Download Anna Moniuszko , Wojciech Łuczaj , Iwona Jarocka

Lipid peroxidation in Lyme neuroborreliosis
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Anna Moniuszko , Wojciech Łuczaj , Iwona Jarocka-Karpowicz , Sławomir Pancewicz ,
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Joanna Zajkowska , Katarzyna Bielawska , Elżbieta Skrzydlewska
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Department of Infectious Diseases and Neuroinfection, Medical University of Bialystok, Żurawia 14, 15-540 Bialystok, Poland
Department of Analytical Chemistry, Medical University of Bialystok, Mickiewicza 2d, 15-222 Bialystok, Poland
Section:
Models for decision making in economy or public health (tick-borne diseases)
Objectives:
Increase in number of Lyme disease (LD) patients is observed. In Europe, 85.000 new cases, and in North America 15-20.000 cases are noted yearly. Not fully clarified
LD pathogenesis and difficulties in diagnostics show necessity of searching indicators, which may let predict the course of the disease. Discovery of a biomarker of LD
should help in diagnostics. The main goal was to exam if neuroborreliosis (NB) effects on neuronal phospholipids peroxidation. We aimed to measure concentration of
PUFA’s cyclization and fragmentation products, antioxidants protecting PUFA’s against peroxidation – glutathione peroxidase and vitamin E in NB patients.
Methods:
22 patients with NB were included to the study. GSH-Px – EC.1.11.1.6 activity was assessed spectrophotometrically. Phospholipid arachidonic acid (AA) and
docosahexaenoic acid (DHA) were determined by gas chromatography. Lipid peroxidation was estimated by measuring of reactive aldehydes 4-hydroxynonenal (4HNE), 4-hydroxyhexenal (4-HHE), malondialdehyde (MDA), acrolein, crotonaldehyde, 4-oxononenal (4-ONE), F2-isoprostanes (8-isoPGF2α) and A4/J4neuroprostanes (NPs). Commercial assay kits were used to determine PLA2 and PAF-AH activities.
Table 2 The activity of glutathione peroxidase, phospholipase A2
and acetylhydrolase PAF and the level of vitamin E in the CSF
and plasma from patients with neuroborreliosis
Table 1. Results of blood laboratory tests of patients
with neuroborreliosis
a
b
Laboratory tests
– significantly different for p<0.001
- significantly different for p<0.005
NB patient, N= 22
CRP [mg/dl]
Control group
n=22
a
CSF GSH-Px
[mU/mg protein]
2.06 ± 0.82
1.71 ± 0.51
plasma GSH-Px
[mU/mg protein]
412 ± 79
307 ± 64
CSF vitamin E
[mg/L]
329 ± 113
269 ± 68
a
4.7 ± 0.45
plasma vitamin E
[mg/L]
10.67 ± 1.71
7.78 ± 2.44
a
256 ± 66
PLT [x100 000/ mm3]
AlAT [IU/ml]
CSF PLA2
nmoles/min/ml
26.8 ± 14.3
AspAT [IU/ml]
CSF PLA2
nmoles/min/ml
20.5 ± 5.7
plasma PLA2
[nmol/min/mL]
plasma PAF-AH
[nmol/min/mL]
5.11 ± 0.34
5.11 ± 0.14
9.31 ± 1.48
25.39 ± 5.58
4.75 ± 0.56
4.75 ± 0.16
5.56 ± 0.95
a
– significantly different for p<0.001
4-HHE
[pmol/mg creatinine]
MDA
[nmol/mg creatinine]
4-ONE
[pmol/mg creatinine]
3.96 ± 0.51
7.51 ± 0.93
1.30 ± 0.22
