PESTICIDES-INDUCED OXIDATIVE
DAMAGE:
POSSIBLE PROTECTION
BY
Ahmed k. Salama and Omran A. Omran
Medical Laboratories Dept., Faculty of Science, Majmaah University,
Kingdom of Saudi Arabia, 2013
This work supported by the Essential and Health Sciences
Research Center , Scientific Research Deanship, Majmaah
University 1433-1434
THE PROBLEM:
Agrochemicals such as pesticides are used for achieving
better quality and quantity products but they have
many adverse effects on human. The toxic action of
pesticides may include the induction of oxidative stress
and accumulation of free radicals in the cell via
increasing the production of reactive oxygen species
(ROS), including hydrogen peroxide, superoxide, and
hydroxyl radicals. A major form of oxidative damage is
lipid peroxidation, which is initiated by hydroxyl free
radical through the extraction of hydrogen atom from
unsaturated fatty acids of membrane phospholipids
causing disturbance of the biochemical and
physiological functions of cells.
OBJECTIVES:
The aim of the study was planned to investigate:
1. The capability of the pesticides atrazine,
dimethoate and endosulfan to induce
oxidative stress in male rat following in vitro
treatment at different levels of each pesticide.
2. The possible protection of the oxidative
damage induced by these pesticides in rat
erythrocytes and hepatocytes using selenium
and a combination of vitamin E or vitamin C.
EXPERIMENTAL METHODS
Blood was obtained from rat by heart puncture and
then centrifuged at 3000 rpm for 5 min at 4°C. RBC’s
were taken and washed with phosphate buffered
saline, pH 7.2.
Liver was also dissected out and homogenized in
saline solution (1:10 w/v). The homogenates were
taken for treatment.
RBC's or liver homogenate treated with pesticides,
vitamins, selenium alone or with different
combinations and then incubated for 3 hours at
37°C in a shaking water bath. At the end of
incubation, all samples were subjected to
biochemical analysis.
BIOCHEMICAL ANALYSIS:
Lipid peroxidation level was determined and
expressed as nanomoles of malondialdehyde
(MDA)/mg protein
GSH content was determined and expressed as
µmole/mg protein.
Glutathione-S-transferase (GST-Px) activity was
estimated and expressed as units/mg protein.
Table (1): LPO, GSH and GSH-Px Levels of control (5% DMSO)
or treated erythrocytes and hepatocytes with VE, VC or Se.
LPO level
Treatment
(nmol MDA/mg protein)
RBC’s
Hepatocytes
GSH content
(µmole/mg protein)
GSH-Px Level
(µmol NADPH/min/mg
protein)
RBC’s
Hepatocytes
RBC’s
Hepatocytes
5% DMSO
8.96±0.19
12.54±0.96
52.55±6.10
233.14±0.88
0.69±0.10
2.14±0.88
VE (5mg)
8.55±0.11
11.84±0.11
60.22±9.10
244.70±10.15
0.85±0.11
2.80±0.11
VC (5mg)
9.16±0.23
13.22±0.77
55.17±8.88
255.00±10.70
0.77±0.23
2.22±0.77
8.92±0.44
13.16±0.90
60.10±7.56
247.30±10.20
0.90±0.40
2.30±0.10
8.90
12.69
57.01
245.04
0.80
2.37
Se (1mg)
Average
Table (2): LPO Level (nmoles MDA/mg protein) of treated
erythrocytes and hepatocytes with atrazine alone or combined
with VE, VC and/or Se.
LPO level in
Erythrocytes
Treatment
10mM
AT
% of Control
LPO level in
Hepatocytes
10mM
% of Control
12.10±0.76
135.96%
16.11±0.20
126.95%
AT + VE
9.57±0.16
107.53%
13.57±0.16
106.93%
AT + VE + Se
8.58±0.99
96.40%
11.20±0.46
88.26%
8.99±0.02
101.01%
12.52±0.22
98.66%
7.11±0.88
79.89%
11.44±0.60
90.15%
AT + VC
AT + VC + Se
Table (3): LPO Level (nmoles MDA/mg protein) of treated
erythrocytes and hepatocytes with dimethoate alone or
combined with VE, VC and/or Se.
LPO Level in
Erythrocytes
LPO Level in
Hepatocytes
Treatment
DM
DM + VE
DM + VE + Se
DM + VC
DM + VC + Se
10mM
% of Control
10mM
% of Control
10.20±0.10
114.61%
18.66±2.24
147.05%
9.10±0.40
102.25%
13.44±0.70
105.91%
8.11±0.80
91.12%
10.11±0.60
79.67%
8.13±0.40
91.21%
10.01±0.40
78.88%
7.11±0.40
79.89%
9.11±1.40
71.79%
Table (4): LPO Level (nmoles MDA/mg protein) of treated
erythrocytes and hepatocytes with endosulfan alone or
combined with VE, VC and/or Se.
