Effect of Hypogymnia physodes Extracts and their Depsidones on Micronucleus Distribution in Human Lymphocytes

Abstract

Three lichen depsidones, physodalic acid (1), physodic acid (2), and 3-hydroxy physodic acid (3), were isolated from Hypogymnia physodes diethyl ether extract using column chromatography, and their structures determined by comparing their UV, 1 H and 13 C NMR spectroscopic and MS data with those given in the literature, as well as with data computed by CHEM draw ultra 11 software. The contents of 1, 2 and 3 were determined in the methanol (ME), acetone (AE), and diethyl ether (EE) extracts using reversed-phase high performance liquid chromatography with photodiode array detection. The extracts, isolated depsidones 1-3 and EE fraction F23 (consisting of 90% 2 and 3, in the ratio 5.5: 1) were evaluated for their in vitro effects on chromosome aberrations in peripheral human lymphocytes using the cytochalasin-B blocked micronucleus (CBMN) assay in doses of 1 µg/mL and 2 µg/mL of final culture solution. The frequency of MN was scored in binucleated cells, and nuclear proliferation index was calculated. It was found that 1, 2, 3, F23, and EE at 1.0 µg/mL exerted a beneficial effect on lymphocyte cells giving a significant decrease of the frequency of MN in comparison with the positive control Amifostin WR-2721.Among the tested depsidones at a concentration of 1 µg/mL, 3 exhibited the most prominent effect decreasing the frequency of MN by 30.3%, followed by 2 (28.2%) and 1 (22.0%). The extracts were less effective than the isolated depsidones.

Ramalina capitata (Ach.) Nyl. acetone extract: HPLC analysis, genotoxicity, cholinesterase, antioxidant and antibacterial activity

“…Cytokinesis - block micronucleus assay was performed as previously described (Fench and Morley, 1993[ 7 ]; Stojanović et al, 2013[ 25 ]). The cell culture lymphocytes were treated with 1.0, 2.0 and 3.0 μg mL -1 of the examined acetone extracts.…”

Lichen Depsidones with Biological Interest

“…This compound also diminished the proliferation of thymocytes inducing cytotoxicity via ROS production. Physodalic acid reduced the frequency of micronucleus (30.3 %) on lymphocytes [71,104].…”

“…Moreover, physodic acid reduced thymocyte proliferation-induced cytotoxicity via oxidative stress mainly through ROS production [71]. On lymphocytes, this depsidone significantly decreased micronucleus frequency (28.2 %) compared to the positive control [104]. Furthermore, this compound proved to significantly reduce human cancer HeLa cell viability (IC 50 [24 h] value of 171 μg/mL and IC 50 [72 h] value of 63 μg/ mL) [105].…”

Clastogenic Effect of Atranorin, Evernic acid, and Usnic Acid on Human Lymphocytes

“…Our previous paper [3] showed that lichen depsidones, physodalic acid, physodic acid, and 3-hydroxy physodic acid, also decreased the frequency of MN at a concentrations of 1.0 and 2.0 µg/mL more than amifostine at a concentration of 1 µg/mL. However, their effects were smaller than the effect of usnic acid.…”

“…Continuing our investigation on the biological activities of the lichen substances [2,3], herein we report the clastogenic effect (micronucleus distribution) and antiproliferative effect (CBPI) in peripheral blood lymphocytes of the atranorin (isolated from Hypogymnia physodes), evernic acid (isolated from Evernia prunastri), and (+)-usnic acid (isolated from Usnea barbata). The structures of examined compounds 1, 2, and 3, as well as structures of positive control (amifostine) and negative control (mitomycin C, MMC) are given on Figure 1.…”