Antioxidant effects of lebanese Crocus sativus L. and its main components, crocin and safranal, on human skeletal muscle cells

Abstract

Introduction- Crocus sativus L. (C. sativus L.) has gained interest as a potential source of pharmacologically bioactive compounds and is believed to possess antioxidant properties. This study used human myoblast cells to evaluate the protective effects of C. sativus L. extracts, (crocin, and safranal) on hydrogen peroxide (H2O2)-induced oxidative stress using human myoblast cells. Methods- HPLC was used to detect the presence of the active compounds safranal/crocin in the Lebanese C. sativus L. extracts. Their cellular antioxidant property was examined by trypan blue, cell adhesion, immunostaining and cell cycle assays. Transcriptional expression and activities of SOD1/2, GPX1, and catalase were also determined. The ability of C. sativus L. and its main components to modulate myogenic regulatory factors (MyoD, Myf5) was evaluated using RT-PCR assay. Results- The water/methanol (50:50, v/v) extract of C. sativus L. stigmas were found to contain safranal and crocin, with a high concentration of trans-crocin 3/4 in particular. In vitro, pretreatment (24 h) with safranal exhibited the lowest antioxidant effect whereas pretreatment with C. sativus L. extracts (0.3 µg/ml) and more notably with crocin (0.3 µM) attenuated the toxic impact of H2O2 (50 µM, 24 h) and also restored the capacity of adhesion among LHCN-M2 cells. This protective effect was associated with both a reduction of intracellular reactive oxygen species generation and an up-regulation of antioxidant enzyme activities. Furthermore, crocin pretreatment significantly improved the transcriptional expression of myogenic regulatory genes. Conclusion- C. sativus L., particularly its main component crocin, was shown to have therapeutic potential against oxidative stress-associated skeletal muscle diseases.