Phytomedicine 14 (2007) 285–288
An extract of Pelargonium sidoides (EPs 7630) inhibits in situ adhesion of
Helicobacter pylori to human stomach
N. Wittschiera, G. Fallerb, A. Hensela,?
aInstitute for Pharmaceutical Biology and Phytochemistry, University of Mu ¨nster, Hittorfstrasse 56, D-48149 Mu ¨nster, Germany
bSt. Vincentius-Hospital, Institute for Pathology, Su ¨dendstraße 37, D-76 137 Karlsruhe, Germany
Received 21 July 2006; accepted 3 November 2006
Root extract from Pelargonium sidoides DC is used therapeutically as antimicrobial agent against infections of the
respiratory system. In order to elucidate possible modes of actions we investigated the influence of P. sidoides root
extract on microbial adhesion with Helicobacter pylori as model microorganism, a germ with a strong adherence to
human stomach tissue. In an in-situ anti-adhesion assay intact human stomach tissue from patient resectates was
incubated with fluorescent-labelled bacteria. Epithelial adhesion occurred in untreated samples and was quantified by
fluorescent microscopy. Pre-treatment of the bacteria with Pelargonium extract showed good anti-adhesive activity.
The antiadhesive effect was clearly dose-dependent in a range from 0.001 to 10mg/ml. Within agar diffusion-test the
extract had no direct cytotoxicity against H. pylori. The results show that the root extract from Pelargonium sidoides is
a potent anti-adhesive agent against H. pylori and could therefore be a useful choice to avoid the first step of a bacterial
r 2006 Elsevier GmbH. All rights reserved.
Keywords: Pelargonium sidoides; EPs 7630; Helicobacter pylori; Adhesion; Human stomach
Root extracts from Pelargonium sidoides DC (Ger-
aniaceae) accomplish more and more for the treatment
of acute respiratory infections. The phytochemical
profiling has indicated the dominating presence of
mainly polymeric proanthocyanidins and monomeric
flavan-3-ols beside considerably amounts of coumarins
with a high degree of oxygenation in the aromatic ring
system. Phenolic acids with gallic acid as lead structure
are found beside minor amounts of quercetin and
sitosterol-glucoside (Kolodziej, 2000; Kolodziej and
Kayser, 1998). A proprietary root extract, referred to
as EPs 7630, has been registered in several countries for
treatment of infections of respiratory system. Despite
sufficient clinical data the respiratory mode of action for
the extracts is not fully understood. From the present
knowledge a moderate antimicrobial activity (Kayser
and Kolodziej, 1997) and immunmodulatory effects
(Kayser et al., 2001) via increased NK-cell formation
and TNF-a-, iNO- and INF-b release (Koch et al., 2002)
are focused as the main therapeutic principles. Looking
at the antimicrobial effects it gets obvious that the
minimal inhibitory concentrations for the potentially
active compounds are in a range higher than 0.2mg/ml
and also the extracts exhibit MICs in a range from 0.6 to
ARTICLE IN PRESS
0944-7113/$-see front matter r 2006 Elsevier GmbH. All rights reserved.
?Corresponding author. Tel.: +492518333380.
E-mail address: email@example.com (A. Hensel).
investigations have revealed slight antimicrobial activ-
ities against Mycobacterium, mediated by the unsatu-
rated fatty acids with MIC of 2mg/ml (Seidel and
Taylor, 2004). From the clinical point of view it is not
very reasonable that under therapeutic conditions
antimicrobial activity at such high MIC values will be
very pronounced due to direct bacteriostatic or bacter-
iocidic effects. For the good clinical effects reported
more indirect mechanisms may be responsible. A hint
into this direction may be the report of Dorfmu ¨ ller et al.
(2005), showing that EPs 7630 inhibits the internaliza-
tion of Streptococcus pyogenes into Hep-2 cells under in
vitro conditions. From our point of view the adhesion of
many pathogens to host epithelial cells or host tissue is a
prerequisite for starting a virulent process. It has to be
proven if Pelargonium extract has potential antiadhesive
capacities, which might explain many of the observed
antibacterial effects. A good model for studying
adhesive and antiadhesive processes is Helicobacter
pylori for which different receptor-like adhesins on the
outer cell wall are described in detail (Odenbreit, 2005),
interacting with complementary structures on the hu-
man stomach epithelium. This prototype of a strongly
adhesive microorganism can serve to obtain detailed
information on potential antiadhesive effects of test
compounds and many data obtained in this model can
be transferred to the adhesion strategy of other germs.
