- Email: [email protected]
Dynamic changes in calprotectin and its correlation with traditional markers of oxidative stress in patients with acute ischemic stroke
Accepted Manuscript Dynamic changes of calprotectin and its correlation with traditional markers of oxidative stress in patients with acute ischemic stroke Antonios Chatzopoulos, Aspasia I. Tzani, Ilias P. Doulamis, Panagiotis S. Konstantopoulos, Dionysia Birba, Christos Verikokos, Nikolaos Tentolouris, Gavriil Karatzas, Despina N. Perrea PII:
S1109-9666(17)30116-1
DOI:
10.1016/j.hjc.2017.07.002
Reference:
HJC 184
To appear in:
Hellenic Journal of Cardiology
Received Date: 9 March 2017 Revised Date:
30 June 2017
Accepted Date: 3 July 2017
Please cite this article as: Chatzopoulos A, Tzani AI, Doulamis IP, Konstantopoulos PS, Birba D, Verikokos C, Tentolouris N, Karatzas G, Perrea DN, Dynamic changes of calprotectin and its correlation with traditional markers of oxidative stress in patients with acute ischemic stroke, Hellenic Journal of Cardiology (2017), doi: 10.1016/j.hjc.2017.07.002. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Dynamic changes of calprotectin and its correlation with traditional markers of oxidative stress in patients with acute ischemic stroke
RI PT
Antonios Chatzopoulos1+, Aspasia I. Tzani1+, Ilias P. Doulamis1, Panagiotis S. Konstantopoulos1, Dionysia Birba1, Christos Verikokos2, Nikolaos Tentolouris3, Gavriil
SC
Karatzas2, Despina N. Perrea1
These authors contributed equally
1
Laboratory for Experimental Surgery and Surgical Research “N.S Christeas”, Medical School,
M AN U
+
National and Kapodistrian University of Athens, Athens, Greece 2
Second Department of Propedeutic Surgery, 'Laiko' General Hospital, Medical School,
National and Kapodistrian University of Athens, Athens, Greece
First Department of Propaedeutic and Internal Medicine, 'Laiko' General Hospital, Medical
TE D
3
School, National and Kapodistrian University of Athens, Athens, Greece.
EP
Correspondence:
AC C
Aspasia Tzani, M.D, Ph.D.c
Laboratory for Experimental Surgery and Surgical Research “N.S Christeas”, Medical School, National and Kapodistrian University of Athens, Athens, Greece Agiou Thoma 15b, Athens,Greece, 11527 Email: [email protected]
ACCEPTED MANUSCRIPT
Acute ischemic stroke (AIS) is a medical condition rising from the obstruction of blood flow to cerebral circulation for more than 30 minutes, leading thus to brain tissue damage.[1, 2] AIS
RI PT
triggers a pro-inflammatory cascade both on a local and on a systemic level.[3] Calprotectin is a relatively new marker indicating inflammatory process and it has been thoroughly studied in carcinogenesis. [4, 5]
SC
We sought to investigate for the first time the alternations of “traditional” biomarkers of oxidative stress and a novel marker –namely calprotectin- for inflammation shortly and mid-term
M AN U
after AIS and to elucidate their potent correlation with characteristics from patients’ history. After the hospital ethics committee approval, patients, who were admitted within one hour to the General Hospital of Argos, Emergency Department between September 2015 and 2016 with acute stroke symptoms were evaluated. Forty-two patients, which were diagnosed as having AIS
TE D
based on medical history, neurological examination findings and brain CT scans, were included. Information about the study was provided to patients and their consents were obtained. Because oxidant levels can change, patients with a history of cerebrovascular disease, neurological
EP
disease, chronic kidney disease, blood disorders and a history of stroke attack were excluded. Participants involved in our study as controls were healthy blood donors and were without any
AC C
acute or chronic diseases.
To assess dynamic changes of nitric oxide (NO) and calprotectin, EDTA venous blood samples were collected within the first hour of their hospital admission (t0) and after 8 hours (t1).NO plays a predominant role in this procedure, initially as protective factor in the early stages following cerebral ischemia (< 2h) and with adverse effects afterwards (> 6h).[6] Moreover,
ACCEPTED MANUSCRIPT
current data suggest fluctuation of calprotectin levels between these time points post-ischemia .[7] Serum Hydrogen Peroxide (PerOx), NO and calprotectin levels were measured by enzyme-
RI PT
linked immunosorbent assay (ELISA) using commercial assay kits. Because of the NO
instability, the quantitative of NO was indirectly determined based on the detection and the calculation of the nitrite to nitrate molar ratio.
