for metronidazole and from 0 to 15% for
great attention as a major cause of peptic ulcer disease. In fact, clarithromycin (7, 18, 30). In view of the incomplete cure
H. pylori is the first bacterium to be classified as a group I achieved with conventional therapy because of increasingly
carcinogen by the International Agency for Research on Can- resistant strains, undesirable side effects (17), noncompliance
cer (6, 31). Based on the genetic characteristics and disease among the patients (3), the cost of the antibiotic regimens (32),
outcome, there are significant geographic differences among and a few other factors contributing to ineffectiveness, there is
H. pylori strains. Indian H. pylori strains are genetically distinct an urgent need to develop new treatment strategies for H.
from those from east Asia and the West (4, 16). Several puta- pylori infection.
tive virulence-associated factors, including alleles in the cag Previous studies have shown that the shikimate pathway is
pathogenicity island (PAI) of H. pylori, contribute to its patho- essential for the synthesis of important metabolites such as
genesis and augment the risk for gastric adenocarcinoma (14). aromatic amino acids, folic acid, and ubiquinone (20). The
As virulence markers of H. pylori are not always associated with
enzymes involved in this pathway, including shikimate dehy-
diseases, eradication of H. pylori from infected individuals re-
drogenase (SDH), have recently gained great attention as
mains the best choice for an effective treatment of H. pylori-
novel drug targets for developing antimicrobial agents that are
associated diseases. Several combination therapies have been
nontoxic (5). In support of this idea Han et al. (11) showed
formulated to eradicate this pathogen and cure or prevent
curcumin to be a noncompetitive inhibitor of SDH. This en-
associated diseases. Triple therapy, consisting of the combined
zyme, encoded by the aroE gene of H. pylori, may provide
usage of two antibiotics and a proton pump inhibitor, gives a
useful information for treating H. pylori-associated infection.
high eradication rate, producing a significant improvement in
the status of the disease (30). However, eradication by the In India since ancient times, spices and condiments have been
triple therapy is not always successful, and the acquisition by H. considered indispensable in the culinary arts, as they are used to
flavor foods. Also, these spices were recognized for their physio-
logical and medicinal properties. Curcumin (diferuloylmethane),
* Corresponding author. Mailing address: Department of Microbi- first chemically characterized in 1910, is generally regarded as
ology, National Institute of Cholera and Enteric Diseases, P 33, CIT the most active constituent of the perennial herb Curcuma
Road, Scheme XM, Beliaghata, Kolkata 700010, India. Phone: 91-33-
longa (commonly known as turmeric) (Fig. 1). Many studies
2350-1176. Fax: 91-33-2370-5066. E-mail: asish_mukhopadhyay@yahoo
.com. have attributed a wide spectrum of activities to this compound
䌤
Published ahead of print on 9 February 2009. and may provide a suitable basis for new H. pylori therapies (2,

