Cortés-Castell Ernesto, Veciana-Galindo Carmen, Torró-Montell Luis, Palazón-Bru Antonio, Sirvent-Segura Elia, Gil-Guillén Vicente, Rizo-Baeza Mercedes
Department of Pharmacology, Pediatrics and Organic Chemistry.
Nutr Hosp. 2016 Feb 16;33(1):118-22. doi: 10.20960/nh.39.
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.
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.
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).
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).
我们通过评估橄榄核等副产品提取物抑制H2O2诱导人神经母细胞瘤SH-SY5Y细胞系凋亡的能力，来评价其保护活性。
为此，每孔接种20000个细胞并开始用视黄酸诱导分化。细胞分化后，分别在有和无H2O2提取物的情况下诱导凋亡。最后，进行cDNA提取，并分析促凋亡基因Bax和抗凋亡基因Bcl-2。使用GAPDH基因标记物对基因表达进行定量分析。
提取物浓度为10mg/l时细胞活力为97.6%（标准差5.7），50mg/l时为62.8%（标准差1.2），使用10mg/l进行生物标志物检测。视黄酸分化的SH-S细胞系（10μM）在150μM H2O2处理时显示明显凋亡，Bax/Bcl-2比值为3.75（标准差0.80），而接受H2O2和提取物处理的分化对照细胞的该比值为1.02（标准差0.01 - 0.03）。
橄榄核提取物在正常状态下对SH-SY5Y人神经母细胞瘤细胞的诱导性细胞死亡具有抗凋亡活性，使其免受氧化应激影响，与抗凋亡基因（Bax/Bcl-2）相比，氧化应激会导致凋亡基因比值显著增加。
