Prolidase Enzyme Activity in Stroke Patients

NCT ID: NCT03334968

Last Updated: 2017-11-09

Basic Information

• Recruitment Status: COMPLETED
• Total Enrollment: 74 participants
• Study Classification: OBSERVATIONAL
• Study Start Date: 2016-05-01
• Study Completion Date: 2017-05-01

Brief Summary

Stroke is a major cerebrovascular disease that causes significant burdens for human health and life, including high morbidity, mortality, and disability. Prolidase enzyme activity was found in various organs, such as the heart, brain, thymus, kidney, lung, pancreas, and spleen, and in plasma, leukocytes, erythrocytes, and dermal fibroblasts. An increase in collagen turnover is known to be correlated with increased prolidase enzyme activity. The aim of this study was to investigate whether SPA levels in AIS patients can be used as a potential diagnostic and prognostic marker. SPA levels were prospectively evaluated in 37 patients aged between 20 and 85 years who were admitted within 24 hours of the onset of AIS. The control group included 37 healthy volunteers of similar age without any disease.

Detailed Description

Ischemic stroke constitutes about 80-85% of all stroke cases and is caused by the interruption of cerebral blood flow due to a blood clot. Because of reactive oxygen species (ROS) production and its oxidative metabolite activity, the brain is highly sensitive to oxidative stress. This characteristic plays an important role in the pathogenesis of ischemic and hemorrhagic brain injuries .Prolidase, which is a cytosolic exopeptidase and a member of the matrix metalloproteinase (MMP) family, cleaves iminodipeptides from carboxy-terminal ends of proline or hydroxyproline and is actively involved in collagen metabolism. It has been shown that brain MMP activity is correlated with nutritive/oxidative stress and increases during reperfusion. Furthermore, in stroke patients, elevated serum MMP levels have been reported. Biomarkers that predict the outcome and occurrence of ischemic stroke are critically important for prevention and treatment .However, serum prolidase activity (SPA) has not been previously assessed in acute ischemic stroke (AIS) patients to our knowledge. Therefore, in this study, it's aimed to investigate whether SPA levels in AIS patients can be used as a potential diagnostic and prognostic marker.In the study, 37 patients aged between 20 and 85 years who were admitted within 24 hours of the onset of AIS were prospectively evaluated. The control group consisted of 37 healthy volunteers of similar age without any disease. A comprehensive physical examination was performed consisting of a neurological examination, blood biochemistry and blood count tests, electrocardiography, and a posterior-anterior chest X-ray for all patients. AIS patients underwent transthoracic echocardiography, multi-slice computed tomography (CT), and bilateral carotid-vertebral artery Doppler ultrasonography. National Institutes of Health Stroke Scale (NIHSS) scores, measured at 24 hours, 48 hours, and 28 days after stroke, were used to determine stroke severity.Ischemic stroke subtypes classification was done according to the Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria, as cardioembolism, large-artery atherosclerosis, small artery occlusion, stroke of other determined cause, or undetermined cause. The measurement method for SPA was defined by Myara et al. The optimized method of Özcan et al. was used. Prolidase activity was evaluated with a spectrophotometric method, by measuring the proline levels. Briefly, 500 μL pre-incubation solution (50 mmol/L Tris hydrochloride buffer at power of hydrogen (pH) 7.8, with 1 mmol/L endogenous antioxidant glutathione (GSH), 5 mmol/L manganese(II) chloride (MnCl2), and 0.1% Triton X-100) and 100 μL serum were mixed; this mixture was then pre-incubated for 3 h at 37°C. A 100-μL volume of pre-incubation serum was added to 100 μL 144 mmol/L Gly-Pro solution, and this mixture was incubated for 30 min at 37°C. After the incubation, 1 ml 0.45 mol/L Trichloroacetic acid solution was added quickly to the incubation tube, and the incubation reaction was stopped. This mixture was centrifuged at 1500 rpm for 5 min, and 500 μL supernatant was removed.

