Transcript WP5.1 - Vollmar

The Interaction of Garlic With
Mediators of Inflammation
Prof. A. M. Vollmar
Dr. V. M. Dirsch
H - P Keiß
Overview
1.
2.
3.
4.
5.
Interaction of garlic powders and
constituents with activation of NF-B
Interaction of garlic powders and
constituents with expression of adhesion
molecules ICAM-1 & E-selectin
Interaction of garlic powders and
constituents with cytokine production
Additional targets
Outlook
Interaction of garlic powders and
constituents with NF-B

No inhibitory effect on TNF- induced NF-B activation was found
for:
–
–
–
–
–
–
–
–
–
–
Diallyl disulfide
Allyl mercaptane
S-allylcysteine
-glutamylcysteine
Allyl-methyl sulfoxide
Allicin
French garlic powders 2000
French garlic powders 2001
Spanish garlic powders 2000
Spanish garlic powders 2001
Interaction of garlic powders and
constituents with ICAM-1 & E-selectin

No inhibitory effect on TNF- induced expression of
adhesion molecules was found for:
– Diallyl disulfide
– Allyl mercaptane
– S-allylcysteine
Summary

Tested garlic compounds do not interact with NFB and expression of adhesion molecules
Garlic metabolites fail to inhibit the activation of the transcription
factor NF-kappaB and subsequent expression of the adhesion
molecule E-selectin in human endothelial cells
Verena M. Dirsch*, Hans-Peter Keiss, and Angelika M. Vollmar
Reviewer 1: I would like to suggest that the authors submit a new manuscript
providing additional experimental data on the effects of DADS and
AM on NF-B activity and E-selectin expression choosing longer
incubation periods. Furthermore, cytotoxicity data for DADS and
AM seem to be crucial.
Editor: ....“but I also would like to encourage you to submit a new mansucript when new
experimental findings have been added“
Eur. J. Nutrition
Interaction of garlic powders and
constituents with cytokine production in
whole blood
Allicin, DADS, AMSO2, -glutamyl-cysteine,
lipophilic and hydrophilic garlic extracts were
tested.
 Conflicting data between individual experiments

– Summer 2001: Il-1, TNF reduced, Il-10 increased
– Winter 2001/2002: Il-1, TNF and Il-10 increased
– Summer 2002: Il-1 reduced, TNF and Il-10 increased
Possible explanation

All sets represent self-contained observations and
are conclusive.
 Garlic powders used
– Summer 2001: French powders of year 2000
– Winter 2001/2002: French powders of year 2001 (cave
contaminated)
– Summer 2002: Mixed garlic powders from France and
Spain 2000 & 2001

For our publication we chose the data from
summer 2001, because data are conclusive (and
we have performed the luciferase assay)
Summary
Tested garlic compounds do interact with LPS-induced
cytokine liberation in human whole blood
Garlic (Allium sativum) modulates cytokine expression in LPSactivated human blood leading to an overall inhibitory
effect on NF-κB activity.
Hans-Peter Keiss, Verena M. Dirsch, Thomas Hartung, Thomas
Haffner ?? and Angelika M. Vollmar
Submitted to Journal of Nutrition
Additional targets
Garlic induced NO-liberation
NO - re le as e
x -fo ld
1 .5
S -allylc ys te ine
D iallyl d is ulfid e
A llylm e thyl-S O 2
A llic in
A llylm e thyl-s ulfid e
A llyl-m e rc ap tan
1 .0
0 .5
0 .0
CN
10 µM
100 µM
G arlic c o ns titue nts
NO - re le as e
1 .2 5
x-fold
1 .0 0
S pain 2000
0 .7 5
F ranc e 2000
S pain 2001
0 .5 0
F ranc e 2001
0 .2 5
0 .0 0
CN
50 µg/ml
P rintanor
500 µg/ml

DADS increased NO
liberation in HUVEC
“1.2” fold.
 All other tested garlic
constituents and
metabolites had no effect
on NO-liberation
 Some Garlic extracts even
reduced NO-liberation.
Garlic and PAI-1 and tPA
P rintano r S p ain 2 0 0 1
3000
2000

1000
All garlic extracts
– France 2000
0
CN
50
500
TNF
50
µg/m l
500
µg/m l
wo . T N F-
5 ng/m l T N F-
– France 2001
– Spain 2000
– Spain 2001
P rintano r s p ain 2 0 0 1
*
15
tPA [ng/m l]
P A I-1 [ng/m l]
***
showed no effect on
PAI-1 and tPA
liberation in HUVEC
10
5
0
CN
50
500
µg/m l
wo . T N F-
TNF
50
500
µg/m l
5 ng/m l T N F-
Garlic induced cytotoxicity
125
100
*
***
***
**
***
A MS O 2
75
A MS O
DADS
A c t.
***
50
***

25
.
ct
A
M
10
10
0µ
µM
M
1µ
M
10
0µ
µM
M
10
1µ
M
10
0µ
µM
10
1µ
C
M
N
0

125
100
***
***
***
***
75
F 2000
S 2000
50
S 2001
***
25
ct
A
/m
µg
.
l
l
0
50
µg
/m
µg
50
50
µg
0
50
/m
l
l
/m
l
/m
µg
50
0
µg
C
/m
N
l
0
50
P ercent
P ercent
**
***
Only DADS showed a
noteworthy effect on cell
viability
AMSO, AMSO2 and
garlic extracts had no
cytotoxic effects.
Outlook
Looking at vascular smooth muscle cell
proliferation and hypertrophy
as a target
for antiatherogenic compounds
The blood vessel
Endothelium
Smooth muscle cells
Connective tissue
Physiological function:
Intima
Media
Adventitia
vasoconstriction
vasorelaxation
Robbins pathologic basis of disease 1999
Vascular smooth muscle cells
and their response to injury
Endothelium
Smooth
Migration of smooth muscle cell
muscle cells to the
mitosis
intima
Elaboration of
extracellular
matrix
Intima
Media
Robbins pathologic basis of disease 1999
C: core of lipid; F: fibrous cap; L: llumen, moderately narrowed
Robbins pathologic basis of disease 1999
Primary rat aortic
vascular smooth
muscle cells (VSMC)
+ Serum
?
VSMC proliferation
DADS
Cell cycle, cell signaling
Angiotensin II - long-term effects
Ang II
AT1-Receptor


Cell growth
Cell migration
ECM deposition
Growth factor and cytokine production
Intimal thickening, vascular remodeling,
inflammation
Primary rat aortic
vascular smooth
muscle cells (VSMC)
+ Ang II
Hypertrophy
?
DADS
Cell signaling
Relevant publications
Haider UGB, Sorescu D, Griendling KK, Vollmar AM, Dirsch VM.
Resveratrol suppresses angiotensin II-induced Akt/Protein kinase B as well as
p70 S8 kinase phosphorylation and subsequent hypertrophy in rat aortic
smooth muscle cells. Mol Pharmacol 62: 772-777, 2002
Haider UGB, Sorescu D, Griendling KK, Vollmar AM, Dirsch VM.
Resveratrol increases serine15-phosphorylated but transcriptionally impaired
p53 and induces a reversible DNA replication block in serum-activated vascular
smooth muscle cells. Mol Pharmacol, in press