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Ex vivo protective effect of rosmarinic acid and Mentha×villosa water extract on tissue injury induced by ischaemia/reperfusion
Ischaemia/reperfusion (I/R) is known to induce tissue injury associated with increased production of reactive oxygen species (ROS). Thus compounds with antiradical activity may prove beneficial as they help to reduce or even to prevent this damage. The aim of the study was to test the effect of the water extract of leaves of Mentha×villosa Huds (EMV, 1.3mg/10ml)) and its main phenolic active compound – rosmarinic acid (RA, 5×10–4 mol/l) on changes in ROS production induced by I/R in intestinal and vascular tissues. In anaesthetised rats, ischaemia was induced in vivo by clamping the onset of the superior mesenteric artery (SMA) for 60min followed by 30min reperfusion1. After sacrifying animals, the ileum and SMA were removed. Tissue segments were placed into the testing solutions with or without EMV and RA and incubated for 30min (SMA) or 60min (ileum). Tissues of sham-operated animals served as a control. Production of ROS by segments of the ileum and SMA was determined using luminol enhanced chemiluminiscence (CL). After I/R, CL in SMA increased from control values of 7.9±1.66 to 15.21±2.49 mV/mg w.w., in intestinal segments from 33.74±6.47 to 58.53±11.73 mV/100mg w.w. Both, EMV and RA reduced intestinal and vascular CL to control values. Moreover in in vitro experiments, the antioxidant activity of EMV and RA was tested using DPPH test. SC50% values were 7.26±0.09µg/ml and 1.72±0.13µg/ml, respectively. In ABTS test SC50% were 15.07±0.47µg/ml and 1.93±0.03µg/ml, respectively. The results confirmed the attenuation of increased ROS production induced by I/R in SMA and ileum by rosmarinic acid and Mentha×villosa extract, and thus indicate their probable protective effect on tissues.
Acknowledgements: VEGA grant MS SR 2/0050/09
References: 1. Sotníková, R. et al. (2005) Biologia (17) 60: p 145–147.