SC
Statistical Analysis was performed using SPSS 23.0 (Windows Version). Paired T-test and Mann–Whitney U test were used accordingly. All performed tests were two sided. A p value less
M AN U
than 0.05 was considered statistically significant (p = 0.05).
42 patients (mean age 72.67 ± 7.851 years)(39 males and 13 females) with AIS as well as 23 age-matched healthy control individuals ( mean age70.91 ± 9.01 years) were included. Mean body mass index(BMI) of the patients was 28.9 (range 20.9- 40.4). 64% of the patients were
TE D
hypertensive, while 38% and 23% of the patients had a history of diabetes mellitus and coronary artery disease, respectively. Only five patients had documented hyperlipidemia and four patients had atrial fibrillation. Three patients did not have any comorbidity.
EP
Baseline serum levels of NO as Nitrate/Nitrite concentration were lower in patients group than control group (-1.870 ± 1,820 vs 0.034 ± 0.048, p<0.001). (Figure 1)
AC C
After 8 hours serum levels of NO (0.949 ± 2.191) were higher than baseline (-1.870 ± 1.820) in the patients group ( p<0.05). Additionally, serum calprotectin levels were higher at baseline than t1 in the patients group (12.983 ± 1.377 vs 12.363 ± 0.990 mmol/l, p<0.01). (Figure 1) When dividing the patients in two subgroups with reference to their blood pressure, it was found that hypertensive patients(64.3%) had lower levels of NO at baseline compared with non-
ACCEPTED MANUSCRIPT
hypertensive patients (-2.387 ± 1.739 vs -0.938 ± 1.624, p = 0.012); this difference was not observed after 8 hours. Moreover, subdivision of the patients into two groups regarding the presence of coronary artery
RI PT
disease(CAD) revealed that patients with CAD(24%) had higher serum levels of NO at t0 and lower serum levels of PerOx at t1 compared with patients without CAD (p = 0.003 and p = 0.001, respectively).
SC
Categorization of the patients based on their Diabetes Mellitus (DM) history illustrated that serum calprotectin levels in diabetic patients(38%) were lower than non-diabetic patients at
M AN U
baseline (12.215 ± 1.363 vs 13.481 ± 1.170 mmol/l, p = 0.014). This difference was not observed after 8 hours.
In terms of pairwise associations, significant positive and negative correlations were found between measured parameters in patients and control group (Table 1).
TE D
In the present study, the effect of AIS on oxidative stress markers was investigated. Patients with AIS had lower NO serum levels than healthy age-matched subjects at their hospital admission, whereas no significant difference was observed regarding PerOx levels. Eight hours
EP
after the hospital admission, NO levels were increased, while calprotectin serum levels were lower than initial levels. Furthermore, hypertensive patients had lower serum NO levels at
AC C
baseline compared with non-hypertensive, while patients with CAD had higher levels of NO at baseline compared with non-CAD patients. Our main finding is the implication of calprotectin in AIS. The involvement of calprotectin in the pathogenesis of cardiovascular disease has attracted increasing amount of interest.[8] In humans, calprotectin correlates with the progress of coronary and carotid atherosclerosis and with vulnerable plaque phenotype.[9] We are the first, to our knowledge, to propose a correlation
ACCEPTED MANUSCRIPT
between serum calprotectin levels and AIS. According to our findings, calprotectin levels were higher in baseline than 8 h after the incidence. This observation depicts an acute response of macrophages, which predominantly release calprotectin, to brain tissue ischemia.[10] Taking
calprotectin exerts a role in regulation of NO.
RI PT
into account the negative correlation between calprotectin and NO levels, it can be assumed that
The increase of calprotectin is immediate and short-term, since it returns to lower levels
SC
after few hours. The clinical implication of these characteristics has two meaningful aspects; on the one hand calprotectin can be used as diagnostic biomarker for AIS and on the other hand,
M AN U
thanks to its short duration increase, it can act as marker for reinfarction of cerebral blood vessels.
