Bull Vet Inst Pulawy 48, 105-108, 2004
EFFECTS OF VITAMIN E AND PREDNISOLONE
ON BIOCHEMICAL AND HAEMATOLOGICAL PARAMETERS
IN ENDOTOXAEMIC NEW ZEALAND WHITE RABBITS
ENVER YAZAR1, RAMAZAN COL2, SIBEL KONYALIOGLU3,
YAVUZ OSMAN BIRDANE4, MUAMMER ELMAS1 AND AHMET LEVENT BAS1
Department of Pharmacology and Toxicology1,
Department of Physiology2, Faculty of Veterinary Medicine,
University of Selcuk, 42031, Konya, Turkey
Department of Biochemistry3, Faculty of Pharmacia,
University of Ege, 35100, Izmir, Turkey
Department of Pharmacology and Toxicology4,
Faculty of Veterinary Medicine, University of AKU, 03200, Afyon, Turkey
e-mail: [email protected]
Received for publication December 16, 2003.
Abstract
Effects of prednisolone and vitamin E on
biochemical and haematological values were investigated in
endotoxaemic rabbits. Forty rabbits were used and divided
into four equal groups. Group 1 served as the control group;
group 2 was infused with lipopolysaccharide (LPS) for 6 h;
group 3 was injected with prednisolone before LPS
administration; group 4 was injected with vitamin E for 4
consecutive days before LPS administration. Serum and
blood samples were collected 8 h after the onset of LPS
injection. Serum myoglobin, alanine aminotransferase,
gamma glutamyl transferase, amylase, total bilirubin, direct
bilirubin, creatinine, blood urea nitrogen, albumin, globulin,
total protein, cholesterol, total lipids, triglycerides, low
density lipoprotein, very low density lipoprotein, sodium,
potassium and magnesium contents were measured. Red
blood cell, white blood cell, platelet counts and percentage of
differential leukocyte values were determined. It was found
that prednisolone and vitamin E had a protective effect in
endotoxaemic shock. Prednisolone was more effective in
endotoxaemia than vitamin E.
Key words: rabbits, lipopolysaccharide,
prednisolone, vitamin
haematological values.
E,
biochemical
cascades. The endotoxic shock causes high mortality in
intensive care patients (2, 4, 13).
Corticosteroids decrease plasma levels of
inflammatory cytokines. During entotoxaemia,
cytokines are released from phagocytic cells and may
cause endothelial damage, leukocyte adherence to
vessel walls and consumptive coagulopathy leading to
vasodilatation, vascular leakage and refractory
hypotension. Also, prednisolone decreases mortality
rate in endotoxic shock (16, 17, 19).
Vitamin E (alpha-tocopherol), as an
antioxidant, protects the cell from reactive oxygen
radical damage. Oxygen derived free radicals are
important mediators of cellular injury in acute phase
sepsis (2).
Changes in biochemical and haematological
parameters are observed in specific organ failure or
damage. In this study, the possible protective effects of
vitamin E and prednisolone on biochemical and
haematological values in endotoxaemia were
investigated.
values,
Severe Gram-negative bacterial infections lead
to development of the endotoxic shock, a condition
characterized by fever, tachycardia, tachypnea,
hypotension, disseminated intravascular coagulation,
multiple
organ
failure,
and
even
death.
Lipopolysaccharide (LPS), originating from the cell
wall of Gram-negative bacteria, causes the endotoxic
shock. LPS, released during bacterial lysis, stimulates
the production of arachidonic acid metabolites,
complement factors, cytokines, and coagulation
Material and Methods
Forty New Zealand white male rabbits (12-16
months of age, weighing 2-2.5 kg, from the Veterinary
Research Institute, Adana) were randomly divided into
four equal groups. Group 1 (control group) received
intravenously physiological saline solution; group 2
was infused with E. coli lypopolysaccharide (LPS E.
coli 0111;B4, Sigma, 10 ml/h, 100 µg/kg/h by the
marginal ear vein for 6 h) (12); group 3 was injected
subcutaneously with 10 mg/kg of prednisolone (PR,
Prednisolon® amp, Fako, Istanbul, Turkey) 30 min
before the infusion of LPS (20); group 4 was injected
intraperitoneally with 10 mg/kg of vitamin E (VE, DL-
106
alpha tocopherol acetate, Evigen® amp, Aksu Farma,
Istanbul, Turkey) for 4 successive days before LPS
administration (21). Blood samples were taken through
a catheter inserted into a femoral artery 8 h after
starting the LPS infusion. Blood and serum samples
were collected to determine haematological and
biochemical parameters.
