The present study aims to investigate the effects of a standardized raw extract of C. asiatica (RECA) on hydrogen peroxide (H2O2)-induced oxidative stress and apoptotic death in neural-like cells derived from mouse embryonic stem (ES) cell line.
A transgenic mouse ES cell (46C) was differentiated into neural-like cells using 4-/4+ protocol with addition of all-trans retinoic acid. These cells were then exposed to H2O2 for 24 h. The effects of RECA on H2O2-induced neural-like cells were assessed through cell viability, apoptosis, and reactive oxygen species (ROS) assays, as well as neurite length measurement. The gene expression levels of neuronal-specific and antioxidant markers were assessed by RT-qPCR analysis.
Pre-treatment with H2O2 for 24 hours, in a dose-dependent manner, damaged neural-like cells as marked by a decrease in cell viability, substantial increase in intracellular ROS accumulation, and increase in apoptotic rate compared to untreated cells. These cells were used to treat with RECA. Treatment with RECA for 48 h remarkably restored cell survival and promoted neurite outgrowth in the H2O2- damaged neurons by increasing cell viability and decreasing ROS activity. RT-qPCR analysis revealed that RECA upregulated the level of antioxidant genes such as thioredoxin-1 (Trx-1) and heme oxygenase-1 (HO-1) of treated cells, as well as the expression level of neuronal-specific markers such as Tuj1 and MAP2 genes, suggesting their contribution in neuritogenic effect.
Our findings indicate that RECA promotes neuroregenerative effects and exhibits antioxidant properties, suggesting a valuable synergistic activity of its phytochemical constituents, thus, making the extract a promising candidate in preventing or treating oxidative stress-associated Alzheimer\'s disease.
本研究旨在研究标准化的积雪草原提取物(RECA)对过氧化氢(H2O2)诱导的氧化应激和源自小鼠胚胎干(ES)细胞系的神经样细胞凋亡的影响。
使用加入全反式视黄酸的4-/4+方案将转基因小鼠ES细胞(46C)分化成神经样细胞。然后将这些细胞暴露于H2O224小时。通过细胞活力评估RECA对H2O2诱导的神经样细胞的影响。凋亡,和活性氧(ROS)测定,以及神经突长度测量。通过RT-qPCR分析评估神经元特异性和抗氧化剂标志物的基因表达水平。
用H2O2预处理24小时,以剂量依赖的方式,受损的神经样细胞,以细胞活力下降为标志,细胞内ROS积累的大量增加,与未处理的细胞相比,凋亡率增加。这些细胞用于用RECA处理。用RECA处理48小时可显着恢复细胞存活,并通过增加细胞活力和降低ROS活性来促进H2O2损伤神经元的神经突生长。RT-qPCR分析显示,RECA上调了处理细胞的抗氧化基因如硫氧还蛋白-1(Trx-1)和血红素加氧酶-1(HO-1)的水平,以及神经元特异性标志物如Tuj1和MAP2基因的表达水平,表明它们在神经源性效应中的作用。
我们的研究结果表明,RECA促进神经再生作用并表现出抗氧化特性,表明其植物化学成分具有有价值的协同活性,因此,使提取物成为预防或治疗氧化应激相关的阿尔茨海默病的有希望的候选物。