LPO Level in
Erythrocytes
LP Level in
Hepatocytes
Treatment
ES
ES + VE
ES + VE + Se
ES + VC
ES + VC + Se
10mM
% of Control
10mM
% of Control
14.10±0.60
158.43%
18.10±0.76
142.63%
10.10±0.11
113.48%
15.10±0.44
118.99%
7.10±0.46
79.78%
11.10±0.22
87.47%
11.10±0.00
124.72%
13.10±0.88
103.23%
9.10±0.10
102.25%
11.10±0.16
87.47%
Table (5): GSH content (µmole/mg protein) of treated
.erythrocytes and hepatocytes with atrazine alone or combined
with VE, VC and/or Se.
Glutathione content in
Erythrocytes
Glutathione content in
Hepatocytes
Treatment
10mM
AT
AT + VE
AT + VE + Se
AT + VC
AT + VC + Se
% of Control
10mM
% of Control
100.80±12.80
176.81%
554.41±15.60
226.25%
88.10±5.10
154.53%
452.17±55.16
184.52%
40.18±6.19
70.47%
300.80±20.14
122.76%
60.19±5.02
105.58%
306.92±33.12
125.25%
33.00±4.68
57.88%
167.64±18.90
68.41%
Table (6): GSH content (µmole/mg protein) of treated
erythrocytes
.
and hepatocytes with dimethoate alone or
combined with VE, VC and/or S.
Glutathione content in
Erythrocytes
Glutathione content in
Hepatocytes
Treatment
10mM
DM
DM + VE
DM + VE + Se
DM + VC
DM + VC + Se
% of Control
10mM
% of Control
178.70±21.20
313.45%
516.77±18.24
210.89%
140.80±12.10
246.97%
377.14±16.90
153.91%
111.99±9.60
196.91%
299.10±18.50
122.06%
123.91±0.10
217.35%
254.01±20.10
103.66%
55.31±7.19
97.01%
77.91±0.10
31.79%
Table (7): GSH content (µmole/mg protein) of treated
.erythrocytes and hepatocytes with endosulfan alone or
combined with VE, VC and/or Se.
Treatment
Glutathione content in
Erythrocytes
10mM
ES
ES + VE
ES + VE + Se
ES + VC
ES + VC + Se
% of Control
Glutathione content in
Hepatocytes
10mM
% of Control
154.50±11.80
271.01%
456.10±44.16
186.13%
21.60±9.25
37.89%
331.10±18.14
135.13%
10.40±0.46
18.24%
144.20±14.29
58.85%
31.19±3.30
54.71%
279.19±12.89
113.94%
10.10±1.10
17.71%
104.10±9.10
42.48%
Table (8): Glutathione Peroxidase Level (µmoles NADPH/min/
.
mg protein) of treated erythrocytes and hepatocytes with
atrazine alone or combined with VE, VC and/or Se.
Treatment
GSH-Px level in
Erythrocytes
10mM
AT
AT + VE
AT + VE + Se
AT + VC
AT + VC + Se
% of Control
GSH-Px level in
Hepatocytes
10mM
% of Control
1.80±0.16
225.00%
5.41±0.20
228.27%
1.20±0.10
150.00%
3.17±0.16
133.76%
0.58±0.19
72.50%
1.80±0.66
75.95%
0.99±0.02
123.75%
0.92±0.12
38.82%
0.66±0.18
82.50%
0.64±0.60
27.00%
Table (9): Glutathione Peroxidase Level (µmoles NADPH/min/
.
mg protein) of treated erythrocytes and hepatocytes with
dimethoate alone or combined with VE, VC and/or Se.
Treatment
GSH-Px level in
Erythrocytes
10mM
DM
DM + VE
DM + VE + Se
DM + VC
DM + VC + Se
% of Control
GSH-Px level in
Hepatocytes
10mM
% of Control
1.80±0.20
225.00%
2.73±0.29
115.19%
1.10±0.10
137.50%
1.69±0.08
71.31%
0.31±0.11
38.75%
1.60±0.10
71.30%
0.81±0.10
101.25%
1.10±0.10
46.41%
0.11±0.10
13.75%
0.70±0.16
29.54%
Table (10): Glutathione Peroxidase Level (µmoles NADPH/min/
.mg protein) of treated erythrocytes and hepatocytes with
endosulfan alone or combined with VE, VC and/or Se.
Treatment
GSH-Px level in
Erythrocytes
10mM
ES
ES + VE
ES + VE + Se
ES + VC
ES + VC + Se
% of Control
GSH-Px level in
Hepatocytes
10mM
% of Control
2.10±0.40
262.50%
6.10±0.16
257.38%
1.10±0.21
137.50%
3.10±0.14
130.81%
0.40±0.46
50.00%
1.20±0.22
50.63%
1.19±0.01
148.75%
3.10±0.88
130.80%
0.90±0.10
112.50%
1.10±0.10
46.41%
CONCLUSION:
The results indicated that all treatments with
pesticides, enhanced the LPO level, GSH content and
GSH-Px activity via increasing oxidative stress in
erythrocytes and hepatocytes of male rats.
The treatment with selenium and a combination of
VE or VC was potentially reduced the free radicals
and declined the lipid peroxidation level, GSH
content and GSH-Px activity in erythrocytes or
hepatocytes and ameliorated the oxidative stress
induced by such pesticides and thus reduced the
lipo-peroxidative effect.
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