This study depicts the dynamic changes in the oxidative status at the early stages after AIS and proposes calprotectin as a novel serum biomarker for AIS. This association ferments ground for
EP
Acknowledgements
TE D
new implications both in clinical practice and diagnostic techniques.
data.
AC C
We wish to thank Mrs Pelagia Kyriazopoulou for her kind assistance in statistical analysis of our
Funding
This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.
Declaration of conflicting interests The Authors declare that there is no conflict of interest.
M AN U
SC
RI PT
ACCEPTED MANUSCRIPT
References
4. 5. 6. 7. 8.
9.
10.
TE D
3.
EP
2.
Akoumianakis, I.D. and C.A. Antoniades, Studying Systemic Oxidative Stress in Heart Failure: Does It Have Any Role in Clinical Practice? Hellenic J Cardiol, 2015. 56(5): p. 402-5. Stefanadis, C.I., Risk stratification for stroke: do we need new tools? Hellenic J Cardiol, 2013. 54(5): p. 417. Benetos, G., et al., Bilateral symmetry of local inflammatory activation in human carotid atherosclerotic plaques. Hellenic J Cardiol, 2015. 56(2): p. 118-24. Tabur, S., et al., Serum calprotectin: a new potential biomarker for thyroid papillary carcinoma. Tumour Biol, 2015. 36(10): p. 7549-56. Striz, I. and I. Trebichavsky, Calprotectin - a pleiotropic molecule in acute and chronic inflammation. Physiol Res, 2004. 53(3): p. 245-53. Moro, M.A., et al., Role of nitric oxide after brain ischaemia. Cell Calcium, 2004. 36(3-4): p. 26575. Ebbing, J., et al., Dynamics of Urinary Calprotectin after Renal Ischaemia. PLoS One, 2016. 11(1): p. e0146395. Pedersen, L., et al., Plasma calprotectin and its association with cardiovascular disease manifestations, obesity and the metabolic syndrome in type 2 diabetes mellitus patients. BMC Cardiovasc Disord, 2014. 14: p. 196. Santilli, F., et al., Circulating myeloid-related protein-8/14 is related to thromboxane-dependent platelet activation in patients with acute coronary syndrome, with and without ongoing lowdose aspirin treatment. J Am Heart Assoc, 2014. 3(4). Dessing, M.C., et al., The calcium-binding protein complex S100A8/A9 has a crucial role in controlling macrophage-mediated renal repair following ischemia/reperfusion. Kidney Int, 2015. 87(1): p. 85-94.
AC C
1.
ACCEPTED MANUSCRIPT
Table 1: Spearman correlations among measured parameters in patients group and in patients subgroups.
Patients group
Calprotectin
t0
NO
t1
Calprotectin-t1 Calprotectin-t0
PerOX
NO
Calprotectin
- 0.472
0.011
0.033
Calprotectin-t1
0.541
0.017
t0
Calprotectin-t0
- 0.505
0.027
t1
Calprotectin-t1
- 0.606
0.006
t0
Calprotectin-t1
0.650
0.003
NO-t1
0.749
0.013
Calprotectin-t1
0.627
0.039
t0
t0
AC C
Patients with DM
0.035
- 0.411
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Patients with CAD
-0.399
NO-t0
NO
Calprotectin
< 0,001
EP
Patients with HTN
0.718
M AN U
Calprotectin-t1
p
RI PT
r
SC
Time point
t0
Spearman correlation coefficients (??) are indicated; ?? < 0.05 is considered statistically significant. t0: within 1st hour after hospital admission; t1: 8 hours after hospital admission. NO: Nitric Oxide; PerOx: Hydrogen Peroxide; HTN: Hypertension; CAD: Coronary Artery Disease; DM: Diabetes Mellitus
ACCEPTED MANUSCRIPT
Figure Legend
in patients with acute ischemic stroke and healthy controls
RI PT
Figure 1. Dynamic changes of serum concentrations of oxidative stress markers and calprotectin
A: PerOx (µmoL/L) serum levels between patients during admission and healthy controls;
SC
B:NO (Nitrate/Nitrite concentration) serum levels between patients during admission and healthy controls;
M AN U
C:PerOx (µmoL/L) levels in patients’ serum between t0 and 8 h after the ischemic attack; D: NO (Nitrate/Nitrite concentration) levels in patients’ serum between t0 and 8 h after the ischemic attack;
D:Calprotectin(mmol/l) levels in patients’ serum between t0 and 8 h after the ischemic attack.
