线粒体温和解偶联剂的发现及作用机制研究

发布时间：2018-03-22 22:14

本文选题：线粒体　切入点：膜电位　出处：《华东师范大学》2011年硕士论文　论文类型：学位论文

【摘要】：线粒体是真核细胞中最重要的细胞器之一,除了为细胞提供能量,还参与细胞信号转导、分化与生长、凋亡等生命过程。许多疾病的发生与线粒体功能失常密切相关,包括神经退行性疾病、糖尿病、肥胖、肿瘤等。近年来,对线粒体功能的研究日益为研究人员所关注。因此,我们希望通过寻找靶向线粒体功能的温和调节剂而获得对糖脂代谢有改善作用的先导化合物。线粒体膜电位是线粒体功能的一个重要指针,我们建立L6肌细胞线粒体膜电位调节剂高通量筛选模型,对国家新药筛选中心小分子化合物库近4000样次化合物进行随机筛选,得到了一个低毒并且能够浓度依赖的降低线粒体膜电位的小分子化合物LGH00272。LGH00272可以比较温和地降低L6肌细胞的线粒体膜电位,并且温和的促进L6肌细胞、大鼠原代肝细胞和分离的大鼠肝脏线粒体耗氧,同时对细胞中ATP的合成无明显抑制,对乳酸的生成有轻微促进。进一步在代谢性细胞和整体动物水平上评价该化合物的药效表明：LGH00272能够促进L6肌细胞的葡萄糖吸收和脂肪酸氧化,降低HepG2肝癌细胞甘油三酯和胆固醇含量。在整体动物实验中,LGH00272长期给药能够使饮食诱导肥胖(diet induced obesity, DIO)小鼠的体重增长更加缓慢,并且对摄食没有影响。LGH00272还可以降低DIO小鼠血糖和胆固醇,腹腔注射葡萄糖耐量实验也进一步证明LGH00272能够提高DIO小鼠的糖耐受力。此外,LGH00272对糖异生也有明显抑制,不仅能够抑制大鼠原代肝细胞的糖输出,还能降低长期给药DIO小鼠肝脏中糖异生关键基因的表达水平。LGH00272还降低了肝脏和脂肪比重,同时降低脂合成及增加脂代谢相关基因的表达。综上所述,我们通过线粒体膜电位高通量筛选得到了一个温和的线粒体解偶联剂,该化合物在细胞水平和整体动物水平对糖脂代谢均有明显的改善功能,结果表明通过寻找温和的线粒体解偶联剂可以获得新颖的治疗代谢综合症的先导化合物,对进一步阐明线粒体和代谢综合症的关系起到积极的意义。
[Abstract]:Mitochondria is one of the most important organelles in eukaryotic cells. In addition to providing energy to the cells, mitochondria are also involved in cell signal transduction, differentiation and growth, apoptosis and other life processes. Many diseases are closely related to mitochondrial dysfunction. These include neurodegenerative diseases, diabetes, obesity, tumours, and so on. In recent years, the study of mitochondrial function has attracted more and more attention. We hope to obtain lead compounds that improve glucose and lipid metabolism by looking for mild regulators targeting mitochondrial function. Mitochondrial membrane potential is an important pointer to mitochondrial function. We established a high-throughput screening model for mitochondrial membrane potential modulators in L6 myocytes and randomly screened nearly 4000 small molecular compounds in the small molecular compound library of the National Center for New Drug screening. A low toxic and concentration-dependent low molecular compound, LGH00272.LGH00272, could decrease the mitochondrial membrane potential of L6 myocytes mildly, and promote L6 myocytes mildly. The primary rat hepatocytes and isolated rat liver mitochondria consumed oxygen, while the synthesis of ATP in the cells was not significantly inhibited. Further evaluation of the drug efficacy of the compound at the metabolic and overall animal levels showed that: LGH00272 promoted glucose absorption and fatty acid oxidation in L6 myocytes. Reduce the content of triglyceride and cholesterol in HepG2 hepatoma cells. In the whole animal experiment, the long-term administration of LGH00272 could slow the weight gain of diet-induced obese induced mice. LGH00272 also decreased blood glucose and cholesterol in DIO mice, and glucose tolerance test by intraperitoneal injection showed that LGH00272 could increase glucose tolerance in DIO mice. LGH00272 also inhibited glucose heterogenesis in DIO mice. It not only inhibited the glucose output of rat primary hepatocytes, but also decreased the expression level of glycosylated key genes in the liver of long-term DIO mice. LGH00272 also decreased the specific gravity of liver and fat. At the same time, it decreased lipid synthesis and increased the expression of genes related to lipid metabolism. In conclusion, we obtained a mild mtDNA uncoupling agent through high throughput screening of mitochondrial membrane potential, which can significantly improve glycolipid metabolism at cell level and animal level as a whole. The results show that a novel lead compound for the treatment of metabolic syndrome can be obtained by searching for mild mitochondrial uncoupling agents, which plays a positive role in further elucidating the relationship between mitochondria and metabolic syndrome.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R341

【引证文献】