1592
VOL. 53, 2009 EFFECT OF CURCUMIN DURING MOUSE INFECTION 1593

and adjusted to an optical density of 0.1 at 600 nm. Ten microliters of the
adjusted inoculum was delivered to BHI agar containing various concentrations
of curcumin (Sigma Chemical Co., St. Louis, MO). Curcumin was dissolved in
dimethyl sulfoxide (Sigma Chemical), and a stock concentration of 10 mg/ml was
stored at ⫺20°C. Final test concentrations consisted of 50, 40, 30, 25, 20, 15, 10,
and 5 ␮g/ml for each sample. Growth control plates containing only BHI agar
were included in each experiment. BHI agar incorporating the solvent dimethyl
sulfoxide was included as a growth control to ensure that the viability of the
organism was not affected by the solvent used to dissolve curcumin. A total of 65
clinical isolates and 1 reference strain (ATCC 43504) were used in the suscep-
tibility testing. All plates were incubated under microaerophilic conditions at
FIG. 1. Structure of curcumin. 37°C for 5 days. The MIC was defined as the lowest concentration of the
compound at which there was no visible growth. For quality control and com-
parative analyses, the antibiotic amoxicillin (Sigma Chemical Co., St. Louis, MO)
12, 26–28). A potential role for curcumin in treating Alzhei- and clarithromycin (Abbott Laboratories, Abbott Park, IL) were also tested with
each batch of curcumin.
mer’s disease and inflammatory bowel disease has been re- H. pylori infection in C57BL/6 mice. C57BL/6 mice bred in-house were used in
ported (13, 23). One study showed in vitro antimicrobial activ- all experiments. Experiments were designed to minimize animal suffering and to
ity of curcumin against H. pylori (15), but a recent study from use the minimum number associated with valid statistical evaluation, according
Italy reported that curcumin-based therapy was not effective to the guidelines of the animal ethics committee of the institute. Animals were
for eradication of H. pylori infection (8). anesthetized by ketamine (12 mg/kg of body weight), followed by cervical dislo-
cation for killing. Animals of both control and experimental groups were kept
Against this background, the present study has been con- separately in standard conditions and were fasted for 6 h with free access to water
ducted (i) to evaluate the efficacy of curcumin as an antimi- before each inoculation. Groups of mice (12 mice per group) were inoculated
crobial agent against H. pylori strains isolated from patients in with H. pylori cultures harvested in PBS twice in a period of 3 days, with about
Kolkata, India; (ii) to perform a sequence analysis of the aroE 108 CFU/mouse/inoculation (14). Mouse groups inoculated with SS1, AM1, or
PBS (control group) were kept separately, with free access to water and food.
genes, encoding SDH, the fourth enzyme involved in the shiki-
Two weeks after the final inoculation, a group of mice were orally fed with
mate pathway and a promising target for antimicrobial agents, curcumin (25 mg/kg) once daily for 7 days consecutively, while untreated infected
from strains for which curcumin MICs were different; and (iii) ones received sterile water. All mouse groups were sacrificed 3 weeks postinfec-
to understand curcumin’s effectiveness in eradicating H. pylori tion, and the gastric tissues were assessed for H. pylori colonization and histology.
infection in an animal model. DNA methods. Chromosomal DNA from mouse gastric tissues was extracted
by standard methods (24). The presence or absence of specific bacterial genes
and the specificity for the mouse genome were scored by PCR using specific
MATERIALS AND METHODS primers (Table 1) and DNA from the respective tissues. PCR was carried out in
H. pylori strains and culture. Sixty-five archived strains of H. pylori, which were 20-␮l reaction volumes using 10 pmol of each primer, 0.25 mM of each de-
isolated from antral mucosal biopsy specimens of patients at the Post Graduate oxynucleoside triphosphate, 1 U of Taq polymerase (Takara Shuzo, Japan), and
Institute of Medical Education and Research, Kolkata, India, with chronic gas- 40 ng of DNA. PCR was carried out for 40 cycles of denaturation at 94°C for 30 s,
tritis or duodenal ulcers, and 1 reference American Type Culture Collection primer-template annealing at 57°C for 30 s, and DNA synthesis at 72°C for 1 min.
strain (ATCC 43504) were used for this study. The strains were identified on the The PCR products were analyzed by electrophoresis in 2% agarose gels and
basis of colony appearance, Gram staining, and positive reactions in biochemical visualized by ethidium bromide staining. PCR product sizes were estimated with
tests (catalase, urease, and oxidase). H. pylori strains were revived and cultured 100-bp standards (New England Biolabs, MA).
on brain heart infusion (BHI) agar (Difco Laboratories, Detroit, MI) supple- To detect the changes in sequences associated with different MICs, the aroE
mented with 5% horse serum (Invitrogen, NY), 0.4% IsoVitaleX (Becton Dick- gene was amplified by PCR using primers described in Table 1. PCR fragments
inson, MD), trimethoprim (5 ␮g/ml), vancomycin (8 ␮g/ml), and polymyxin B (10 were purified for sequencing with the QIAquick PCR purification kit (Qiagen
␮g/ml). The plates were incubated at 37°C in a microaerophilic atmosphere (5% GmBH, Germany). Sequencing reactions were carried out with a BigDye Ter-
O2, 10% CO2, 85% N2) (double gas incubator; Heraeus, Langenselbold, Ger- minator cycle sequencing kit (PerkinElmer-Applied Biosystems, Foster City,
many) for 3 to 6 days. Stock cultures were maintained until use at ⫺70°C. A CA) and an automated sequencer (ABI Prism 3100 genetic analyzer; Applied
urease test was conducted with mouse gastric tissue as mentioned earlier (4). Biosystems). DNA sequence editing and analysis were performed with the
Mouse-adapted H. pylori strains. Two mouse-adapted H. pylori strains (one DNASTAR program. The aroE gene of fully sequenced strain 26695 was used
isolated from Australia [SS1] and another one from India [AM1]) were used for for comparison.
the animal experiment. They were revived and cultured for mouse infection (14). Histology. The body and the pyloric parts of stomachs from control mice and
MICs. Frozen stock cultures were streaked on BHI agar with 5% horse serum mice infected for 3 weeks and treated with curcumin were sectioned for histo-
and incubated for 3 days under microaerophilic conditions as mentioned earlier. logical studies. The tissue samples were fixed in 10% formalin and embedded in
Isolates were restreaked on fresh BHI agar and incubated for 24 h. Exponentially paraffin. The sections (5 ␮m) were cut with a microtome, stained with hematox-
growing H. pylori cells were suspended in sterile phosphate-buffered saline (PBS) ylin and eosin (29), and observed under an Olympus microscope. Images were