2.03 ± 0.52
0.04 ± 0.01
Neuroborreliosis
n=22
4.44 ± 2.80
24.88 ± 9.43
isoprostanes
[pg/ml]
68 ± 34
144 ± 42
neuroprostanes
[pg/ml]
383 ± 147
8416 ± 3451
4-HNE
[nmol/ml]
12.97 ± 4.52
25.72 ± 12.66a
4-HHE
[pmol/ml]
2.03 ± 1.01
3.32 ± 1.07a
MDA
[nmol/ml]
7.15 ± 3.72
27.34 ± 10.94a
4-ONE
[pmol/ml]
212 ± 62
309 ± 120b
phosholipid arachidonic acid
[ng/ml]
73 ± 10
66 ± 11
phospholipid docosahexaenoic acid
[ng/ml]
46 ± 6
40 ± 3
total isoprostanes
[ng/mL]
0.44 ± 0.17
4.05 ± 2.20a
free isoprostanes
[ng/mL]
0.16 ± 0.05
0.37 ± 0.19
neuroprostanes
[ng/mL]
0.67 ± 0.21
11.20 ± 4.90
MDA
[nmoL/mL]
1.37 ± 0.31
1.73 ± 0.27
a
4-HNE
[nmoL/mL]
0.12 ± 0.03
0.92 ± 0.21
a
4-HHE
[pmoL/mL]
6.96 ± 2.54
10.80 ± 2.91
acrolein
[pmoL/mL]
89 ± 24
221 ± 72
crotonaldehyde
[nmoL/mL]
0.64 ± 0.25
1.72 ± 0.49
4-ONE
[pmoL/mL]
74 ± 23
274 ± 62a
HNE-protein adducts
[pmol/g protein]
17.8 ± 6.4
23.4 ± 7.9d
phosholipid AA
[µg/mL]
71 ± 24
50 ± 13b
phospholipid LA
[µg/mL]
117 ± 31
57 ±19a
phosphlipid DHA
[µg/mL]
31 ± 8
21 ±7b
a
a
15.17 ± 2.04b
Table 4 The levels of lipid peroxidation products [8-isoprostanes
and MDA, 4-HNE, 4-HHE, 4-ONE] from patients with neuroborreliosis
Control group
n=22
Neuroborreliosis
n=22
CSF
8300 ± 3000
RBC [mln]
4-HNE
[nmol/mg creatinine]
Control group
n=19
Neuroborreliosis
n=22
24± 19
WBC [ /mm3]
8-isoprostanes
[ng/mg creatinine]
b
– significantly different for p<0.001; - significantly different for p<0.005
5.2 ± 6.3
SR [mm/h]
a
Table 3 The levels of phospholipid arachidonic acid and docosahexaenoic acid and lipid
peroxidation products [8-isoprostanes, neuroprostanes and MDA, 4-HNE, 4-HHE, 4-ONE]
in the cerebrospinal fluid and plasma from patients with neuroborreliosis
a
2.95 ± 1.84a
7.85 ± 3.73a
plasma
a
a
a
a
0.20 ± 0.13a
Results:
Decrease in the antioxidant abilities against lipid peroxidation in CSF and plasma was observed. The activity of GSH-Px and vitamin E concentration in both body fluids
decreased. Concentration of 8-isoPGF2α increased in the CSF, plasma and urine. The total concentration of 8-isoPGF2 was higher in CSF and plasma. Concentration
of plasma free 8-isoprostanes was twice enhanced and of neuroprostanes increased 20 times in CSF and 16 times in plasma. MDA, 4-HNE, 4-HHE and 4-ONE CSF
concentration was 1.5-4.0 times higher. The plasma 4-HNE and 4-ONE concentration was over 7 and 3 times higher respectively in NB. Concentration of plasma
4-HNE, and 4-HNE-His-protein adducts increased in plasma. The urine MDA, 4-HNE, 4-HHE and 4-ONE concentration increased by 4, 3, 2, 5 times respectively.
The CSF fatty acid concentration decreased by about 18% and 10% for DHA and AA respectively. Concentration of plasma AA and DHA was lower by 29% and 28%
respectively. Phospholipase A2 and PAF acetylhydrolase activities were lower in plasma and in CSF.
Conclusions:
Lipid peroxidation plays role in the pathogenesis of NB. Measurement of its products concentration or enzymes activity may help in diagnostic process of NB.