Josip Juraj Strossmayer University of Osijek. Faculty of Medicine.
Abstract
Uvod: Migrene predstavljaju jednu od vodećih bolesti današnjice. Najveći problem, upravo je dijagnostika migrena. Ispitujemo povezanost dva biljega oštećenja moždanog tkiva (S100B i NSE) s migrenom. Ciljevi istraživanja: Utvrditi ekspresiju navedenih biljega i povezanost njihove ekspresije s višegodišnjim osobitostima kliničke slike pacijenata s migrenskom glavoboljom. Materijali i metode: Uzorke venske krvi uzorkovani su od osoba koje smo podijelili u dvije skupine - skupinu ispitanika i kontrolnu skupinu. Uključni kriteriji za skupinu ispitanika: višegodišnja dijagnoza migrenske glavobolje, manje od četrdeset godina te krv uzorkovana u roku dva sata od početka napada. Kriterij za isključenje je dob iznad 40 godina te postojanje drugih organskih i psihijatrijskih bolesti, izuzev depresije. Kontrolnu skupinu čine ispitanici bez glavobolje niti bilo kakve druge ozbiljne bolesti, a uzorci im se izuzimaju u trenutku potpunog zdravlja. Nakon uzimanja, uzorci moraju biti obrađeni u roku od dva sata. Prvo se razrjeđuju s 1xPBSom u omjeru 1:1. Nakon toga dodajemo otopinu u epruvetu u koju smo već dodali Histopaque 1119 i Histopaque 1077 i to u omjeru 1:1:2. Nakon centrifugiranja 30 minuta pri 700xg i sobnoj temperaturi, odvojimo bijelo-mliječni sloj u kojem se nalaze mononuklearne stanice. ELISA je imunokemijska metoda pomoću koje vršimo detekciju i kvantifikaciju tvari od interesa. Jedan od reaktanata je uvijek vezan na čvrstu fazu gdje dolazi do vezanja, a tvorac signala, koji kasnije očitavamo, je enzim. Rezultati: ELISA provedena na uzorcima u ovom radu pokazala je da su razine i S100B proteina i neuron specifične enolaze podjednako povišene kod migrenaša i to za vrijeme napada migrene. Nadalje pokazuju i povišene koncentracije S100B proteina i u razdoblju bez boli. Zaključak: U ovom radu dokazana je povezanost biljega S100B i NSE s migrenom.Introduction: Migraines are one of the leading diseases of our time. The biggest problem is the diagnosis of migraines. We examine the connection between the two markers of damaged brain tissue with migraine. Objectives: Our aim is to determine the expression of these two markers and their correlation with years of peculiarities of the clinical picture of patients with migraine headache. Materials and methods: The samples of venous blood were taken from people who were separated in two groups- the examinee group and control group. Criteria for examinee group were: history of presentation of migraine headaches, age under 40 years and sampled blood in between two hours of the beginning of the attack. Exclusion criteria were: age over 40 years and other organic and pyschiatric diseases (excluding depression). Only healthy people can be in the control group. This means that they don't have migraines or any other diseases. Their samples are taken in the moment of full health. Samples have to be processed in between two hours of sampling. They are then diluated with 1xPBS. After that we add solution on top of the Histopaque 1119 and Histopaque 1077. Test tube then goes into the centrifuge. It is centrifugated 30 minutes on 700xg and room temperature. The solution is now divided in layers and the PBMCs are found in white layer. ELISA is immunochemical method by which we can identify and quantify substances of interest. One of the reactantcs is always bind to the solid phase. Enzym makes detectable signal when reaction is successful. Results: ELISA has shown us significantly higher concentrations of both markers in time of migraine attack. Also, concentration of protein S100B was significantly higher in pain free period. Conclusion: In this paper we established the connection between these markers and migraine
