Screening of new antioxidant molecules using flow cytometry

Area of Science:

• Cell Biology
• Biochemistry
• Pharmacology

Background:

• Oxidative stress impacts cellular health.
• Evaluating antioxidant properties of novel compounds is crucial.
• Existing methods may have limitations in assessing cellular antioxidant activity.

Purpose of the Study:

• To establish a flow cytometry technique for evaluating the antioxidant properties of molecules on living cells.
• To synthesize and screen novel ellipticine analogues for their antioxidative potential.
• To compare the efficacy of these analogues against established antioxidants like Vitamin E and Ebselen.

Main Methods:

• Utilized a murine-murine hybridoma (Mark 3) cell line for cell-based assays.
• Employed dihydrorhodamine (DHR-123) to measure intracellular superoxide anions.
• Used dichlorofluorescein diacetate (DCFH-DA) to quantify intracellular peroxides.
• Induced oxidative stress using hydrogen peroxide (H2O2) or the xanthine (X)/xanthine oxidase (XO) system.
• Analyzed approximately 35,000 cells per experiment for robust statistical analysis.

Main Results:

• The flow cytometry technique effectively evaluated oxidative stress in cells.
• Twenty-one novel ellipticine analogues were synthesized and tested.
• Selenated molecule 18 demonstrated 10-fold higher activity than Ebselen.
• Molecule 18's activity was significantly lower than Vitamin E.
• Eight compounds exhibited glutathione peroxidase-like activity.

Conclusions:

• The developed flow cytometry method provides a reliable platform for assessing cellular antioxidant capacity.
• Novel ellipticine analogues possess significant antioxidant properties.
• Selenated molecule 18 represents a promising candidate for further investigation due to its potent antioxidant effects.