The relationship of dysfunctional hdl, myeloperoxidase, lipid hydroperoxides levels with stroke violence and infarctus volume, and its prognostic value in determining disability in acute ischemic stroke

Introduction: Ischemic stroke is a neurological disorder that develops acutely as a result of decreased cerebral blood flow to a specific area of the brain due to vascular damage and is one of the most important causes of mortality and morbidity worldwide. Inflammatory mechanisms such as myeloperoxidase, superoxide free radicals, hydrogen peroxide, nitric oxide and lipid peroxidation, which lead to endothelial dysfunction and plaque instability, which are the basis of micro and macroangiopathy, which are formed as a result of increased oxidative stress in the ischemic stroke process, adversely affect anti-inflammatory and antiatherogenic functions, It initiates the atherogenic and thrombogenic process, which affects both the cardiovascular system and the cerebrovascular system, creating an important risk factor for stroke. The aim of this study is to examine the interaction of serum dysfunctional HDL, myeloperoxidase and lipid hydroperoxidase levels with clinical severity of stroke and infarct volume level in patients with acute ischemic stroke and to determine the prognostic value of these parameters in determining disability. Materials and Methods:This prospective descriptive case series consists of 100 patients aged 18-80 years who were admitted to Ankara City Hospital Emergency Department between April 2020 and September 2020 with acute ischemic stroke in the first 24 hours and were inpatient in the Neurology Clinic. It was carried out by obtaining written consents of 50 volunteer control group participants between the ages of 18-80 who applied to the Neurology Outpatient Clinic. In the clinical evaluation of the patients; early disability was detected in each patient using the National Institues Of Healt Stroke Scale Scores(NIHSS) and the Modified Rankin Scale(MRS) at discharge. In line with the information obtained from the patients history and medical history, the type of ischemic stroke, the presence of a previous stroke or sequelae, the drugs used including antiaggregant and anticoagulant drugs before and additional chronic diseases were noted. Immediately after the diagnosis of ischemic cerebrovascular disease is confirmed with clinical and cranial imaging, blood samples were collected from each participant to measure serum dysfunctional HDL, myeloperoxidase (MPO), lipid hydroperoxidase (LOOH), HBA1C, serum lipid profile ((total cholesterol (TC), LDL (Low density Lipoprotein), HDL (High density lipoprotein)) triglyceride (TG)). Biochemical evaluation of Serum Dysfunctional HDL (Paraxonase (PON) activity), serum MPO, serum LOOH was carried out by Ankara City Hospital Biochemistry Clinic. For the measurement of dysfunctional HDL, isolation of HDLs was performed based on the principle of high positive correlation between the dysfunctional feature of HDL and the myeloperoxidase / paraoxonase ratio and detecting the dysfunction of HDL. PON enzyme activity measurement was performed on the basis of spectrophotometric measurement of 4-nitrophenol formed as a result of enzymatic hydrolysis of paraoxon used as substrate at 412 nm wavelength. Paraoxon hydrolysis rates were evaluated by measuring 4-nitrophenol formation spectrophotometrically at 25C 412 nanometers. PON enzyme activity was used as a molar extinction coefficient of 17, 1000 M-1 cm. After taking samples for MPO measurement, it was waited for 30 minutes to complete coagulation, followed by centrifugation at 1300xg for 10 minutes. Samples were kept in each eppendorf tubes at -80 ° C until analysis. Measurement of MPO levels was made using commercially available Human Myeloperoxidase ELISA kits (Cayman Chemical, Michigan, USA). For the Human Myeloperoxidase ELISA kit, the detection range in the analysis of the samples was determined as 0.2-10 ng / mL and the sensitivity as 0.2 ng / mL. The within-run and inter-run CV (coefficient of variation) for the kit was determined as 3.83% and 8.13%, respectively. LOOH was measured by the xinelol orange method. The method was applied according to the principle that Fe + 2 oxidized to Fe + 3 with LOHH in an acidic environment. In order to calculate the infarct volume, the area of​the infarct was calculated with the Free hand ROI tool in DWI sections, and this area information obtained from each section was collected. The volume information was created by multiplying the section thickness of 5 mm by the sum of the interslice gap 1 mm (6 mm).Volume information was created by multiplying. In parallel with the infarct volume examination in patients with acute stroke, the statistical correlation of the relevant biochemical parameters was investigated. SPPS 25 (IBM Corp. Released 