Betulinic acid mitigates oxidative stress-mediated apoptosis and enhances longevity in the yeast Saccharomyces cerevisiae model

Abstract

Betulinic acid (BA), a pentacyclic triterpenoid found in certain plant species, has been reported to have several health benefits including antioxidant and anti-apoptotic properties. However, the mechanism by which BA confers these properties is currently unknown. Saccharomyces cerevisiae, a budding yeast with a short life cycle and conserved cellular mechanism with high homology to humans, was used as a model for determining the role of BA in aging and programmed cell death (PCD). Treatment with hydrogen peroxide (H₂O₂) exhibited significantly increased (30–35%) survivability of antioxidant (sod1Δ, sod2Δ, cta1Δ, ctt1Δ, and tsa1Δ) and anti-apoptotic (pep4Δ and fis1Δ) mutant strains when cells were pretreated with BA (30 µM) as demonstrated in spot and CFU (Colony forming units) assays. Measurement of intracellular oxidation level using the ROS-specific dye H₂DCF-DA showed that all tested BA-pretreated mutants exhibited decreased ROS than the control when exposed to H₂O₂. Similarly, when mutant strains were pretreated with BA and then exposed to H₂O₂, there was reduced lipid peroxidation as revealed by the reduced malondialdehyde content. Furthermore, BA-pretreated mutant cells showed significantly lower apoptotic activity by decreasing DNA/nuclear fragmentation and chromatin condensation under H₂O₂-induced stress as determined by DAPI and acridine orange/ethidium bromide staining. In addition, BA treatment also extended the life span of antioxidant and anti-apoptotic mutants by ∼10–25% by scavenging ROS and preventing apoptotic cell death. Our overall results suggest that BA extends the chronological life span of mutant strains lacking antioxidant and anti-apoptotic genes by lowering the impact of oxidative stress, ROS levels, and apoptotic activity. These properties of BA could be further explored for its use as a valuable nutraceutical.

Keywords:

• Betulinic acid
• aging
• apoptosis
• ROS
• Saccharomyces cerevisiae

Acknowledgement

This work was supported by DBT-BIRAC supported project (BIRAC/BT/SB0066/02/19) to DAN. The authors express their sincere gratitude to the Director, CSIR-CIMAP for his support throughout the study. The authors are also thankful to Dr. Ravi Manjithaya for providing the mutant strains tsa1Δ, cta1Δ, pep4Δ, and ctt1Δ, and Dr. P. Vijayraj for providing yeast mutants sod1Δ, sod2Δ, ctt1Δ, and fis1Δ.

Disclosure statement

Authors declare no conflict of interest. Institutional communication number for this article is CIMAP/PUB/2022/73.

Additional information

Funding

This work was supported by Biotechnology Industry Research Assistance Council [Grant No. BIRAC/BT/SB0066/02/19].