TABLE 1. Details of PCR primers used for analysis of vacA middle region and SDH and mouse GAPDH genes in DNA isolated from
mouse gastric tissues
Amplicon size
Locus name Primer Sequence Reference or source
(bp)

vacA m1/m2 VAG-F 5⬘-CAATCTGTCCAATCAAGCGAG-3⬘ 567/642 14

VAG-R 5⬘-GCGTCAAAATAATTCCAAGG-3⬘

aroE aroE-F 5⬘-CCAAAACGATTGGGCTGAAATTG-3⬘ 963 11

aroE-R 5⬘-AAAACGCCCTTTTCTACTAG-3⬘

GAPDH (mouse) GAPDH-F 5⬘-GCAGTGGCAAAGTGGAGATT-3⬘ 249 This study

GAPDH-R 5⬘-TCTCCATGGTGGTGAAGACA-3⬘
1594 DE ET AL. ANTIMICROB. AGENTS CHEMOTHER.

the growth of this organism. So, we focused on analyzing the

aroE gene from H. pylori strains showing different MICs. The
aroE gene of strain L1675, showing a curcumin MIC of 50
␮g/ml, was sequenced, and the sequence was compared with
the aroE gene sequence of 26695, showing a MIC of 20 ␮g/ml,
and also with those of strains PG139, PG156, and PG233,
showing MICs of 5 ␮g/ml, 30 ␮g/ml, and 5 ␮g/ml, respectively
(Fig. 3). Sequence analysis showed that the aroE genes of
L1675 and 26695 are identical and that the encoded proteins
are also identical. But analysis of the aroE genes from the other
three strains showed around 6 to 7% differences at the nucle-
otide and amino acid levels, which may reflect the polymorphic
nature of the gene. It is interesting to note that, despite the
variation observed at the nucleotide and amino acid levels for
FIG. 2. Distribution of curcumin MICs among the Helicobacter py-
strains with different curcumin MICs, the well-conserved
lori strains isolated from Kolkata, India.
amino acids, which seem to be important for substrate binding,
remain unchanged regardless of the MICs except at one posi-
tion in a single strain. This has led us to assume that, besides
captured with Camedia software (E-20P 5.0 megapixel) at original magnifications
the SDH gene, other genetic components may be involved in
of ⫻10 and ⫻20 and processed in Adobe Photoshop version 7.0.
Nucleotide sequence accession numbers. The sequences obtained here were the ability of curcumin to act as a potential nontoxigenic anti-
deposited in GenBank under accession numbers EU939304 to EU939307. microbial agent, at least with Indian strains.
Curcumin eradicates H. pylori from H. pylori-infected mouse
stomach. Since an in vitro assay showed that curcumin had
RESULTS
antibacterial effects against H. pylori, we tested the effect of
Differential inhibition of H. pylori growth in vitro by curcu- curcumin on H. pylori colonization in mice. A group of mice
min. Among 65 Helicobacter pylori strains tested against cur- were infected with AM1 and SS1 separately. Three weeks after
cumin, 47 strains were isolated from patients with duodenal the infection, they were sacrificed and gastric tissues were
ulcers, whereas 12 and 6 cases were isolated from patients with checked by a urease test, which showed positive results indi-
antral gastritis and nonulcer dyspepsia, respectively. Among cating H. pylori colonization. Establishment of H. pylori infec-
these strains, 64 strains, including ATCC 43504, showed a tion was also confirmed by PCR assay of the H. pylori-specific
positive amplicon for cagA and the other 2 provided an empty gene vacA using DNA isolated from H. pylori-infected stomach
site amplicon, indicating absence of the total cag PAI. MICs tissues. Colonization was further reconfirmed by quantitative
were determined by the agar dilution method. Curcumin in- culture in blood agar plates. Two weeks after infection, a group
hibited the growth of all 65 clinical isolates as well as the of mice were treated with curcumin for 7 days. The effect of