AC C
EP
TE D
PerOx: Hydrogen Peroxide; NO: Nitric Oxide
AC C
EP
TE D
M AN U
SC
RI PT
ACCEPTED MANUSCRIPT
S1109-9666(17)30116-1
DOI:
10.1016/j.hjc.2017.07.002
Reference:
HJC 184
To appear in:
Hellenic Journal of Cardiology
Received Date: 9 March 2017 Revised Date:
30 June 2017
Accepted Date: 3 July 2017
Please cite this article as: Chatzopoulos A, Tzani AI, Doulamis IP, Konstantopoulos PS, Birba D, Verikokos C, Tentolouris N, Karatzas G, Perrea DN, Dynamic changes of calprotectin and its correlation with traditional markers of oxidative stress in patients with acute ischemic stroke, Hellenic Journal of Cardiology (2017), doi: 10.1016/j.hjc.2017.07.002. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Dynamic changes of calprotectin and its correlation with traditional markers of oxidative stress in patients with acute ischemic stroke
RI PT
Antonios Chatzopoulos1+, Aspasia I. Tzani1+, Ilias P. Doulamis1, Panagiotis S. Konstantopoulos1, Dionysia Birba1, Christos Verikokos2, Nikolaos Tentolouris3, Gavriil
SC
Karatzas2, Despina N. Perrea1
These authors contributed equally
1
Laboratory for Experimental Surgery and Surgical Research “N.S Christeas”, Medical School,
M AN U
+
National and Kapodistrian University of Athens, Athens, Greece 2
Second Department of Propedeutic Surgery, 'Laiko' General Hospital, Medical School,
National and Kapodistrian University of Athens, Athens, Greece
First Department of Propaedeutic and Internal Medicine, 'Laiko' General Hospital, Medical
TE D
3
School, National and Kapodistrian University of Athens, Athens, Greece.
EP
Correspondence:
AC C
Aspasia Tzani, M.D, Ph.D.c
Laboratory for Experimental Surgery and Surgical Research “N.S Christeas”, Medical School, National and Kapodistrian University of Athens, Athens, Greece Agiou Thoma 15b, Athens,Greece, 11527 Email: [email protected]
ACCEPTED MANUSCRIPT
Acute ischemic stroke (AIS) is a medical condition rising from the obstruction of blood flow to cerebral circulation for more than 30 minutes, leading thus to brain tissue damage.[1, 2] AIS
RI PT
triggers a pro-inflammatory cascade both on a local and on a systemic level.[3] Calprotectin is a relatively new marker indicating inflammatory process and it has been thoroughly studied in carcinogenesis. [4, 5]
SC
We sought to investigate for the first time the alternations of “traditional” biomarkers of oxidative stress and a novel marker –namely calprotectin- for inflammation shortly and mid-term
M AN U
after AIS and to elucidate their potent correlation with characteristics from patients’ history. After the hospital ethics committee approval, patients, who were admitted within one hour to the General Hospital of Argos, Emergency Department between September 2015 and 2016 with acute stroke symptoms were evaluated. Forty-two patients, which were diagnosed as having AIS
TE D
based on medical history, neurological examination findings and brain CT scans, were included. Information about the study was provided to patients and their consents were obtained. Because oxidant levels can change, patients with a history of cerebrovascular disease, neurological
EP
disease, chronic kidney disease, blood disorders and a history of stroke attack were excluded. Participants involved in our study as controls were healthy blood donors and were without any
AC C
acute or chronic diseases.
To assess dynamic changes of nitric oxide (NO) and calprotectin, EDTA venous blood samples were collected within the first hour of their hospital admission (t0) and after 8 hours (t1).NO plays a predominant role in this procedure, initially as protective factor in the early stages following cerebral ischemia (< 2h) and with adverse effects afterwards (> 6h).[6] Moreover,
ACCEPTED MANUSCRIPT
current data suggest fluctuation of calprotectin levels between these time points post-ischemia .[7] Serum Hydrogen Peroxide (PerOx), NO and calprotectin levels were measured by enzyme-
RI PT
linked immunosorbent assay (ELISA) using commercial assay kits. Because of the NO
instability, the quantitative of NO was indirectly determined based on the detection and the calculation of the nitrite to nitrate molar ratio.
