Saha Sarmistha, Goswami Gagan, Pandrangi Anupama
Department of Zoology, University School of Sciences, Gujarat University, Ahmedabad 380009, India.
Department of Zoology, University School of Sciences, Gujarat University, Ahmedabad 380009, India.
Chem Biol Interact. 2014 Dec 5;224:51-7. doi: 10.1016/j.cbi.2014.09.016. Epub 2014 Oct 16.
Calcium oxalate monohydrate (COM) has been shown to be the most frequent constituent of kidney stones. The interactions of cells with COM crystals produce a variety of physiological and pathological changes including the development of oxidative stress, cellular injury and apoptosis. On the other hand, diosgenin, a steroidal sapogenin, is well known for its antioxidant activity. Therefore, the aim of this study was to evaluate whether diosgenin protects MDCK renal epithelial cells from COM-induced apoptotic death. Diosgenin was isolated from fruits of Solanum xanthocarpum by silica gel column chromatography. It was obtained in high yields (1.23%) and the purity was ascertained by HPTLC analysis. Characterization of diosgenin was done by mp, UV-visible spectrophotometry, elemental analysis, FT-IR, (1)H NMR and (13)C NMR analysis. Cells were co-incubated with COM (80μg/cm(2)) and diosgenin (2.5, 5, 7.5 and 10μg/mL) for 24h. It was found that diosgenin attenuated the apoptotic death induced by COM as measured in terms of cell viability, caspase -9/3 activities and DNA fragmentation percent. The inhibitory role of diosgenin on caspase -9/3 activities was also analyzed using molecular docking experiments, which showed interactions to their active sites by H-bonds. Diosgenin also attenuated the increase in lipid peroxidation and glutathione depletion induced by COM crystals. In conclusion, the preventive effect of diosgenin is associated to the inhibition of oxidative stress and caspases.
一水合草酸钙（COM）已被证明是肾结石中最常见的成分。细胞与COM晶体的相互作用会产生多种生理和病理变化，包括氧化应激的发展、细胞损伤和凋亡。另一方面，薯蓣皂苷元是一种甾体皂苷元，以其抗氧化活性而闻名。因此，本研究的目的是评估薯蓣皂苷元是否能保护MDCK肾上皮细胞免受COM诱导的凋亡死亡。通过硅胶柱色谱法从黄果茄果实中分离出薯蓣皂苷元。其产率较高（1.23%），并通过高效薄层色谱分析确定了纯度。通过熔点、紫外-可见分光光度法、元素分析、傅里叶变换红外光谱、核磁共振氢谱和核磁共振碳谱分析对薯蓣皂苷元进行了表征。将细胞与COM（80μg/cm²）和薯蓣皂苷元（2.5、5、7.5和10μg/mL）共同孵育24小时。结果发现，从细胞活力、半胱天冬酶-9/3活性和DNA片段化百分比方面衡量，薯蓣皂苷元减轻了COM诱导的凋亡死亡。还使用分子对接实验分析了薯蓣皂苷元对半胱天冬酶-9/3活性的抑制作用，结果表明它通过氢键与它们的活性位点相互作用。薯蓣皂苷元还减轻了COM晶体诱导的脂质过氧化增加和谷胱甘肽消耗。总之，薯蓣皂苷元的预防作用与抑制氧化应激和半胱天冬酶有关。
