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目的:探讨淫羊藿总黄酮(Total epimedium flavonoids,TEF)对喹啉酸(Quinolinic acid,QA)诱导的人神经母细胞瘤细胞(SH-SY5Y)毒性的保护作用及其可能机制。方法:噻唑蓝(MTT)比色法测定QA的毒性作用;不同浓度淫羊藿总黄酮预处理SH-SY5Y细胞,再用QA共孵育,用MTT比色法测定细胞存活率;用Hochest33258染色观察细胞凋亡;用比色法测定超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量。用流式细胞技术(Flow cytometry,FCM)检测细胞线粒体膜电位(Mitochondrial membrane potential,△Ψm)和细胞内Ca2+浓度([Ca2+]i)的变化。结果:淫羊藿总黄酮能提高细胞的存活率;使呈现细胞核聚集浓缩、致密浓染的凋亡细胞显著减少;提高SOD和GSH-Px活性,降低MDA含量;升高线粒体膜电位(△Ψm);降低细胞内钙离子浓度([Ca2+]i)。结论:淫羊藿总黄酮对QA诱导的SH-SY5Y细胞毒性有显著的保护作用。
Objective: To investigate the protective effect of total epimedium flavonoids (TEF) on the toxicity of Quinolinic acid (QA) induced human neuroblastoma cells (SH-SY5Y) and its possible mechanism. Methods: Toxicity of QA was determined by MTT colorimetric assay. SH-SY5Y cells were pretreated with different concentrations of Epimedium sagittatum and co-incubated with QA. Cell viability was determined by MTT colorimetric assay. Hochest33258 staining Cell apoptosis was measured. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were determined by colorimetry. The changes of mitochondrial membrane potential (△ Ψm) and intracellular Ca2 + concentration ([Ca2 +] i) were detected by flow cytometry (FCM). Results: Epimedium sagittatum flavonoids could increase the survival rate of cells. The number of apoptotic cells that aggregated and densely condensed in the nucleus was significantly decreased. The activities of SOD and GSH-Px were increased, the content of MDA was decreased, the mitochondrial membrane potential (△ ψm ); Reduce the intracellular calcium concentration ([Ca2 +] i). Conclusion: Epimedium Flavonoids have a significant protective effect on QA-induced SH-SY5Y cytotoxicity.