reference strain ATCC 43504. The MIC of curcumin ranged curcumin was assessed by a urease test with the respective
from 5 ␮g/ml to 50 ␮g/ml, and the majority of the strains (81%) mouse gastric tissues. It was observed from quantitative culture
showed a MIC of either 10 ␮g/ml (23%) or 15 ␮g/ml (58%) that curcumin, irrespective of strains, eradicated H. pylori from
(Fig. 2). A few strains showed higher MICs (7.7% and 1.5% of infected mouse stomach. To further confirm curcumin’s anti-H.
the strains yielded MICs of 30 ␮g/ml and 50 ␮g/ml, respec- pylori potential, the H. pylori-specific gene vacA was amplified
tively). These results clearly confirm that curcumin acts as a by PCR using DNA isolated from stomach tissues of H. pylori-
potent growth inhibitor for Indian H. pylori strains irrespective infected and treated mice, while the mouse-specific GAPDH
of the disease status. However, the roles of polymorphism in gene served as a control. Figure 4 shows that curcumin treat-
target genes and strain-specific differences in the MICs of ment completely eliminated H. pylori from mouse stomach.
curcumin need further investigation. The MICs of amoxicillin Thus, these in vivo data are in accordance with the in vitro
and clarithromycin ranged from 0.03 to 2 ␮g/ml and 0.02 to results, confirming curcumin’s anti-H. pylori potential.
0.12 ␮g/ml, respectively. The MICs of amoxicillin, curcumin, Histological analysis of mouse gastric tissues during H.
and clarithromycin against reference strain ATCC 43504 were pylori infection and the effect of curcumin. Groups of C57BL/6
0.03, 15, and 0.02 ␮g/ml, respectively. mice were intragastrically inoculated with SS1 strain while the
Analysis of SDH from Indian H. pylori strains for which the control group received sterile PBS. Two weeks after the final
curcumin MICs were different. The shikimate pathway is es- inoculation, a group of H. pylori-infected mice were orally fed
sential for the synthesis of important metabolites in bacteria. with 25 mg/kg curcumin for 7 days consecutively and sacrificed.
Therefore, the enzymes involved in the pathway have received Microscopic observations revealed considerable damage in the
much attention as potential drug targets for developing non- mouse gastric tissues infected with H. pylori SS1 strain for 3
toxic antimicrobial agents, herbicides, and antiparasite drugs. weeks (Fig. 5B) compared to the control (Fig. 5A). The gastric
SDH catalyzes the fourth reaction in the shikimate pathway mucosa of control tissue had an intact epithelial layer and
and is responsible for the NADPH-dependent reduction of glandular cells with continuous gastric pits (Fig. 5A). In con-
3-dehydroshikimate to shikimate. A recent study by Han et al. trast, denudation of the surface epithelial layer was clearly
(11) identified a new aroE gene encoding SDH from H. pylori, observed in the SS1-infected mouse gastric tissues; this layer
and curcumin works as a potent inhibitor of SDH. This may be was restored to almost normal after curcumin treatment.
the mechanism for how curcumin acts on H. pylori to inhibit Moreover, inflammation in the gastric pit cells, as observed in
VOL. 53, 2009 EFFECT OF CURCUMIN DURING MOUSE INFECTION 1595

FIG. 3. Amino acid sequences of SDHs encoded by aroE genes from five H. pylori strains were aligned. Sequences from L1675, PG139, PG233,
and PG156 are presented in this study. The 26695 sequence was taken from a public database. GenBank accession numbers for sequences
presented here are as follows: L1675, EU939307; PG139, EU939304; PG233, EU939305; and PG156, EU939306. Identical nucleotides are
indicated by dots. Sequence alignment was performed using the CLUSTALW program within DDBJ.

