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<p>Reactive oxygen species (ROS) elicit oxidative alterations in lipids, proteins, and DNA, leading to mutations, genomic instability, and persistent inflammation, which are key factors in cancer development. Oxidative stress has a dual role in cancer, making redox modulation a key strategy for its prevention and treatment. This study investigates the antioxidant and anticancer activity of a peptide fraction (F1) isolated from the protein hydrolysate of <i>Euphorbia thymifolia</i>. The protein isolate was hydrolyzed using pepsin at 1:25 (E/S) with the highest hydrolysis rate observed at 180&#xa0;min and fractionated using a 3&#xa0;kDa ultrafiltration membrane. The ultrafiltered part was investigated employing reverse phase-high performance liquid chromatography (RP-HPLC), demonstrating that the peptide fraction (F1) displayed the most potent 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity among the three fractions. The HPLC chromatogram showed peaks at different retention times, and furthermore, the molecular weight of the peptides was found to be in the range of 500–1500 Da, with 14 peptides identified through LC-MS/MS analysis. The antioxidant properties of the peptide fraction (F1) were assessed using both the ferric reducing antioxidant power (FRAP) assay and the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay were found to be 800 ± 0.039 and 500 ± 1.23&#xa0;µg/mL, respectively. The cytotoxicity activity of the peptide fraction was found to be 80&#xa0;µg/mL. The intracellular ROS activity using 2,7’-Dichlorodihydrofluorescein diacetate (DCFDA) was found to be higher in the peptide treatment group in comparison with the group treated with H<sub>2</sub>O<sub>2</sub>, and the peptide arrested the G2/M phase of cell growth. These findings highlight the potential of the isolated peptide fraction, which could be used as a nutraceutical for treating oxidative stress-related conditions in the future. These findings underscore the potential of <i>Euphorbia thymifolia</i> peptides as a promising nutraceutical supplement for mitigating oxidative stress and promoting anticancer activity.</p> Graphical Abstract <p></p>

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Evaluation of Antioxidant and Anticancer Properties of Euphorbia thymifolia Protein Hydrolysates Using in Vitro Studies

  • 期刊论文

  • 发表日期：

    2025年8月19日

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  • S. Jagadeeshwari,
  • S. Rupachandra

摘要

Reactive oxygen species (ROS) elicit oxidative alterations in lipids, proteins, and DNA, leading to mutations, genomic instability, and persistent inflammation, which are key factors in cancer development. Oxidative stress has a dual role in cancer, making redox modulation a key strategy for its prevention and treatment. This study investigates the antioxidant and anticancer activity of a peptide fraction (F1) isolated from the protein hydrolysate of Euphorbia thymifolia. The protein isolate was hydrolyzed using pepsin at 1:25 (E/S) with the highest hydrolysis rate observed at 180 min and fractionated using a 3 kDa ultrafiltration membrane. The ultrafiltered part was investigated employing reverse phase-high performance liquid chromatography (RP-HPLC), demonstrating that the peptide fraction (F1) displayed the most potent 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity among the three fractions. The HPLC chromatogram showed peaks at different retention times, and furthermore, the molecular weight of the peptides was found to be in the range of 500–1500 Da, with 14 peptides identified through LC-MS/MS analysis. The antioxidant properties of the peptide fraction (F1) were assessed using both the ferric reducing antioxidant power (FRAP) assay and the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay were found to be 800 ± 0.039 and 500 ± 1.23 µg/mL, respectively. The cytotoxicity activity of the peptide fraction was found to be 80 µg/mL. The intracellular ROS activity using 2,7’-Dichlorodihydrofluorescein diacetate (DCFDA) was found to be higher in the peptide treatment group in comparison with the group treated with H2O2, and the peptide arrested the G2/M phase of cell growth. These findings highlight the potential of the isolated peptide fraction, which could be used as a nutraceutical for treating oxidative stress-related conditions in the future. These findings underscore the potential of Euphorbia thymifolia peptides as a promising nutraceutical supplement for mitigating oxidative stress and promoting anticancer activity.

Graphical Abstract