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).