Aim of the following study was to test EPs 7630 in this
in situ system and to quantitate potential antiadhesive
Materials and methods
EPs 7630 was obtained by ISO Arzneimittel, Ettlin-
gen, Germany. Adhesion and antiadhesion tests were
performed according Lengsfeld et al. (2004a,b). In
principle, FITC-labelled bacteria are preincubated with
the test compounds. Deparaffinated sections from hu-
man stomach are incubated with the labelled bacteria.
counted under blinded conditions by fluorescent micro-
scopy and evaluated against a non-treated control.
Maximal adhesion, seen in the untreated control groups
(negative control) is expressed as total 100% adhesion
with a score (+++++). Lower adhesion is indicated
by (++++, +++, ++, +, ?) with missing (?)
adhesion being found in the positive control (sialyl
lactose). Results presented are the mean of three
independent experiments. Additionally fluorescent area
intensity was calculated by ImageJssoftware (Olympus,
Germany), standardizing fluorescent area of the nega-
tive control as 100%. To exclude unspecific cell toxicity
of test compounds against H. pylori a disk diffusion test
was performed at 2.5mg/ml of test compounds using
to epithelial tissueare
BD Sensi-Disks (Becton and Dickinson, Germany),
placed on agar plates.
Results and discussion
In a validated (Lengsfeld et al, 2004b) in situ assay,
based on the model of Falk et al. (1993), fluorescent-
labelled H. pylori was incubated together with histolo-
gical sections from human stomach tissue. Microorgan-
isms sticking to the epithelia via receptor-mediated
adhesion after extensive washing can be counted by
fluorescent-microscopy techniques. It got obvious, that
within a few minutes after starting the incubation
exclusively epithelial adhesion was visible (Fig. 1A).
No interaction of H. pylori to connective and muscle
tissue was observed, which is in good accordance with
data, that adhesin ligands for H. pylori are exclusively
located on the luminal side of stomach epithelium. This
adhesion was assessed within a semi-quantitative,
blinded evaluation as maximal (+++++) and the
fluorescent area was standardized on 100%. For positive
controls bacteria were preincubated with 3-sialyllactose,
leading to a nearly complete inhibition of adhesion.
Additionally plant-derived extract from Abelmoschus
esculentus was used as a further positive control with
inhibition rates 490% and counted with no fluores-
cence over the endogenous background. When using
EPs 7630 extract in different concentrations a clear
dose-dependent antiadhesive activity was obvious, with
concentrations of 1mg/ml being as active as the positive
controls (Table 1). To exclude any direct cytotoxic
effects against H. pylori a disk diffusion test was
performed with all EPs 7630 extract concentrations as
used in the adhesion tests. While the positive control
amoxicillin (0.5mg) had significant zones of inhibition
(2500mm) no signs of growth inhibition was observed
for EPs 7630, even at 10mg/ml, indicating absence of
any direct cell toxicity at the used concentration.
From these results it gets obvious that Pelargonium
sidoides extract has significant antiadhesive effects
against H. pylori. The bacterial adhesins, located on
the outer cell wall are responsible for interaction with
mucosal glycoproteins and epithelial mucins and are
blocked or inactivated by extract compounds. Because it
was shown in previous studies that these receptor-
systems can easily antagonized by exogenous polymeric
compounds (e.g. polysaccharides) by mimicking endo-
genous ligands it may be worth to investigate EPs 7630
on the occurrence of carbohydrates. Another possibility
concerning active compounds may be the polymeric
proanthocyanidins, leading to an unspecific interaction
by H-H-linkages with the bacterial adhesins, blocking
them by a kind of tannin-like adstringent process.
Summarizing we assess P. sidoides root extract to be a
ARTICLE IN PRESS
N. Wittschier et al. / Phytomedicine 14 (2007) 285–288286
strong antiadhesive with the potential to be investigated
deeper concerning this mode of action.
