Environmental genome project: focusing on differences to understand the whole.

Corresponding Author Tel: (+39) 0577 234226 Fax: (+39) 0577 234219 Email: [email protected] Backg ro und: Em pirical observations have shown th at ozonated autohem otherapy m arkedly im proves the sym ptom s of chron ic lim b ischem ia (m uscular pain at rest, interm itten t claudication , e tc ) in ath erosclerotic patien ts , but m echan ism s of action rem ain unclear. Aims : Hum an endothelia l ce lls (HUVECs ) are known to release nitrogen m onox ide (NO) and we inve stigated th e biological effe cts of hum an ozonated serum on HUVECs in culture. Methods : We assess ed the re levance of pe rox idation, the release of NO as nitr ite and of th ree clas s ical cytokin es . Res ults : Th e treatm ent of HUVECs w ith ozonated se rum yie lds a dos e dependen t increas e of th iobarbituric acid reactive subs tances (TBARS) and of hydrogen perox ide (H2O2 ) and a decrease of protein th iol groups (PTG). Concom itan tly, in com par ison to eithe r the con trol or the ox ygenated sam ple, the re is a s ign ific an t and steady in crease of n itr ic ox ide (NO) production ; th is is m arkedly enhanced by the addition of L-argin in e (20 m M) and inh ibited in th e pre sence of the NO inh ibitor, L-NAME (20 m M). The m ain m edia tor of ozone action is H2O2 as it has be en show n eithe r afte r its direct m easurem ent or by the additio n of 20, 40 and 100 m M. Moreover, during 24 hours incubation w e have in vestiga ted th e production of endoth elin 1 (ET-1 ), E-selectin and In te rleukin 8 (IL-8 ) and it appears th at ozonation enhances IL-8, in h ibits E-selectin and hardly m odifie s ET-1 production . Conc lus ions : It appears th at r ein fus ion of ozonated blood, by enhancin g re lease of NO, m ay in duce vasodilation in ischem ic areas and reduce hypox ia.