Effect of Ilex extracts and isolated compounds on peroxidase secretion of rat submandibulary glands

Free radicals are involved in diverse disorders such as tumoral, central nervous system alterations, immunological and inflammatory pathologies. Peroxidase is an oral enzyme involved in the defense of the oral cavity. Ilex species such as Ilex paraguariensis St. Hil. and the commercial product made with it “Yerba Mate” are used traditionally as antirheumatics and for the treatment of gastrointestinal diseases among others and also as a beverage with nutritional and stimulant properties. The presence of polyphenolic derivatives and flavonoids in the aqueous extract has been determined by HPLC analysis. In this study, the activity of aqueous extracts of I. paraguariensis and “Yerba Mate” on peroxidase secretion in female rat submandibular glands was investigated. The contribution to this pharmacological activity by some major hydrocynnamic acid derivatives present in the crude extracts, such as chlorogenic acid and caffeic acid and the most abundant methylxanthine, caffeine, was also evaluated. Spectrophotometrical determination of peroxidase activity showed that both extracts produced a significant increase in both secreted peroxidase and total peroxidase activity, though “Yerba Mate” showed a higher activity (EC50 “Yerba Mate”: 148 ± 10 μg/ml; EC50I. paraguariensis: 841 ± 20 μg/ml). The HPLC/DAD analysis of the crude extracts was performed and chlorogenic acid, caffeic acid and caffeine were identified and quantified. The results (expressed as W/W percentage of dried material) were as follows: I. paraguariensis: chlorogenic acid: 2.80 ± 0.30, caffeic acid: 0.023 ± 0.004, caffeine: 1.06 ± 0.06; “Yerba Mate”: chlorogenic acid: 1.98 ± 0.37; caffeic acid: 0.020 ± 0.003, caffeine: 0.70 ± 0.06. Caffeine and chlorogenic acid were proved to play an important role in the induction of peroxidase secretion induced by the extracts.