Antioxidant activity of fractions isolated from hemolymph of garden snail Helix lucorum

Article published in the Journal of Pharmacy & Pharmacognosy Research 9(2): 143-152, 2021.

Original Article

Albena Alexandrova^1,3*, Lubomir Petrov^1,3, Lyudmila Velkova², Aleksander Dolashki², Elina Tsvetanova³, Almira Georgieva³, Pavlina Dolashka²

¹Department of Physiology and Biochemistry, National Sports Academy, 21, Acad. Stefan Mladenov, str., 1700 Sofia, Bulgaria.

²Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 9, Georgi Bonchev str., 1113 Sofia, Bulgaria.

³Laboratory of Free Radical Processes, Institute of Neurobiology, Bulgarian Academy of Sciences, 23, Georgi Bonchev str., 1113 Sofia, Bulgaria.

*E-mail: a_alexandrova_bas@yahoo.com

Abstract

Context: The snail hemolymph is a multi-component mixture comprising various substances with pharmacological activity.

Aims: To evaluate the antioxidant activity of hemocyanin and fractions isolated of hemolymph from garden snail Helix lucorum.

Methods: The antioxidant activity was tested in chemical systems generating superoxide radicals (O₂^•–) and hydroxyl radicals (^•OH) and was presented as percent inhibition. The fractions’ composition was analyzed by MALDI-TOF/TOF mass spectrometry.

Results: At concentration of 90 µg/mL the hemolymph fraction with compounds with molecular weight (MW) <100kDa demonstrated the strongest O₂^•–– scavenger effect (82%), followed by the hemocyanin (58%), and hemolymph fraction with compounds with MW<1kDa (15%). The most powerful ^•OH scavenger effect at concentration of 90 µg/mL showed hemocyanin (43%) followed by MW<100kDa fraction (26%). No ^•OH scavenger effect was observed by the MW<1kDa fraction. The highest chelation properties (55%) were demonstrated by the MW<100kDa fraction at concentration of 130 µg/mL; both the hemocyanin and MW<1kDa hemolymph fraction showed 33% inhibitory effect at the same concentration. The analysis of the composition of the tested fractions revealed high content of hydrophobic surfaces in MW<1kDa fraction and presence of antioxidant enzymes-catalase, superoxide dismutase and glutathione peroxidase in MW<100kDa fraction that contribute to the overall antioxidant effect of H. lucorum hemolymph.

Conclusions: The complex antioxidant effect of hemolymph from H. lucorum is thought to be related to the presence of low molecular weight peptides in the MW<1 kDa fraction including glutathione and mainly to the antioxidant enzymes in the MW<100 kDa fraction.