activity of proton pump inhibitors.

1. Invitro Activity of
PPIs on mouse
macrophage
RAW264.7 cell
line
Lansoprazole
,Omeprazol,Esmoprazol
In Vivo Activity
of Omeprazol
on
carrageenan-
induced rat paw
edema
LPS and H.pylori induced TNF-αand IL-
1βproductionco-incubated with
human neutrophils and human
umbilical cord endothelial cells
inhibit the H. pylori extract induced
expression of CD11b and CD18 on
neutrophils and neutrophil-
dependent adhesion to endothelial
Anti-oxidant at and anti-
inflammatory efficacy at
an intraperitoneal dose of
2.5 mg/kg
INTRODUCTION
PPIs omeprazole, lansoprazole,
pantoprazole, esomeprazole,
rabeprazole were purchased
Swapnroop chemicals, Aurangabad,
India.
Lambda carrageenan
(22049 Sigma) and Thiazolyl Blue
Tetrazolium Blue – MTT (M 2128
Sigma),
Diclofenac sodium was
generously supplied by Lord Venky
Pharma Private Limited, Yanam,
India.
ELISA kits for
determination of mouse TNF-α, IL-6
and IL-1β were purchased from
eBioscience (San Diego, USA).
Macrophage cell line,
RAW 264.7 (ATCC TIB-71), was
obtained from the American Type
Culture Collection, Manassas, VA,
USA.
MATERIAL EXPERIMENTAL PROTOCOL
Invitro Technique Invivo Technique
Experimental Animl group
Adult spargue
Dawley Male Rat ,11
treatment groups
containing 6 rats
each. The acute in
vivo antiphlogistic
activity of PPIs was
tested at 15, 30 and
60 mg/kg oral doses.
Orally administered
diclofenac sodium (5
mg/kg) served as a
standard drug.
Nitric
Oxide
Measurme
ment
Carrageenan
Induced paw
inflammation
and Mechanical
Hyper Algesia
RESULT
0
10
20
30
40
50
60
70
80
90
100
110
10 20 50 100 200 300 400
%Survival
Conc of drug in uM
Effect of different PPIs on % survival of RAW 264.7 cells
OPZ LPZ PPZ RBZ EMZ
Name of drug Omeprazole Lansoprazole Pantoprazole Rabeprazole Esomeprazole
IC 50 341.3 uM 149 uM 498 uM 146 uM 420 uM
% survival at 50
uM
100 80 82 85 99
% Survival at
400 uM
39 14 40 15 51
PPIs inhibit the LPS-induced cytokine release from RAW-264.7 cells
% survival at 50
uM
100 80 82 85 99
% Survival at
400 uM
39 14 40 15 51
PPIs inhibit carrageenan-induced acute paw inflammation in rats
1 h r 4 h r 1 h r 4 h r 1 h r 4 h r 1 h r 4 h r
0
5
10
15
20
25
30
35
40
45
O PZ LPZ PPZ EPZ RPZ
1 5 m g /k g 3 0 m g /k g 6 0 m g /k g
N orm al C arrag een an S tandard
@
a
a
b
b
b
b
b
b
c
bb
c
c c
b
***
***
***
**
***
** c
***
***
***
***
***
***
***
***
**
@
%Riseinpawvolume
Figure 1: PPIs inhibit carrageenan induced acute paw
inflammation in rats
15mg/kg 30mg/kg 60mg.kg
0
5
10
15
20
25
30
35
40
45
50
55
Normal Carrageenan Standard OPZ LPZ PPZ EPZ RPZ
@
a a a
a a
a a a a
b b
c
b b b
c
Treatment
Pawwithdrawalthreshold(g)
1 hr 4 hr 1 hr 4 hr 1 hr 4 hr 1 hr 4 hr
0
5
10
15
20
25
30
35
40
45
OPZ LPZ PPZ EPZ RPZ
15 mg/kg 30 mg/kg 60 mg/kg
Normal Carrageenan Standard
@
a
a
b
b
b
b
b
b
c
bb
c
c c
b
***
***
***
**
***
** c
***
***
***
***
***
***
***
***
**
@
%Riseinpawvolume
Effects of PPIs on the viability (% Survival) of RAW264.7 murine macrophages. Cells were incubated with various
concentrations of PPIsfor 24 hr, and cell viability was determined by MTT assay.The results are expressed as
means±SD. P<0.0001.
N
orm
al
C
arrageenanStandard
O
PZ
LPZ
PPZ
EPZ
R
PZ
0
20
40
60
80
100
120
Treatment
Tnf-level(pg/mgofprotein)
Effect of PPIs on TNF-α level in Paw tissue
supernatant of carrageenan injected animal.
N
orm
al
C
arrrageenanS
tandard
O
P
Z
LP
Z
P
P
Z
E
P
Z
R
P
Z
0
5000
10000
15000
20000
25000
a*
a*
a*b*
a*
a* a*b*
Treatment
IL-6level(pg/mgofprotein)
Effect of PPIs on IL-6 level in Paw
tissue supernatant of carrageenan
injected animal.
N
om
al
C
arrageenanS
tandard
O
P
Z
LP
Z
P
P
Z
E
P
Z
R
P
Z
0
5000
10000
15000
20000
25000
a*
a*
a*b*
a*
a* a*
Treatment
IL-1level(pg/mgofprotein)
Effect of PPIs on IL-1β level in Paw tissue
supernatant of carrageenan injected
animal
DISCUSSION AND CONCLUSION
Both In-vivo and In-vitro anti-inflammatory activity of Proton Pump Inhibitor was investigated. There were three major findings
in our study. First, PPIs reduced the rise in paw volume, by dose dependent manner and also reduced the pain in inflamed
condition. Second, the suppression of NO production at the initial stage of inflammation and third, in RAW 264.7 cell line PPIs
exert anti-inflammatory effects through inhibition of the production of pro-inflammatory cytokine TNF-α, IL-1β and IL-6 from LPS
stimulated macrophages
In conclusion, the overall results of this study clearly indicate that the Proton pump inhibitor had higher anti-inflammatory and
cytokine inhibitory action at increasing dose. Although the exact mechanism(s) of Proton pump inhibitors on suppressing the
cytokine were not investigated in the present study, this could constitute an area of future studies
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In-vitro and In-vivo anti-inflammatory activity of proton
pump inhibitors
Department of Pharmacology,(contact no.-9158830005, email adress-sanketgandhi111@gmail.com)
R. C. Patel Institute of Pharmaceutical Education &
Research, Shirpur- 425405, Maharashtra, India
Sanket Gandhi, Jayesh Patil, Umesh B. Mahajan, Sameer N. Goyal, Chandragouda R. Patil*
Effect of PPIs on viability of
RAW 264.7 cell
By MTT
Assay
Effect of PPIs on Cytokinin
release from RAW 264.7 cell
cytokines
levels
TNF-a,
IL1 and
IL6 levels
by ELISA