Evaluation of Serum Levels of Irisin and Nesfatin-1 in Patients with Migraine without Aura

68 Int J Med Biochem graine and neuropeptides. One of the newly identified peptide hormones, irisin, is composed of 112 amino acids and weighs ~12 kDa [6, 7]. It should be noted that irisin affects autocrine, paracrine, and endocrine hormones. Several studies have found that irisin can be identified in the skeletal muscles, adipose tissue, kidneys, neural cells, ovaries, myocardial and smooth muscles, endothelium, liver, and pancreas [8, 9]. Also, recent observations have demonstrated that irisin has been detected at high levels in the brain, including the Purkinje cells of the cerebellum. Although the hormone irisin is mainly synthesized from muscle tissue, studies have shown the presence of irisin in the neuronal areas of rats [9, 10]. The physiological and hormonal effects of irisin still remain incompletely explained. Nesfatin-1 has a molecular weight of 9.7 kDa and is composed of 82 amino acids [11]. This newly discovered neuropeptide has antioxidant and antiapoptotic properties and plays a significant role in neurodegenerative damage and cerebrovascular events [12]. Nesfatin-1 has been found to be expressed mostly in the limbic system, the cortical nuclei, the dorsal motor nucleus of the vagus nerve, and the preganglionic sympathetic and parasympathetic neurons of the spinal cord [13]. One study concluded that nesfatin-1 plays a role in emotional and psychological events, such as panic disorder and chronic stress [14]. The present study is a preliminary assessment to determine serum irisin and nesfatin-1 levels in migraine without aura patients and examine whether these data could help with the identification or evaluation of the disease. Materials and Methods Eighty participants were included into the study in 2 groups: Migraine without aura patients (10 men, 30 women) and healthy subjects (10 men, 30 women). All of the participants were categorized in 10-year age groups from 20 to 59 years. The patients and healthy subjects had no metabolic diseases before the study. The exclusion criteria included any known history of headache disorder, endocrinopathy, cardiovascular disease, psychiatric disorder, cerebrovascular disease, neurological disorder, and the use of anti-inflammatory or steroid drugs, or anti-hypertensive medication. The patient data were obtained through self-reported questionnaires. The group of healthy controls was composed of participants with no migraine or headache complaints. Migraine without aura is the most frequent form of primary headache. Migraine without aura patients who were between migraine episodes were selected from individuals admitted to the neurology department of Konya Education and Research Hospital between March and December 2018. The migraine patients were diagnosed by an experienced neurologist according to the diagnostic criteria of the International Headache Society (IHS) (3rd edition, beta version). According to the IHS, the diagnostic criteria of migraine without aura are at least 5 headache attacks lasting 4-72 hours with/without treatment and at least 2 of the following a) pulsating quality, b) unilateral or bilateral location, c) pain intensity that is moderate to severe, and d) increased pain with physical activity (daily routine activity), as well as the experience of either vomiting and/or nausea, or phonophobia or photophobia during the headache. A visual analog scale (VAS) was used to assess pain intensity in the migraine without aura patients with a rating from 0 (no pain) to 10 (worst pain). Body mass index (BMI) was calculated as weight (kg) divided by the square of the height (m2) according to the WHO BMI classification (Technical Report Series 854). The duration of migraine without aura was determined by a single neurologist based on the patient questionnaires and clinical examination. Blood samples (10 mL) drawn from each patient were transferred to biochemistry tubes in order to evaluate the serum nesfatin-1 and irisin levels. The samples were separated after coagulation, centrifuged at 3000 rpm for 10 minutes, and stored in deep freeze at -800C until the run time. The serum levels of irisin and nesfatin-1 were measured using enzyme-linked immunosorbent assay (ELISA) kits. This study was approved by the ethics board of the Meram Faculty of Medicine of Necmettin Erbakan University in Konya, Turkey (approval number: 2018/1337-68) and conducted based on the principles of the Declaration of Helsinki. Written, informed consent was obtained from each participant prior to the study. Measurement of serum irisin and nesfatin-1 Serum irisin levels were measured using ELISA kits (Human Irisin ELISA Kit, catalog no: 201-12-5328; Shanghai Sunred Biological Technology, Shanghai, China). The analytical sensitivity, assay range, and intra-assay and inter-assay variation rates were 0.157 ng/mL, 0.2-60 ng/mL, <10%, and <12%, respectively. Serum nesfatin-1 levels were also measured using ELISA kits (Human NES 1 ELISA Kit, catalog no: 201-12-4341; Shanghai Sunred Biological Technology, Shanghai, China). The analytical sensitivity was 0.113 mmol/L, the assay range was 0.2-35 mmol/L, the intra-assay variation was < 10%, and the inter-assay variation was <12%. The kit manufacturer’s instructions were followed throughout all of the analyses. The absorbance of the samples was measured at 450 nm and recorded using a microplate reader (ELx800; BioTek Instruments, Winooski, VT, USA). Statistical analysis The statistical analyses were performed using IBM SPSS Statistics for Windows, Version 22.0 (IBM Corp., Armonk, NY, USA). Differences between the migraine without aura patients and the healthy controls were evaluated with the Mann-Whitney U test. The data were expressed as mean±SD (X±SD). One-way analysis of variance and the Kruskal-Wallis tests were used for age categories, VAS and BMI scores, and disease duration. The results were accepted as statistically significant at p<0.05. 69 Akdag, Irisin and nesfatin-1 in migraine / doi: 10.14744/ijmb.2020. 30301