Bioactive polyphenols separated from hemp seed shells ameliorate H2 O2 -induced oxidative stress injury in human umbilical vein endothelial cells

Abstract

In the present study, we investigated the protection of hemp seed polyphenols on human umbilical vein endothelial cells (HUVEC) from H₂ O₂ -mediated oxidative stress injury. Fractions with different polarities were obtained by separating the hemp seed extract using HPD300 macroporous resin-packed column. The fraction, desorbed by 50% ethanol, is rich in polyphenol (789.51 ± 21.92 mg GAE/g) and has the highest antioxidant activity in vitro. HPLC-QTOF-MS/MS identified the main polyphenol components in hemp seed shells: 4 hydroxycinnamic acid amides and 15 lignanamides. The protective effects of hemp seed polyphenol against oxidative-stress injury in HUVEC cells were evaluated by cell viability, intracellular antioxidant parameters, and cell apoptosis assay. After HUVEC cells were precultured with 50 µg/ml hemp seed polyphenols, the cell viability increased significantly from 53.07 ± 2.46% (model group) to 80.65 ± 1.32% (p < 0.01). In addition, the pretreatment of HUVEC cells with polyphenol could substantially increase their intracellular superoxide dismutase activity and reduce their intracellular reactive oxygen species level, malondialdehyde content, and lactate dehydrogenase leakage index. These findings demonstrate the defensive potential of hemp seed polyphenol in reducing the incidence of cardiovascular disease. PRACTICAL APPLICATION: Hemp seed shell waste is produced while producing hemp seed kernel and has abundant phenolic compounds. This research showed that hemp seed polyphenol has potent antioxidant activity in vitro and protects HUVEC cells against H₂ O₂ -induced oxidative stress injury, suggesting that hemp seed polyphenol has the defensive potential to reduce the incidence of cardiovascular disease. These results indicated that polyphenol separated from hemp seed shells is valuable for further research and development, which will improve the utilization rate of hemp seed.

Keywords: antioxidant; hemp seed; human umbilical vein endothelial cells; oxidative stress; polyphenol.