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Najjar F, Rizk F, Carnac G, Nassar R, Jabak S, Sobolev AP, Bou Saada Y, El Sabban M, Hamade A.2017. Protective effect of Rhus coriaria fruit extracts against hydrogen peroxide-induced oxidative stress in muscle progenitors and zebrafish embryos. PeerJ Preprints5:e3383v1https://doi.org/10.7287/peerj.preprints.3383v1
Background and purpose Oxidative stress is involved in normal and pathological functioning of skeletal muscle. Protection of myoblasts from oxidative stress may improve muscle contraction and delay aging. Here we studied the effect of R. coriaria sumac fruit extract on human myoblasts and zebrafish embryos in conditions of hydrogen peroxide-induced oxidative stress.
Study design and Methods Crude ethanolic 70 % extract (CE) and its fractions was obtained from sumac fruits. The composition of sumac ethyl acetate EtOAc fraction was studied by 1H NMR. The viability of human myoblasts treated with CE and the EtOAc fraction was determined by trypan blue exclusion test. Oxidative stress, cell cycle and adhesion were analyzed by flow cytometry and microscopy. Gene expression was analyzed by qPCR.
Results The EtOAc fraction (IC50 2.57 μg/mL) had the highest antioxidant activity and exhibited the best protective effect against hydrogen peroxide-induced oxidative stress. It also restored cell adhesion. This effect was mediated by superoxide dismutase 2 and catalase. Pre-treatment of zebrafish embryos with low concentrations of the EtOAc fraction protected them from hydrogen peroxide-induced death in vivo. 1H NMR analysis revealed the presence of gallic acid in this fraction.
ConclusionRhus coriaria extracts inhibited or slowed down the progress of skeletal muscle atrophy by decreasing oxidative stress via superoxide dismutase 2 and catalase-dependent mechanisms.
Hatching of zebrafish embryos treated with sumac crude extract and ethyl acetate fraction at 96hpf
Hatching rates of the embryos were completed after 96 hpf of crude extract (A) and ethyl acetate fraction (B) exposure. This experiment was performed 3 times. The significance (*p ≤ 0.05, **p≤ 0.01& *** p<0.001) of embryos hatching in exposed embryos with respect to untreated embryos (CTRL).
Viability of zebrafish embryos after exposure to H2O2
Effect of H2O2 on zebrafish embryos viability after treatment with the following concentrations 10-3, 5. 10-3, 7.5 10-3, 10-2, 2. 10-2, 2.5 10-2, 2.5 10-2, 3. 10-2, 5.10-2, 7.5 10-2, 10-1. Mean values with S.D. are indicated. This experiment was performed in duplicates and repeated 3 times.