Biochemical assessments:
Blood samples of all the subjects were taken, and aliquot samples of
serums were saved after centrifugation (20 min, 3000 rpm) at -80°C.
According to the method of Benzie & Strain in 199615, TAC was analyzed using fluorescence
recovery after photobleaching (FRAP) assay, which depends
on the capacity of serum to reduce Fe3+ to
Fe2+.
Serum MDA levels were determined by the thiobarbituric acid reactive
substances test (TBARS) spectrophotometric test, as described by Santos
in 1980 16.
PAI-1 was measured as an indicator of ED by ELIZA kit (Germany, ZellBio,
ZB-11159C-H9648). Also, VCAM-1, as another indicator of ED, was measured
by the ELIZA kit (France, Diaclone SAS, 25020).
Serum TNF-α was measured through the ELISA method according to the
manufacture’s instruction (Biovendor, Germany, Cat≠
RAF128R).