2017. IBM SPSS Statistics for Windows, Version 25.0. Armonk, NY: IBM Corp.) statistical package program was used to evaluate the data. The cut-off scores of the patients and healthy individuals according to the measurement parameters were evaluated by ROC analysis. AUC Value, Sensitivity, Selectivity values were calculated. A p<0.05 level was considered statistically significant. Results: Patient group was evaluated with demographic data (ie age, gender, race, marital status, chronic diseases and drug use), clinical disability scores (arrival NIHS, discharge NIHSS, discharge mRS), MRI infarct volume (ROI CM2) and serum HBA1C, TC, LDL , TG, HDL, LOOH, MPO and MPO / PON. The control group was evaluated with serum TC, HDL, LDL, TG, LOOH, MPO and MPO-PON. 57% of the patients included in the study were female (n: 57) 48% (n: 48) were male; 52% of the control group was female (n: 26) and 48% were male (n: 24). The mean age was 66.31 ± 11.947 in the patient group and 64.9 ± 6.11 in the control group. Gender distribution and mean ages were similar between the groups (p =0.561; p = 0.121, respectively). Serum TC, HDL, LDL, TG, PON, LOOH, MPO and MPO / PON values ​​were measured and compared in the patient and control groups. Except for TG, other measured parameters showed statistically significant difference between groups. While TC, LDL, LOOH, MPO levels and MPO / PON ratio were lower in healthy controls than in the patient group; HDL and PON were found to be significantly lower in the patient group, although the TG level was found to be high in patients with stroke, it did not reach statistical significance. Correlation analyzes were performed to investigate whether there was a relationship between serum TC, HDL, LDL, TG, PON, LOOH, MPO, MPO / PON levels and arrival NIHSS, discharge NIHSS, discharge MRS and infarct volume (ROI (cm 3). A weak negative correlation was found between NIHSS and between LOOH and discharge mRS (r =-0.204 p<0.05; r = -0.244 p<0.05,). Logistic regression analyzes performed to reveal factors that may affect stroke severity (NIHSS score 6 and above) and early disability (mRS score 2 and above) did not show statistical significance for lipid profile, LOOH, PON, MPO, MPO / PON. Hypertension (OR =3.287, 95% CI = 1.080-10.009, p = 0.036), presence of diabetes (OR = 0.292, 95% CI = 0.089-0.962, p = 0.043) and HbA1c levels (OR = 1.487, 95) % CI = 1.096- 2.019, p = 0.011) were determined as an independent risk factor for stroke severity. For early disability, presence of diabetes (OR = 0.011, 95% CI = 0.000-0.893, p = 0.044), HbA1c levels (OR = 5.059, 95% CI = 1.222-20.939, p = 0.025), infarct volume (OR = 1.027, 95% CI = 1.008-1.047, p = 0.005) and arrival NIHSS score (OR = 21.646, 95% CI = 3.113-150.524, p = 0.002) were determined as independent risk factors. ROC curve analyzes performed to reveal the sensitivity and specificity of lipid profile, PON, MPO, LOOH levels and PON / MPO ratio in predicting stroke severity, and early disability revealed statistical significance only for LOOH. The cut-off point in the mRS score for early disability is accepted as 2, LOOH, with 68.49% sensitivity and 55.56% selectivity (p = 0.0375); The cut-off point is accepted as 4 in the mRS score, it can predict disability with 76.00% sensitivity and 57.33% selectivity (p = 0.0377). The cut off value is 7.2 and 6.2, respectively. Conclusion: Conclusion: In our study, serum dysfunctional HDL, which is an important atherosclerotic risk factor contributing to the inflammatory process in patients with acute stroke, MPO, which is a lysosomal enzyme secreted from leukocytes in response to oxidative stress and is an important etiological factor in cerebrovascular events by causing endothelial dysfunction and PON showing its activity, LOOH activity levels, which are indicators of increased levels of free oxygen radicals due to the effect of MPO / PON values ​​and cerebral blood flow, relationship with stroke severity and infarct volume, and its prognostic value in determining disability were examined. It has been shown that these markers are significantly increased in serum levels in the patient group and it has been statistically proven that they contribute to the infarct volume and clinical disability. Although the relationship between low HDL levels and recurrence of stroke has been demonstrated in studies up to now, there are not enough studies on the effect of HDL functionality in stroke patients, rather than HDL level. In this respect, our study will be seminal in terms of showing the correlation between dysfunctional HDL and infarct volume, as well as PON activity and stroke severity in stroke patients. As such, dysfunctional HDL should be included in lipid profile test batteries when evaluating stroke patients. Many studies with similar predictions are needed in this area.