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Purple sweet potato pigments scavenge ROS, reduce p53 and modulate Bcl-2/Bax to inhibit irradiation-induced apoptosis in murine thymocytes

机译:紫色番薯色素清除ROS,降低p53并调节Bcl-2 / Bax抑制辐射诱导的鼠胸腺细胞凋亡

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Purple sweet potato (PSP) pigments were proved to protect murine thymocytes from ~(60)Co γ-ray-induced mitochondria-mediated apoptosis in our previous study. In this study, we further investigated the effect of PSP pigments on apoptosis related ROS, p53 and Bcl-2 family. Methods: Cell viability was analyzed by MTT. Apoptosis was certified by DNA ladder detection. Reactive oxygen species (ROS) were detected using 2',7',- dichlorofluorescein diacetate (DCFH-DA) probe. P53, Bcl-2 and Bax proteins were analyzed by western blot. The activities of caspase-3 and caspase-9 were determined by fluorogenic substrates detection. Results: PSP pigments treatment prior to 4Gy ~(60)Co γ-ray irradiation increased the cell viability and decrease the apoptosis. In the presence of PSP pigments, ROS was scavenged and followed by a p53-depression. A shift in Bcl-2/Bax ratio towards anti-apoptosis was observed as a result of p53-depression. The activities of caspase-9 and caspase-3 were reduced by PSP pigments pretreatment. Conclusions: PSP pigments have a cytoprotective activity against γ radiation. The protective effect of PSP pigments may be involving ROS scavenging, p53 depression and Bcl-2/Bax modulation in a caspase-dependent mitochondrial way.
机译:在我们先前的研究中,紫色甘薯(PSP)色素被证明可以保护鼠胸腺细胞免受〜(60)Coγ射线诱导的线粒体介导的细胞凋亡。在这项研究中,我们进一步研究了PSP色素对凋亡相关ROS,p53和Bcl-2家族的影响。方法:MTT法检测细胞活力。细胞凋亡通过DNA阶梯检测得到证实。使用2',7',-二氯荧光素二乙酸酯(DCFH-DA)探针检测活性氧(ROS)。通过蛋白质印迹分析P53,Bcl-2和Bax蛋白。通过荧光底物检测来确定caspase-3和caspase-9的活性。结果:在4Gy〜(60)Coγ射线辐照之前对PSP颜料进行处理可提高细胞活力并减少细胞凋亡。在PSP颜料存在下,清除ROS,然后进行p53抑制。由于p53抑制,观察到Bcl-2 / Bax比值朝着凋亡的方向移动。 PSP颜料预处理可降低caspase-9和caspase-3的活性。结论:PSP颜料对γ射线具有细胞保护活性。 PSP颜料的保护作用可能涉及以半胱天冬酶依赖性线粒体方式清除ROS,p53抑制和Bcl-2 / Bax调节。

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