Serum alanine aminotransferase (ALT),
gamma glutamyl transferase (GGT), amylase, total
bilirubin (Tbil), direct bilirubin (Dbil), creatinine,
blood urea nitrogen (BUN), albumin, globulin, total
protein (TP), cholesterol, total lipids (Tlipid),
triglycerides, low density lipoprotein (LDL) and very
low density lipoprotein (VLDL) levels were measured
with an auto-analyser (Olympus AU 560). Serum
myoglobin level was measured with Abbott Axsym
system, sodium (Na) and potassium (K) contents were
determined with flame photometer, and magnesium
(Mg) concentration was measured with Hitachi 911.
Red blood cell (RBC), white blood cell (WBC), and
platelet counts were obtained with a haemocytometer.
Percentage of differential leukocyte values were
determined in blood smears.
All the values are expressed as mean ± SE.
The results were analysed by Tukey multiple range test
(SPSS for Windows, release 6.0). In all cases,
probability of error of less than 0.05 was chosen as the
criterion of statistical significance.
Results
Effects of VE and PR on biochemical and
haematological parameters are given in Tables 1 and 2,
respectively. In biochemical parameters, LPS caused
statistically significant increases in myoglobin, GGT,
ALT, Tlipid, triglyceride, VLDL, Mg, and K levels
(Table 1). Increases in these parameters were
significantly suppressed by the administration of PR
except for GGT, and VE except for Tlipid, triglyceride
and VLDL. PR caused statistically significant increase
in RBC and WBC counts (Table 2). However, in all
experimental groups, platelet counts decreased in a
statistically significant way (Tables 1 and 2).
Discussion
Increases in levels of biochemical values are
generally observed due to organ failure or damage. In
the present study, LPS caused increases in myoglobin,
GGT, ALT, Tlipid, triglyceride, VLDL, Mg and K
levels and these increases were significantly
suppressed by the administration of PR except for
GGT. On the other hand, increases in levels of
myoglobin, GGT, ALT, Mg and K were significantly
suppressed by the administration of VE except for
Tlipid, triglyceride and VLDL. It may be stated that
VE has no protective effect on lipid metabolism in
endotoxaemia. However, PR induced more protective
effect on the parameters (except for GGT) altered in
endotoxaemia than VE. It has been reported that LPS
caused increases in ALT, GGT, triglyceride, VLDL,
and potassium levels (1, 5, 7, 9, 15, 19) and that the
increase in ALT level following administration of LPS
was suppressed by PR (20). Increases in serum ALT
and GGT activities are supposed to be of hepatic
damage (3, 19).
Table 1
Effects of vitamin E and prednisolone on biochemical parameters in endotoxaemic rabbits (mean±SE)
Myoglobin ng/ml
Creatinine mg/dl
BUN mg/dl
Albumin g/dl
Globulin g/dl
TP g/dl
GGT U/L
ALT U/L
Amylase U/L
Tbil mg/dl
Dbil mg/dl
Cholesterol mg/dl
Tlipid mg/dl
Triglyceride mg/dl
LDL mg/dl
VLDL mg/dl
Mg mg/dl
Na mmol/l
K mEq/L
Control
(n=10)
536±109 a
1.27±0.09 a
31.7±2.03 a
2.82±0.07 a
1.58±0.07 a
4.42±0.11 a
3.88±0.42 a
87.3±15.2 a
207±21.8 a
0.12±0.02 a
0.03±0.01 a
41.2±6.58 a
171±19.8 a
93.8±17.1 a
20.4±3.97 a
18.7±3.39 a
3.12±0.27 a
132±3.63 a
4.42±0.26 a
LPS
(n=10)
1424±311 b
1.84±0.13 a
28.8±2.07 a
2.80±0.19 a
1.57±0.15 a
4.33±0.29 a
21.2±5.83 b
213±22.6 b
249±15.3 a
0.09±0.01 a
0.03±0.01 a
32.7±3.37 a
393±46.2 b
302±37.4 b
36.6±7.89 a
65.5±8.12 b
5.58±0.34 b
137±2.81 a
9.29±1.98 b
LPS+PR
(n=10)
824±109 ab
1.75±0.28 a
40.8±4.5 a
2.96±0.09 a
1.70±0.16 a
4.46±0.16 a
17.8±13.1 b
153±19.3 ab
242±38.4 a
0.13±0.03 a
0.03±0.01 a
37.2±6.83 a
275±46.2 ab
205±39.7 ab
22.1±5.59 a
40.6±7.93 ab
4.23±0.47 ab
137±3.59 a
7.64±1.22 ab
LPS+VE
(n=10)
585±79 a
1.48±0.24 a
43.2±5.86 a
2.85±0.13 a
1.82±0.15 a
4.67±0.25 a
13.0±2.49 ab
122±21.8 a
225±77.9 a
0.12±0.05 a
0.02±0.01 a
50.8±10.9 a
383±67.3 b
300±56.2 b
30.7±5.60 a
73.5±16.4 b
3.05±0.41 a
138±4.34 a
4.59±0.32 a
a,b; Differences in the same row are statistically significant when the values are marked with different letters (P<0.05).
