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线粒体ROS改变参与石蒜碱诱导的HepG2细胞凋亡 被引量:1
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作者 抗晶晶 刘晓宁 《南京师大学报(自然科学版)》 CAS CSCD 北大核心 2019年第4期91-96,共6页
探究石蒜碱诱导HepG2细胞凋亡的分子机理. CCK-8实验检测细胞增殖情况;流式细胞术检测加或不加SS31情况下,石蒜碱对HepG2细胞凋亡比例的影响;用核酸染料Hoechst 33342评估凋亡细胞细胞核的形态变化;用Western blot分析技术检测PARP片段... 探究石蒜碱诱导HepG2细胞凋亡的分子机理. CCK-8实验检测细胞增殖情况;流式细胞术检测加或不加SS31情况下,石蒜碱对HepG2细胞凋亡比例的影响;用核酸染料Hoechst 33342评估凋亡细胞细胞核的形态变化;用Western blot分析技术检测PARP片段的表达情况.成功构建靶向线粒体的Mito-Grx1-roGFP2转基因稳定株肝癌细胞系HepG2. SS31作为线粒体ROS的定向清除剂,能够改善石蒜碱引起的HepG2细胞形态学损伤及细胞核内PARP切割,抑制石蒜碱诱导的细胞凋亡,降低反映线粒体ROS含量的405 nm/488 nm荧光比值.研究结果提示,线粒体内ROS水平的改变是石蒜碱诱导HepG2细胞凋亡的机制之一.石蒜碱有望成为治疗肝癌的潜力药物,值得更深入的研究. 展开更多
关键词 石蒜碱 人类肝癌HepG2细胞 细胞凋亡 mito-grx1-rogfp2 线粒体ROS SS31
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hpRNA的制备新方法及其在RNAi载体中的应用
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作者 刘晓宁 《安徽农业科学》 CAS 2016年第4期175-177,244,共4页
[目的]验证制备新方法所获得的稳定hpRNA是否可用于RNAi载体的构建并达到基因表达干扰的效果。[方法]利用制备新方法(2μL DNA与8μL H_2O混匀,95℃5 min,37℃15 min)制备hpRNA,后采用REGS方法构建Zm MDAR RNAi载体,通过Grx1-ro GFP2和... [目的]验证制备新方法所获得的稳定hpRNA是否可用于RNAi载体的构建并达到基因表达干扰的效果。[方法]利用制备新方法(2μL DNA与8μL H_2O混匀,95℃5 min,37℃15 min)制备hpRNA,后采用REGS方法构建Zm MDAR RNAi载体,通过Grx1-ro GFP2和荧光实时定量PCR检测基因表达干扰的效果。[结果]利用制备新方法制备50 nt hpRNA。Zm MDAR基因的表达量变化和Grxl-ro GFP2探针的荧光比值变化表明hpRNA介导的干扰载体引起了原生质体氧化还原状态的变化。[结论]通过制备新方法所制备的hpRNA为RNAi载体所介导的干扰作用奠定基础。 展开更多
关键词 发夹RNA REGS ZmMDAR RNAI Grx1-rogfp2
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Exploring the precision redox map during fasting-refeeding and satiation in C.elegans
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作者 Xinhua Qiao Lu Kang +7 位作者 Chang Shi Aojun Ye Dongli Wu Yuyunfei Huang Minghao Deng Jiarui Wang Yuzheng Zhao Chang Chen 《Stress Biology》 2023年第1期187-200,共14页
Fasting is a popular dietary strategy because it grants numerous advantages,and redox regulation is one mecha-nism involved.However,the precise redox changes with respect to the redox species,organelles and tissues re... Fasting is a popular dietary strategy because it grants numerous advantages,and redox regulation is one mecha-nism involved.However,the precise redox changes with respect to the redox species,organelles and tissues remain unclear,which hinders the understanding of the metabolic mechanism,and exploring the precision redox map under various dietary statuses is of great significance.Twelve redox-sensitive C.elegans strains stably expressing genetically encoded redox fluorescent probes(Hyperion sensing H_(2)O_(2) and Grx1-roGFP2 sensing GSH/GSSG)in three organelles(cytoplasm,mitochondria and endoplasmic reticulum(ER))were constructed in two tissues(body wall muscle and neurons)and were confirmed to respond to redox challenge.The H_(2)O_(2) and GSSG/GSH redox changes in two tissues and three organelles were obtained by confocal microscopy during fasting,refeeding,and satiation.We found that under fasting condition,H_(2)O_(2) decreased in most compartments,except for an increase in mitochondria,while GSSG/GSH increased in the cytoplasm of body muscle and the ER of neurons.After refeeding,the redox changes in H_(2)O_(2) and GSSG/GSH caused by fasting were reversed in most organelles of the body wall muscle and neurons.In the sati-ated state,H_(2)O_(2) increased markedly in the cytoplasm,mitochondria and ER of muscle and the ER of neurons,while GSSG/GSH exhibited no change in most organelles of the two tissues except for an increase in the ER of muscle.Our study systematically and precisely presents the redox characteristics under different dietary states in living animals and provides a basis for further investigating the redox mechanism in metabolism and optimizing dietary guidance. 展开更多
关键词 Redox Fasting Refeeding Satiation Hyperion Grx1-rogfp2 Caenorhabditis elegans
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