Dorfmu ¨ ller, A., Jung, I., Tioua, D.L., Engels, I., Daschner,
Wirkmechanismen des Pelargonium-sidoides-Wurzelextraktes
EPs7630: Hemmung der Interaktion von A-Streptokokken
und HEp-2-Zellen. Zt. Phytother. 26 (Suppl.), S6–S7.
Falk, P., Roth, K.A., Bore ´ n, T., Westblom, T.U., Gordon,
J.I., Normark, S., 1993. An in vitro adherence assay reveals
that Helicobacter pylori exhibits cell lineage specific tropism
in the human gastric epithelium. Proc. Natl. Acad. Sci.
USA 90, 2035–2039.
Kayser, O., Kolodziej, H., 1997. Antibacterial activity of
extracts and constituents of Pelargonium sidoides and
Pelargonium reniforme. Planta Med. 63, 508–510.
Kayser, O., Kolodziej, H., Kiderlen, A.F., 2001. Immunmo-
dulatory principles of Pelargonium sidoids. Phytother. Res.
Koch, E., Lanzendo ¨ rfer-Goossen, S.H., Wohn, C., 2002.
Stimulation of interferon (INF)-b-synthesis and natural
liller (NK) cell activity by an aqueous-ethanolic extract
from roots of Pelargonium sidoides (Umckaloabo). Nau-
nyn-Schmiedeberg’s Arch. Pharmacol. 365 (Suppl. 1), R75
ARTICLE IN PRESS
Fig. 1. Fluorescent microscopy (200?) of representative in situ experiment with FITC-labelled H. pylori on human gastric mucosa:
(A) complete adhesion (+++++) of non-treated bacteria (negative control), (B) positive control (?), (C) Pelargonium sidoides
root extract EPs 7630 1mg/ml, and (D) Pelargonium sidoides root extract EPs 7630 0.1mg/ml.
of FITC-labelled H. pylori to sections of human gastric mucosa
Effect of a 2h pre-treatment with different concentrations of Pelargonium sidoides root extract EPs 7630 on the adhesion
Pre-treatment of bacteria Bacterial adhesion after blinded
fluorescent microscopic evaluation
Fluorescent area (%) related to
the negative control
Untreated (negative control)
30-/60-Sialyllactose (positive control)
Abelmoschus esculentus extract (positive control)
EPs 7630, 10mg/ml
EPs 7630, 1mg/ml
EPs 7630, 0.5mg/ml
EPs 7630, 0.1mg/ml
EPs 7630, 0.01mg/ml
Quantification of bacterial adhesion: ? almost no binding; + very weak binding; +++++ very strong, maximal binding.
N. Wittschier et al. / Phytomedicine 14 (2007) 285–288 287
Kolodziej, H., 2000. Traditionally used Pelargonium species: Download full-text
chemistry and biological
extracts and their constituents. Curr. Topics Phytochem. 3,
Kolodziej, H., Kayser, O., 1998. Pelargonium sidoides DC. Zt.
Phytother. 19, 141–151.
Lengsfeld, C., Deters, A., Faller, G., Hensel, A., 2004a. High
molecular weight polysaccharides from black currant seeds
inhibit adhesion of Helicobacter pylori to human gastric
mucosa. Planta Med. 70, 620–626.
Lengsfeld, C., Titgemeyer, F., Faller, G., Hensel, A., 2004b.
Glycosylated compounds from okra inhibit adhesion of
Helicobacter pylori to human gastric mucosa. J. Agric.
Food Chem. 52, 1495–1503.
Odenbreit, S., 2005. Adherence properties of Helicobacter
pylori: impact on pathogenesis and adaption to the host.
Int. J. Med. Microbiol. 295, 317–324.
Seidel, V., Taylor, P.W., 2004. In vitro activity of extracts and
constituents of Pelargonium against rapidly growing
mycobacteria. Int. J. Antimicrob. Agents 23, 613–619.
ARTICLE IN PRESS
N. Wittschier et al. / Phytomedicine 14 (2007) 285–288 288