LPS, lipopolysaccharide, PR; prednisolone, VE; vitamin E.
107
Table 2
Effects of vitamin E and prednisolone on haematological parameters in endotoxaemic rabbits (mean±SE)
RBC 106 mm3
WBC 103 mm3
Platelet 103 mm3
Basophils %
Eosinophils %
Neutrophils %
Lymphocytes %
Monocytes %
Control
(n=10)
4.46±0.24 ab
7820±1306 ab
381±18.7 a
1.10±0.23 a
1.70±0.21 a
63.1±3.19 ab
30.5±3.06 a
3.60±0.40 a
LPS
(n=10)
3.77±0.22 a
4530±4320 a
94.9±6.87 b
0.60±0.16 ab
1.10±0.23 ab
33.9±2.72 c
61.2±2.59 b
3.20±0.41 ab
LPS+PR
(n=10)
5.23±0.31 b
9430±1300 b
242±13.9 c
0.30±0.15 b
0.40±0.16 b
71.1±2.75 a
27.4±2.88 a
0.80±0.20 c
LPS+VE
(n=10)
4.15±0.20 a
2390±3585 a
180±8.81 d
0.50±0.16 ab
0.70±0.15 b
57.6±3.84 b
39.1±3.77 a
2.10±0.58 bc
a,b,c; Differences in the same row are statistically significant when the values are marked with different letters
(P<0.05). LPS, lipopolysaccharide, PR; prednisolone, VE; vitamin E.
In the present study, LPS increased serum
levels of total lipid, triglyceride and VLDL. In
endotoxaemia, tumour necrosis factor level is increased,
and this increase may suppress lipolytic enzymes, which
convert serum VLDL and triglyceride rich proteins to
LDL and HDL. This suppression may raise serum levels
of triglyceride and VLDL (14). In the present study,
hyperkalaemia and hypermagnesaemia were observed
after LPS administration, and the results were in
agreement with other studies (10, 15). These increases
were suppressed by administration of PR and VE.
LPS
causes
disseminated
intravascular
coagulation, which is associated with coagulation
disorders and loss of platelets. In this study,
administration of LPS decreased platelet counts, and this
statistically significant decrease was observed in PR and
VE groups as well, which is in agreement with previous
studies (1, 6, 11, 18, 20, 21). In addition to this, LPS
decreased percentage of neutrophils and increased
percentage of lymphocytes compared to other groups. It
was found out that PR administration in endotoxaemia
caused a statistically significant increases in RBC and
WBC counts compared to LPS and VE groups,
decreases in percentage of basophils and eosinophils
compared to control group, and decrease in percentage
of monocytes compared to control and LPS groups. PR
caused statistically significant increase in platelet counts
compared to LPS group, but this increases was in a
statistically significant way lower than that of control
level. It was stated that glucocorticoids increased RBC
and neutrophils, while they decreased lymphocytes,
basophils, and eosinophils in circulation (8). In the VE
administrated group, it was observed a statistically
significant decrease in percentage of eosinophils and
monocytes compared to the control group. In addition,
VE caused an increase of platelet counts compared to
LPS group, but this increase was under the level of
control value. It was found that VE inhibited platelet
aggregation in endotoxaemia and has antioxidant
activity. On the other hand, platelets may be aggregated
with hydrogen peroxide, and this aggregation may be
prevented by VE (21, 22).
In conclusion, LPS increased hepatic damage
markers, changed lipid metabolism and haematological
values. The administration of both PR and VE had a
protective effect against the endotoxaemic shock.
However, it may be assumed that PR was more effective
than VE in endotoxaemia.
Acknowledgments: This study was financed
by SUBAPK (2002/201).
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