SC
Statistical Analysis was performed using SPSS 23.0 (Windows Version). Paired T-test and Mann–Whitney U test were used accordingly. All performed tests were two sided. A p value less
M AN U
than 0.05 was considered statistically significant (p = 0.05).
42 patients (mean age 72.67 ± 7.851 years)(39 males and 13 females) with AIS as well as 23 age-matched healthy control individuals ( mean age70.91 ± 9.01 years) were included. Mean body mass index(BMI) of the patients was 28.9 (range 20.9- 40.4). 64% of the patients were
TE D
hypertensive, while 38% and 23% of the patients had a history of diabetes mellitus and coronary artery disease, respectively. Only five patients had documented hyperlipidemia and four patients had atrial fibrillation. Three patients did not have any comorbidity.
EP
Baseline serum levels of NO as Nitrate/Nitrite concentration were lower in patients group than control group (-1.870 ± 1,820 vs 0.034 ± 0.048, p<0.001). (Figure 1)
AC C
After 8 hours serum levels of NO (0.949 ± 2.191) were higher than baseline (-1.870 ± 1.820) in the patients group ( p<0.05). Additionally, serum calprotectin levels were higher at baseline than t1 in the patients group (12.983 ± 1.377 vs 12.363 ± 0.990 mmol/l, p<0.01). (Figure 1) When dividing the patients in two subgroups with reference to their blood pressure, it was found that hypertensive patients(64.3%) had lower levels of NO at baseline compared with non-
ACCEPTED MANUSCRIPT
hypertensive patients (-2.387 ± 1.739 vs -0.938 ± 1.624, p = 0.012); this difference was not observed after 8 hours. Moreover, subdivision of the patients into two groups regarding the presence of coronary artery
RI PT
disease(CAD) revealed that patients with CAD(24%) had higher serum levels of NO at t0 and lower serum levels of PerOx at t1 compared with patients without CAD (p = 0.003 and p = 0.001, respectively).
SC
Categorization of the patients based on their Diabetes Mellitus (DM) history illustrated that serum calprotectin levels in diabetic patients(38%) were lower than non-diabetic patients at
M AN U
baseline (12.215 ± 1.363 vs 13.481 ± 1.170 mmol/l, p = 0.014). This difference was not observed after 8 hours.
In terms of pairwise associations, significant positive and negative correlations were found between measured parameters in patients and control group (Table 1).
TE D
In the present study, the effect of AIS on oxidative stress markers was investigated. Patients with AIS had lower NO serum levels than healthy age-matched subjects at their hospital admission, whereas no significant difference was observed regarding PerOx levels. Eight hours
EP
after the hospital admission, NO levels were increased, while calprotectin serum levels were lower than initial levels. Furthermore, hypertensive patients had lower serum NO levels at
AC C
baseline compared with non-hypertensive, while patients with CAD had higher levels of NO at baseline compared with non-CAD patients. Our main finding is the implication of calprotectin in AIS. The involvement of calprotectin in the pathogenesis of cardiovascular disease has attracted increasing amount of interest.[8] In humans, calprotectin correlates with the progress of coronary and carotid atherosclerosis and with vulnerable plaque phenotype.[9] We are the first, to our knowledge, to propose a correlation
ACCEPTED MANUSCRIPT
between serum calprotectin levels and AIS. According to our findings, calprotectin levels were higher in baseline than 8 h after the incidence. This observation depicts an acute response of macrophages, which predominantly release calprotectin, to brain tissue ischemia.[10] Taking
calprotectin exerts a role in regulation of NO.
RI PT
into account the negative correlation between calprotectin and NO levels, it can be assumed that
The increase of calprotectin is immediate and short-term, since it returns to lower levels
SC
after few hours. The clinical implication of these characteristics has two meaningful aspects; on the one hand calprotectin can be used as diagnostic biomarker for AIS and on the other hand,
M AN U
thanks to its short duration increase, it can act as marker for reinfarction of cerebral blood vessels.
This study depicts the dynamic changes in the oxidative status at the early stages after AIS and proposes calprotectin as a novel serum biomarker for AIS. This association ferments ground for
EP
Acknowledgements
TE D
new implications both in clinical practice and diagnostic techniques.
data.
AC C
We wish to thank Mrs Pelagia Kyriazopoulou for her kind assistance in statistical analysis of our
Funding
This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.
