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Institute for Applied Biosciences, Ben-Gurion University of the Negev, Beer Sheva, Israel
Key words: propolis extract, herpes simplex virus, antiviral activity, plaque assay, cytopathic effect
Abstract
Propolis, a natural product from beehives, comprises a complex of chemicals, the most important group being
flavinoids, which play a role in antiviral protection.
To test the inhibitory effect of propolis extract against
herpes simplex viruses in vitro and in vivo.
propolis was added to Vero cells at various
times and concentrations before, at or after infection with HSV-1. In
vivo the effect of propolis was tested in newborn rats infected s.c. or
i.p. and on rabbit corneas infected with HSV-1.
In vitro 0.5% propolis extract caused 50% inhibition of
HSV infection. There was indirect evidence for a strong interaction
between the propolis extract and the surface of the Vero cells, but there
was no direct interaction with HSV-1 particles. Administration of
propolis before or at the time of infection yielded the most significant
inhibitory effect, but even when 10% propolis extract was added 2
hours post-infection it gave 8085% protection. In vivo as little as 5%
propolis prevented the appearance and development of symptoms of
local and i.p. HSV-1 infection in rats and of corneal HSV-1 infection in
rabbits. There were no cytotoxic effects at a concentration of 10% in
vitro or 20% in vivo.
The potent antiviral activity of propolis against HSV1 infection in vitro and In vivo is probably due to prevention of virus
absorption into the host cells and/or inhibition of an internal step(s)
during the viral replication cycle.
Background:
Objectives:
Methods: In vitro:
Results:
Conclusions:
IMAJ 2002;4(Suppl):923927
African green monkey kidney (Vero) cells were purchased from the
American Type Culture Collection, Rockville, MD, USA. Cells were
grown in RPMI medium containing 10% fetal calf serum, 1%
glutamine, antibiotics and incubated at 378C in humidified air
containing 5% CO2. HSV-1 and HSV-2 were also obtained from the
American Type Culture Collection. HSV-1 stock (3.106 pfu/ml) was
used for infecting the cells and the animals. HSV-2 stock (1.106 pfu/
ml) was used only for infecting cells.
Animals
The experiments were carried out on white, newborn (23 day old)
rats and young male (1.52 kg) rabbits. All animal experiments were
conducted in accordance with the ethical guidelines of the
Committee for the Ethical Care and Use of Laboratory Animals of
Ben-Gurion University of the Negev.
Propolis
In Vitro experiments
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Animal experiments
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Table 1.
Group of
animals
Statistical analysis
Non-infected,
non-PE treated
Non-infected,
PE treated
Infected,
non-PE treated
Infected,
PE treated
Appearance
of symptoms
Treatment Days %
route
(p.i.)
Death
Days
(p.i.)
Infective
virus
%
NT
s.c.
i.p.
0
0
98+2
7-9
0
0
98+2
NT
NT
98+2
s.c.
i.p.
56
5
10+0.5
5+0.3
14
10
10+0.5
5+0.3
10+0.5
5+0.3
The number of animals presenting symptoms and subsequently dying was expressed as the percentage
of the total number of tested animals. A total of 40 animals were used for each treatment. Infective virus
(collected in swabs from infected areas 3 days p.i.) was tested in cell culture. The number of animals
containing infective virus was expressed as a percentage of the total number of animals tested.
p.i.= post infection, NT = Not tested. Data are mean + SD (n = 40).
Original Articles
Table 2.
Group of
animals
Appearance
of symptoms
Treatment Days
%
route
(p.i.)
Death
Days
(p.i.)
Infective
virus
%
Non-infected,
0
NT
non-PE treated
Non-infected,
Drops
0
NT
PE or Zovirax i.v.
0
NT
treated
Infected,
45
100
1012
100
100
non-PE treated
Infected,
Drops
9*
35+2
0
35+2
PE treated
i.v.
10**
30+2
0
30+2
Infected, treated Ointment 78
40+3
0
40+3
with Zovirax
The number of animals containing infected virus was expressed as the percentage of the total
number of animals tested. A total of 10 rabbits were used for each treatment. Infective virus
(collected in swabs from infected eyes 3 days p.i.) was tested in cell culture.
* Light keratitis with secretions
** Reversible light keratitis
Data are mean + SD (n = 10).
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What's important is what we become, not what we have. It's how well we've
learned to serve. It's the relationships we've built.
Anonymous
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