the infected tissues (Fig. 5B), was checked markedly by curcu-

min treatment (Fig. 5C). Disruption in the submucosal and
muscularis mucosal layers of the mouse gastric tissues that
occurred due to H. pylori infection (Fig. 5B) was also almost
completely abolished by curcumin treatment (Fig. 5C). Glan-
dular atrophy that occurred because of H. pylori infection (Fig.
5B) was considerably abated by curcumin therapy (Fig. 5C). In
control tissues the presence of inflammatory cells was negligi-
ble (Fig. 5D). However, infiltration of inflammatory cells was
observed in the submucosal region of H. pylori-infected mouse
gastric tissues (Fig. 5E) and was prevented to a significant
degree in curcumin-treated mice (Fig. 5F). Altogether, these
FIG. 4. Effect of curcumin on H. pylori viability in H. pylori-infected results indicate that curcumin is highly effective in healing the
mice. Lanes 1 (second from the left) and 2 represent the amplification
overall damage caused by H. pylori infection.
of the vacA middle region using DNA from the mouse-colonizing
strains AM1 and SS1 with primers VAG-F and VAG-R (Table 1).
Lane 3 represents the amplification of bacterium-specific vacA and the DISCUSSION
mouse-specific GAPDH gene using DNA isolated from the gastric
tissue of control mice. Lanes 4 and 5 represent the amplification of Eradication of H. pylori with a proton pump inhibitor-based
vacA and GAPDH using DNA isolated from the gastric tissue of H. triple therapy is presently used to treat H. pylori infection (21).
pylori-infected mice after 3 weeks of infection. Lanes 6 and 7 indi-
cate the amplification of vacA and GAPDH using DNA isolated Though it has a success rate of 80 to 90%, problems like
from the gastric tissue of curcumin-treated H. pylori-infected mice. treatment failure and contraindications for some patients are
⫹ve, positive control. common. Furthermore, rapidly emerging drug resistance in H.
1596 DE ET AL. ANTIMICROB. AGENTS CHEMOTHER.

