Baskaran Rathinasamy, Kalaiselvi Palanisamy, Huang Chih-Yang, Padma Viswanadha Vijaya
DRDO BU Center for Life Science, Bharathiar University, Coimbatore, Tamil Nadu, India.
Translational Research Laboratory, Department of Biotechnology, School of Biotechnology and Genetic Engineering, Bharathiar University, Coimbatore, Tamil Nadu, India.
Integr Med Res. 2015 Dec;4(4):231-241. doi: 10.1016/j.imr.2015.09.002. Epub 2015 Sep 30.
Neferine, a bisbenzylisoquinoline alkaloid, isolated from has a wide range of biological activities. Cobalt chloride (CoCl) was known to mimic hypoxic condition. In the present study, we assessed the cytoprotective effect of neferine against CoCl-induced oxidative stress in muscle cells.
Rhabdomyosarcoma cells were exposed to different concentrations of CoCl, and the IC value was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Lactate dehydrogenase and NO assays were performed in order to determine the cytotoxic effect of CoCl. Reactive oxygen species generation and cellular antioxidant status were determined for evaluating oxidative stress. For analyzing the effect of neferine on CoCl-induced apoptosis, propidium iodide staining was performed.
The results of the present study indicate that CoCl induces cell death in a dose-dependent manner. Neferine pretreatment at 700 nM concentration offers better cytoprotection in the cells exposed to CoCl. Lactate dehydrogenase and NO release in the culture medium were restored after neferine pretreatment. CoCl triggers time-dependent reactive oxygen species generation in muscle cells. Further, results of propidium iodide staining, mitochondrial membrane potential, and intracellular calcium accumulation confirm that neferine offers protection against CoCl-induced hypoxic injury. Depleted activities of antioxidants such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase due to CoCl exposure were also reinstated in the group that received neferine pretreatment.
Our study suggests that neferine from offers protection to muscle cells by counteracting the oxidative stress induced by CoCl.
从[具体来源未给出]中分离出的双苄基异喹啉生物碱甲基莲心碱具有广泛的生物活性。已知氯化钴（CoCl）可模拟缺氧状态。在本研究中，我们评估了甲基莲心碱对CoCl诱导的肌肉细胞氧化应激的细胞保护作用。
将横纹肌肉瘤细胞暴露于不同浓度的CoCl中，使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法测定IC值。进行乳酸脱氢酶和NO测定以确定CoCl的细胞毒性作用。测定活性氧生成和细胞抗氧化状态以评估氧化应激。为分析甲基莲心碱对CoCl诱导的细胞凋亡的影响，进行碘化丙啶染色。
本研究结果表明CoCl以剂量依赖性方式诱导细胞死亡。700 nM浓度的甲基莲心碱预处理能为暴露于CoCl的细胞提供更好的细胞保护。甲基莲心碱预处理后，培养基中的乳酸脱氢酶和NO释放得以恢复。CoCl在肌肉细胞中引发时间依赖性的活性氧生成。此外，碘化丙啶染色、线粒体膜电位和细胞内钙积累的结果证实甲基莲心碱可保护细胞免受CoCl诱导的缺氧损伤。在接受甲基莲心碱预处理的组中，由于CoCl暴露而耗尽的抗氧化剂如超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和谷胱甘肽S-转移酶的活性也得以恢复。
我们的研究表明，[具体来源未给出]中的甲基莲心碱通过抵消CoCl诱导的氧化应激为肌肉细胞提供保护。
