2.4. Biochemical analyses
For the measurement of neurobiological markers, fasting blood samples were collected in 10 mL anticoagulant tubes between 9 and 10 a.m.. The venous blood samples were centrifuged at 3000 rpm for 15 min to be separated from serum and stored at −80°C until analyzed.
Serum samples were analyzed for TGF-1β, TNF-α, IL-1β, IL-2, IL-6, IL-10, IL-17, neopterin, BH4 and NO levels by using enzyme-linked immunosorbent assay (ELISA).
In serum samples, BH4 (Catalogue Number(CN): E-EL-0110), NO (CN: E-BC-K036), TGF-1β (CN: E-EL-H0110), IL-2 (CN: E-EL-H0099), IL-17 (CN: E-EL-H0105) levels were studied with commercial (Elabscience, Houston, Texas, USA) kits; TNF-α (CN: KAP1751), IL-1β (CN: KAP1211), IL-6 (CN: KAP1261), IL-10 (CN: KAP1321), levels were with studied commercial (Dia Source, Ottignies-Louvain-la-Neuve, Belgium) kits; neopterin (CN: RE59321) levels were studied with commercial (IBL international, Hamburg, Germany) kits.
During the analysis, ELISA washing operations were performed with an automatic washing device (BioTek ELx50 BioTek Instruments Inc. Highland Park, Winooski, VT, USA), absorbance readings were carried out in the ELISA reader (BioTekEpoch, BioTek Instruments Inc. Highland Park, Winooski, VT, USA).