Declaration of conflicting interests The Authors declare that there is no conflict of interest.
M AN U
SC
RI PT
ACCEPTED MANUSCRIPT
References
4. 5. 6. 7. 8.
9.
10.
TE D
3.
EP
2.
Akoumianakis, I.D. and C.A. Antoniades, Studying Systemic Oxidative Stress in Heart Failure: Does It Have Any Role in Clinical Practice? Hellenic J Cardiol, 2015. 56(5): p. 402-5. Stefanadis, C.I., Risk stratification for stroke: do we need new tools? Hellenic J Cardiol, 2013. 54(5): p. 417. Benetos, G., et al., Bilateral symmetry of local inflammatory activation in human carotid atherosclerotic plaques. Hellenic J Cardiol, 2015. 56(2): p. 118-24. Tabur, S., et al., Serum calprotectin: a new potential biomarker for thyroid papillary carcinoma. Tumour Biol, 2015. 36(10): p. 7549-56. Striz, I. and I. Trebichavsky, Calprotectin - a pleiotropic molecule in acute and chronic inflammation. Physiol Res, 2004. 53(3): p. 245-53. Moro, M.A., et al., Role of nitric oxide after brain ischaemia. Cell Calcium, 2004. 36(3-4): p. 26575. Ebbing, J., et al., Dynamics of Urinary Calprotectin after Renal Ischaemia. PLoS One, 2016. 11(1): p. e0146395. Pedersen, L., et al., Plasma calprotectin and its association with cardiovascular disease manifestations, obesity and the metabolic syndrome in type 2 diabetes mellitus patients. BMC Cardiovasc Disord, 2014. 14: p. 196. Santilli, F., et al., Circulating myeloid-related protein-8/14 is related to thromboxane-dependent platelet activation in patients with acute coronary syndrome, with and without ongoing lowdose aspirin treatment. J Am Heart Assoc, 2014. 3(4). Dessing, M.C., et al., The calcium-binding protein complex S100A8/A9 has a crucial role in controlling macrophage-mediated renal repair following ischemia/reperfusion. Kidney Int, 2015. 87(1): p. 85-94.
AC C
1.
ACCEPTED MANUSCRIPT
Table 1: Spearman correlations among measured parameters in patients group and in patients subgroups.
Patients group
Calprotectin
t0
NO
t1
Calprotectin-t1 Calprotectin-t0
PerOX
NO
Calprotectin
- 0.472
0.011
0.033
Calprotectin-t1
0.541
0.017
t0
Calprotectin-t0
- 0.505
0.027
t1
Calprotectin-t1
- 0.606
0.006
t0
Calprotectin-t1
0.650
0.003
NO-t1
0.749
0.013
Calprotectin-t1
0.627
0.039
t0
t0
AC C
Patients with DM
0.035
- 0.411
TE D
Patients with CAD
-0.399
NO-t0
NO
Calprotectin
< 0,001
EP
Patients with HTN
0.718
M AN U
Calprotectin-t1
p
RI PT
r
SC
Time point
t0
Spearman correlation coefficients (??) are indicated; ?? < 0.05 is considered statistically significant. t0: within 1st hour after hospital admission; t1: 8 hours after hospital admission. NO: Nitric Oxide; PerOx: Hydrogen Peroxide; HTN: Hypertension; CAD: Coronary Artery Disease; DM: Diabetes Mellitus
ACCEPTED MANUSCRIPT
Figure Legend
in patients with acute ischemic stroke and healthy controls
RI PT
Figure 1. Dynamic changes of serum concentrations of oxidative stress markers and calprotectin
A: PerOx (µmoL/L) serum levels between patients during admission and healthy controls;
SC
B:NO (Nitrate/Nitrite concentration) serum levels between patients during admission and healthy controls;
M AN U
C:PerOx (µmoL/L) levels in patients’ serum between t0 and 8 h after the ischemic attack; D: NO (Nitrate/Nitrite concentration) levels in patients’ serum between t0 and 8 h after the ischemic attack;
D:Calprotectin(mmol/l) levels in patients’ serum between t0 and 8 h after the ischemic attack.
AC C
EP
TE D
PerOx: Hydrogen Peroxide; NO: Nitric Oxide
AC C
EP
TE D
M AN U
SC
RI PT
ACCEPTED MANUSCRIPT