coding SDH, from H. pylori strains showing different MICs

were analyzed for nucleotide and amino acid sequence varia-
tions. Striking differences in the nucleotide sequences of the
genes from these strains corroborated the fact that curcumin
targets the shikimate pathway and indicate that polymorphism
in aroE may lead to differential inhibition of H. pylori growth.
However, strains having identical sequences also had different
curcumin MICs. This may be due to the strain-to-strain varia-
tions in the rate of curcumin uptake/efflux, or it may be that the
antimicrobial effect of curcumin on H. pylori follows distinct
mechanisms that may not be solely dependent on inhibition of
the shikimate pathway. A recent study by Rai et al. suggests
that curcumin may inhibit bacterial cell proliferation by inhib-
iting the assembly dynamics of FtsZ (a bacterial protofila-
ment), which polymerizes to form a Z ring at the midcell that
orchestrates bacterial cell division (22).
The mouse model of H. pylori infection has been widely used
in investigations of host responses to H. pylori infection as well
as in eradication studies. In the present study, we have shown
that curcumin treatment completely eradicated H. pylori from
infected mouse stomach. This eradication by curcumin was
irrespective of the bacterial genotype, which is independent of
FIG. 5. Histology of control, H. pylori-infected, and curcumin- the presence of the cag PAI. These data are of immense im-
treated mouse gastric tissues. Histological sections were stained with portance for the development of alternative therapy against H.
hematoxylin and eosin, and photographs were taken at ⫻10 and ⫻20
pylori infection since studies on high doses of curcumin in
magnifications, respectively. (A to C) Histological appearance of gas-
tric tissues from (A) control mice, (B) mice infected with SS1 for 3 animals and humans have confirmed a lack of any toxic side
weeks, and (C) mice infected with SS1 and treated with curcumin at effects (9, 25). Histological analysis clearly showed that curcu-
10⫻ magnification. (D to F) Higher-magnification (20⫻) views of min is highly effective in repairing damaged tissue. All these
(D) control, (E) SS1-infected, and (F) SS1-infected, curcumin-treated
mouse gastric tissues. The gastric mucosal epithelium (black arrows),
observations not only indicate the therapeutic potential of cur-
gastric pits (green stars), gastric glands (green arrows), and inflamma- cumin against H. pylori infections but also highlight the anti-
tory cell infiltration (black stars) are shown. inflammatory effect of curcumin. Various clinical trials suggest
a therapeutic potential for curcumin in diseases such as famil-
ial adenomatous polyposis, inflammatory bowel disease, ulcer-
pylori strains during treatment with various antibiotics is a ative colitis, colon cancer, pancreatic cancer, hypercholester-
major obstacle for successful eradication therapies (1). Be- emia, atherosclerosis, pancreatitis, psoriasis, and arthritis (9).
cause of the prevalence of antibiotic-resistant H. pylori strains, It has been claimed that curcumin and related compounds
there is an increasing search for safe and effective nonantibi- have anti-human immunodeficiency virus type 1 and 2 activity
otic compounds that inhibit H. pylori growth. In the Indian in a recent patent application (19).
traditional medical system, a number of plants and plant prod- In conclusion, our study highlights the potential antibacterial
ucts are known to possess potent medicinal properties, sug- activity of curcumin against H. pylori in vitro, as curcumin was
gesting their usefulness in treatment. Recent studies have highly effective in inhibiting H. pylori growth irrespective of the
shown that curcumin possesses anti-H. pylori potential (11, 15).
genetic makeup of the strains, although its MIC is relatively
This prompted us to explore its antimicrobial potential against
high; this may be due to the poor bioavailability of curcumin
Indian H. pylori strains that are geographically distinct from
(22). Our studies also showed that curcumin-mediated inhibi-
east Asian and Western strains. Moreover, a majority of the
tion of H. pylori growth involved mechanisms that may not
Indian population harbors H. pylori and quite a number of
always be dependent on the shikimate pathway, which opens
them suffer from H. pylori-associated gastrointestinal diseases.
In the present study, we have primarily shown that curcumin the way for further studies directed toward determination of
potentially inhibited the growth of all the 65 H. pylori strains in novel antimicrobial targets of curcumin. Curcumin showed im-
vitro that were isolated from infected patients suffering from mense therapeutic potential against H. pylori infections and H.
gastrointestinal disorders. It is noteworthy that a majority of pylori-associated gastroduodenal diseases, as it was equally ef-
these strains were metronidazole resistant, with MICs ranging fective in eradicating H. pylori from infected mouse stomach.
from 16 ␮g/ml to ⬎64 ␮g/ml (7). So, our results suggest that Moreover, the gastric damage induced by H. pylori infection
curcumin acts through mechanisms distinctly different from was almost completely restored by curcumin, thus highlighting
the mode of action of these antibiotics for inhibition of H. its potential as an alternative therapy against H. pylori infec-
pylori growth. Han et al. (11) demonstrated that the growth- tion. Overall, this study provides novel insights in the thera-
inhibitory activity of curcumin against H. pylori is due to inhi- peutic potential of curcumin against H. pylori infections, al-
bition of the shikimate pathway, necessary for synthesis of though further studies are required to extrapolate its effect on
aromatic amino acids in bacteria. Thus, the aroE genes, en- humans.
VOL. 53, 2009 EFFECT OF CURCUMIN DURING MOUSE INFECTION 1597

ACKNOWLEDGMENTS (Curcuma longa) and curcumin inhibit the growth of Helicobacter pylori, a
group 1 carcinogen. Anticancer Res. 22:4179–4181.
The work was supported in part by the Indian Council of Medical 16. Mukhopadhyay, A. K., D. Kersulyte, J. Y. Jeong, S. Datta, Y. Ito, A.
Research, Government of India, and Program of Founding Research Chowdhury, S. Chowdhury, A. Santra, S. K. Bhattacharya, T. Azuma, G. B.
Center for Emerging and Reemerging Infectious Diseases, Ministry of Nair, and D. E. Berg. 2000. Distinctiveness of genotypes of Helicobacter
Education, Culture, Sports, Science and Technology of Japan. P.K. is pylori in Kolkata, India. J. Bacteriol. 182:3219–3227.
a recipient of Senior Research Fellowship from University Grants 17. Myllyluoma, E., L. Veijola, T. Ahlroos, S. Tynkkynen, E. Kankuri, H.
Commission. Vapaatalo, H. Rautelin, and R. Korpela. 2005. Probiotic supplementation
improves tolerance to Helicobacter pylori eradication therapy—a placebo-
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