Effect of α-Tocopherol During In Vitro Ozonation of Methyl Linoleate: Its Implication in Ozone Therapy

Abstract

ABSTRACT Systemic ozone therapy is widely used as an oxidant therapy to treat many conditions and diseases. It is known that ozone therapy acts through a transient oxidative stress produced by lipid ozonation products. The convenience of supporting patients with antioxidants during systemic ozone therapy applications is now under discussion. We studied the reaction of linoleate (one of the main constituents of cellular membranes and plasma phospholipids) with ozone in presence or absence of α-tocopherol, in order to explore whether the combination of ozone and antioxidant has some effect on fatty acid ozonation products. Proton nuclear magnetic resonance spectroscopy (1H NMR) was used for following the reaction between 4.8mol mmol of methyl linoleate and 2.08 or 4.32mmol of ozone, with addition of different amounts of α-tocopherol (0.10, 0.18, and 0.26μmol). Ozonide (δ=5.15ppm) and aldehydes (δ=9.63ppm and δ=9.74ppm) intensities from 1H NMR signals markedly decreased with α-tocopherol addition. When αtocopherol is absent, the intensities from olefinic proton signals diminished with ozone concentration increment; however, with αtocopherol in the mixture a smaller decrement was achieved. No detectable signals were found with the ozonation of α-tocopherol without methyl linoleate in the reaction mixture. These results suggest that α-tocopherol reacts with those products released from the reaction of ozone with methyl linoleate. This fact points out that antioxidant supplementation during systemic ozone therapy (major and minor autohemotherapy, rectal insufflation, and so forth) can be detrimental toward achieving the needed transient oxidative stress responsible for biological activities.