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Published on: December 11, 2020
Protection by polyphenol extract from olive stones against apoptosis produced by oxidative stress in human
Ernesto Cortés-Castell1, Carmen Veciana-Galindo, Luis Torró-Montell
1Department of Pharmacology, Pediatrics and Organic Chemistry. ernestocort@gmail.com.
Objective:
We evaluated the protective activity of an extract from a by-product such as olive stones, through its ability to inhibit H202 induced apoptosis in the SH-SY5Y human neuroblastoma cell line.
Material And Methods:
To such end, 20,000 cells/well were cultivated and differentiation with retinoic acid was initiated. Once the cells were differentiated, apoptosis was induced with and without H2O2 extract. Finally, cDNA extraction was performed, and pro-apoptotic genes Bax and anti-apoptotic genes Bcl-2 were analyzed. Quantification of the gene expression was performed using the GAPDH gene marker.
Results:
Cell viability with the extract is 97.6% (SD 5.7) with 10 mg/l and 62.8% (SD 1.2) to 50 mg/l, using 10 mg/l for the biomarker assay. The retinoic acid differentiated SH-S cell line (10 μM) shows a clear apoptosis when treated with H2O2 150 μM, with a Bax/Bcl-2 ratio of 3.75 (SD 0.80) in contrast to the differentiated control cells subjected to H2O2 and with extract, which have the same ratio of 1.02 (SD 0.01-0.03).
Conclusion:
The olive stone extract shows anti-apoptotic activity in the provoked cell death of SH-SY5Y human neuroblastoma cells in their normal state, defending them from oxidative stress which produces a significant increase in the apoptotic gene ratio in contrast to anti-apoptotic genes (Bax/Bcl-2).
Insights
Olive stone extract protects human neuroblastoma cells from oxidative stress-induced apoptosis. This extract demonstrated significant anti-apoptotic activity by modulating the Bax/Bcl-2 gene ratio.
Area of Science:
- Neuroscience
- Cell Biology
- Pharmacology
Background:
- Oxidative stress is implicated in neurodegenerative diseases.
- Neuroblastoma cell lines are utilized as models for studying neuronal function and disease.
- Olive by-products are explored for their bioactive compounds.
Purpose of the Study:
- To evaluate the protective effects of an olive stone extract against hydrogen peroxide (H2O2)-induced apoptosis.
- To investigate the extract's impact on apoptosis-related gene expression in SH-SY5Y neuroblastoma cells.
Main Methods:
- SH-SY5Y human neuroblastoma cells were differentiated using retinoic acid.
- Apoptosis was induced using H2O2, with and without the olive stone extract.
- Gene expression analysis of pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) genes was performed using quantitative real-time PCR with GAPDH as a reference gene.
Main Results:
- The olive stone extract showed dose-dependent effects on cell viability.
- Treatment with H2O2 significantly increased the Bax/Bcl-2 ratio, indicating apoptosis.
- The extract treatment significantly reduced the Bax/Bcl-2 ratio in H2O2-treated cells, suggesting an anti-apoptotic effect.
Conclusions:
- Olive stone extract possesses significant anti-apoptotic properties.
- The extract protects SH-SY5Y cells from H2O2-induced oxidative stress and apoptosis.
- This study highlights the potential therapeutic value of olive by-products in neuroprotection.
