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机构地区:[1]河南科技大学医学院药理教研室,河南洛阳471003
出 处:《中国中药杂志》2013年第9期1318-1322,共5页China Journal of Chinese Materia Medica
基 金:河南省医学科技攻关计划项目(201003079)
摘 要:目的:研究芍药苷对Aβ25-35诱导PC12细胞损伤的保护作用及其机制。方法:Aβ25-35造成PC12细胞损伤,检测细胞的存活率(MTT法)、乳酸脱氢酶(LDH)漏出率以及细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)水平的变化。采用流式细胞仪测定细胞内活性氧(ROS)水平和线粒体膜电位的变化。Western blot法检测细胞内HO-1,Cyt C和cleaved Caspase-3蛋白的表达水平。结果:芍药苷组(5,10,20μmol.L-1)能不同程度地对抗Aβ25-35对PC12细胞的损伤,增加细胞存活率,减少LDH漏出率和MDA含量,增加抗氧化酶SOD,GSH-Px和HO-1的活性,减少细胞内ROS形成,提高线粒体膜电位,减少Cyt C释放和cleaved Caspase-3蛋白的表达。结论:芍药苷对Aβ25-35致PC12细胞损伤具有一定的抑制作用,其作用机制可能与抑制Aβ25-35诱导的氧化应激以及减轻线粒体损伤有关。Objective: To investigate the neuroprotective effects and mechanism of paeoniflorin on Aβ25-35-induced oxidative stress in PC12 cells. Method: The proliferation of induced PCl2 cells were investigated by the MTTmethod. The leakage rate of lac- tate dehydrogenase (LDH), the intracellular content of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione perox- idase (GSH-Px) were also measured. The changes of the intraceUular reactive oxygen species (ROS) level and the mitochondrial membrane potential (MMP) were investigated by flow cytometry using DCFH-DA and Rh123 staining. The protein expression of HO-1, Cyt C and cleaved Caspase-3 was detected by Western blot. Result: The results showed that paeooiflorin at different doses (5, 10, 20 μmol · L^-1) could increase the cell viability and activities of antioxidant enzyme (GSH-Px, SOD and HO-1 ), prevent LDH release and lipid peroxidation (MDA) production, decrease the level of intracellular ROS, increase MMP, inhibit the release of cytochrome c from the mitochondria and attenuate activation of Caspase-3. Conclusion: Paeoniflorin can protect PCl2 cells from Aβ25-35 oxidative stress injuries. The mechanism may be related to its antioxidant action and inhibition of mitochondria-mediated caspase signaling path- ways.
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