Conditions

Stroke, Ischemic; Prolidase Deficiency

Keywords

Stroke; prolidase; prognosis; biomarker

Study Design

• Observational Model Type: CASE_CONTROL
• Study Time Perspective: PROSPECTIVE

Study Groups

Patient group

Acute ischemic stroke patients

No interventions assigned to this group

Control group

Those served as control group

No interventions assigned to this group

Eligibility Criteria

Inclusion Criteria

• aged between 20 and 85
• admitted within 24 hours of the onset of acute ischemic stroke

Exclusion Criteria

• patients with heart disease (such as myocardial infarction or heart failure
• chronic obstructive pulmonary disease,
• pulmonary embolism,
• pulmonary hypertension,
• tuberculosis,
• lung cancer,
• chronic renal failure,
• current hormone replacement treatment.

• Minimum Eligible Age: 20 Years
• Maximum Eligible Age: 85 Years
• Eligible Sex: ALL
• Accepts Healthy Volunteers: Yes

Sponsors

Bezmialem Vakif University

OTHER

Sponsor Role: lead

Responsible Party

Abdulkadir Tunc

Neurologist

Responsibility Role: PRINCIPAL_INVESTIGATOR

Principal Investigators

Abdulkadir TUNÇ, MD

Role: PRINCIPAL_INVESTIGATOR

Bezmialem Vakif University

Locations

Bezmialem University

Istanbul, Fatih, Turkey (Türkiye)

Countries

Turkey (Türkiye)

References

Gonullu H, Aslan M, Karadas S, Kati C, Duran L, Milanlioglu A, Aydin MN, Demir H. Serum prolidase enzyme activity and oxidative stress levels in patients with acute hemorrhagic stroke. Scand J Clin Lab Invest. 2014 Apr;74(3):199-205. doi: 10.3109/00365513.2013.873949. Epub 2014 Jan 23.

Reference Type: RESULT

PMID: 24456419 (View on PubMed)

Verma AK, Raj J, Sharma V, Singh TB, Srivastava S, Srivastava R. Plasma Prolidase Activity and Oxidative Stress in Patients with Parkinson's Disease. Parkinsons Dis. 2015;2015:598028. doi: 10.1155/2015/598028. Epub 2015 Aug 9.

Reference Type: RESULT

PMID: 26347150 (View on PubMed)

Tabur S, Oguz E, Eren MA, Korkmaz H, Savas E, Aksoy N, Sabuncu T. Serum prolidase activity is associated with non-diabetic metabolic syndrome. Diabetol Metab Syndr. 2014 Dec 17;6(1):142. doi: 10.1186/1758-5996-6-142. eCollection 2014.

Reference Type: RESULT

PMID: 25540672 (View on PubMed)

Gasche Y, Copin JC, Sugawara T, Fujimura M, Chan PH. Matrix metalloproteinase inhibition prevents oxidative stress-associated blood-brain barrier disruption after transient focal cerebral ischemia. J Cereb Blood Flow Metab. 2001 Dec;21(12):1393-400. doi: 10.1097/00004647-200112000-00003.

Reference Type: RESULT

PMID: 11740200 (View on PubMed)

Myara I, Charpentier C, Lemonnier A. Optimal conditions for prolidase assay by proline colorimetric determination: application to iminodipeptiduria. Clin Chim Acta. 1982 Oct 27;125(2):193-205. doi: 10.1016/0009-8981(82)90196-6.

Reference Type: RESULT

PMID: 7139961 (View on PubMed)

CHINARD FP. Photometric estimation of proline and ornithine. J Biol Chem. 1952 Nov;199(1):91-5. No abstract available.

Reference Type: RESULT

PMID: 12999819 (View on PubMed)

Danton GH, Dietrich WD. Inflammatory mechanisms after ischemia and stroke. J Neuropathol Exp Neurol. 2003 Feb;62(2):127-36. doi: 10.1093/jnen/62.2.127.

Reference Type: RESULT

PMID: 12578222 (View on PubMed)

Other Identifiers

StrokeProlidase

Identifier Type: -

Identifier Source: org_study_id