Additive toxicity of limonene and 50% oxygen and the role of glutathione in detoxification in human lung cells

Limonene has many commercial applications and has been introduced as an environmentally acceptable solvent replacing halogenated hydrocarbons. Occupational exposure to limonene presumably occurs simultaneously with other chemicals including oxidative agents and may exert a heavy strain on cellular detoxifying capacity resulting in synergistic effects. The present study used oxygen as an example of an ubiquitous oxidative and radical forming agent and investigated the combination effects with limonene on human lung cells. Mechanistic information was gained by comparing the toxicity of limonene with a major oxidation product, limonene 1,2-epoxide, and by the involvement of glutathione in cellular detoxification. At cell culture conditions most similar to the in vivo situation oxygen did not increase the toxicity of limonene beyond an additive effect. The results further indicated that limonene 1,2-epoxide was not the active compound in limonene toxicity. Experimental evidence suggests that detoxification of limonene in human lung cells primarily occurs by mechanisms not involving the glutathione system and point to possible long-term effects of limonene exposure. The present knowledge indicates clearly that the mechanism of action of limonene on biological systems and particularly in combination with oxidative compounds still remains to be elucidated. In light of the frequent exposure of humans to such combinations further investigations into this issue are highly recommended.