秀丽新杆线虫肌管素磷酸酶生物学功能的研究及长期酒精刺激对线虫的影响

发布时间：2018-04-30 06:08

本文选题：秀丽新杆线虫 + 肌管素　；参考：《吉林大学》2011年博士论文

【摘要】：第一篇:蛋白质磷酸酶是细胞信号转导过程中发挥至关重要作用的一类蛋白质,其家族成员与多种人类疾病的发生、发展密切相关。肌管素磷酸酶亚家族(Myotubularins,MTMs)隶属于蛋白质磷酸酶超家族,该亚家族成员以磷酸肌醇(phosphatidylinositol, PI)作为其特异性的催化底物进行去磷酸化。人类的肌管素磷酸酶机组成员MTM-1突变可导致X-连锁的肌小管性肌病,MTMR2和MTMR13的突变可导致腓骨肌萎缩症,但肌管素磷酸酶引发其相关疾病的机理尚不清楚。本研究选用秀丽新杆线虫作为生物模型,利用RNA干扰的方法及相关的突变体对线虫肌管素磷酸酶成员ceMTM3、ceMTM6和ceMTM9进行了研究。我们的研究结果表明,ceMTM3能够影响线虫体内肌纤维的保持并对线虫体内自吞噬的发生起着负调控的作用。利用RNA干扰的方法降低线虫体内ceMTM3的表达量可加速线虫在衰老的过程中肌纤维的流失,使线虫的肌肉细胞发生萎缩变小,并在线虫体内肌肉细胞及其他组织中引起自吞噬水平显著升高。在自吞噬水平升高后,ceMTM3表达量的降低还导致了细胞体内溶酶体水平升高以及细胞坏死。Atg-18在线虫体内发生自吞噬的过程中能够与PI3P相结合并对自吞噬小体的形成起重要的调节作用,atg-18突变型线虫自身存在运动障碍,并且相比野生型线虫其肌肉细胞内肌纤维排列混乱。当对atg-18突变型线虫进行ceMTM3干扰后,我们发现在atg-18突变型线虫的体内降低ceMTM3的表达量并未使该突变株线虫体内的肌纤维排列进一步恶化。这说明ceMTM3对线虫肌纤维保持的调节作用是通过其对自吞噬的调控来实现的。本研究为揭示人类肌纤维保持的调节机制及治疗人骨骼肌减少症提供了依据和基础。此外,我们还对线虫体内另外两名肌管素成员ceMTM6和ceMTM9的生物学功能进行了研究。研究结果表明,mtm-6突变型线虫相比野生型体长较短、存在运动障碍。在发育的过程中进行肌肉染色的结果表明,相比野生型,mtm-6突变型的线虫肌细胞间隙较大且肌细胞较小,此外,约20% mtm-6突变型线虫的生殖腺在发育的过程中延伸方向发生了错误。整合素在细胞表面的分布和表达对细胞之间的粘附和细胞迁移有重要的调节作用。我们初步的研究结果表明,mtm-6能够影响线虫体内神经细胞DTCs表面整合素INA-1的表达和分布,而这极有可能是ceMTM6影响线虫肌肉细胞和生殖腺发育的原因之一。ceMTM9不具备催化活性中心,之前的研究结果表明,ceMTM9可与ceMTM6结合形成复合体共同发挥作用。本研究的结果表明,mtm-9突变型线虫可使线虫发育迟缓、体长较小、产卵数明显降低Bagged Worm比率显著升高并且寿命缩短。对mtm-9突变型线虫进行ceMTM3干扰的结果表明,ceMTM9和ceMTM3之间可能存在着相互代偿的作用。第二篇:酒精饮料是人类日常生活中必不可少的饮品。长期酗酒对人体有较大伤害,但近期的研究结果表明,长期适量饮酒可降低人类某些疾病的发病率,因此饮酒的利弊仍存在争议。本研究以秀丽新杆线虫作为模型,研究了不同浓度的酒精长期刺激线虫各种生命活动的影响。研究结果表明,长期高浓度的酒精刺激(≥4%)使线虫寿命缩短、产卵数减少、运动能力减弱。有趣的是,长期低浓度的酒精刺激(1-2%)可使线虫寿命延长。尽管产卵数有所减少,但长期2%浓度的酒精刺激可使线虫的寿命延长约2.76天,延长率达14.53%,并且伴随着运动的衰老程度减缓。研究发现,这种作用与线虫体内的类胰岛素信号通路以及转录因子Sir-2.1密切相关。相比未刺激的对照组,无论从卵、幼虫还是成虫开始2%酒精刺激都可使线虫寿命显著延长。本研究证明了线虫是研究长期酒精刺激的优秀模型并且长期少量的酒精刺激能够对机体产生积极的作用。本研究为进一步研究不同剂量酒精长期刺激对人类的生理功能的影响以及所引发的与之相关的分子机制奠定了基础。
[Abstract]:Protein phosphatase is a kind of protein that plays a vital role in cell signal transduction. Its family members are closely related to the development of a variety of human diseases. The Myotubularins (MTMs) belongs to the protein phosphatase superfamily, and the member of the subfamily is inositol phosphate (phosphatidylino). Sitol, PI) dephosphorylation as its specific catalytic substrate. Human myotin phosphatase unit MTM-1 mutation can lead to X- linked myomatosis. Mutations in MTMR2 and MTMR13 can lead to fibula muscular dystrophy, but the mechanism of myotin phosphatase that causes its related diseases is not clear. As a biological model, the ceMTM3, ceMTM6 and ceMTM9 of the nematode myotube phosphatase were studied by means of RNA interference and related mutants.
Our results showed that ceMTM3 could affect the retention of the muscle fibers in the nematode and negatively regulated the self phagocytosis of the nematode in the nematode. The reduction of the ceMTM3 expression in the nematode by RNA interference could accelerate the loss of muscle fibers in the aging process, and reduce the atrophy of the nematode muscle cells. The level of autophagy increased significantly in the muscle cells and other tissues in the body of the worm. The decrease of the expression of ceMTM3 also resulted in the increase of the lysosome level in the cells and the formation of the autophagy of the autophagocytic body in the process of the autophagy of the cell necrosis.Atg-18 in the body of the cells after the increase of autophagy. Important regulation, atg-18 mutant nematodes themselves have dyskinesia, and the muscle fibers in the muscle cells of the wild type nematode are confused. When ceMTM3 interference is done to atg-18 mutant nematodes, we found that the reduction of ceMTM3 in the body of the atg-18 mutant nematode did not make the muscle fiber in the mutant nematode. This shows that the regulation of ceMTM3 on the retention of nematode muscle fibers is achieved through its regulation of autophagy. This study provides a basis and basis for revealing the regulatory mechanism of human muscle fiber retention and the treatment of human skeletal myocytosis. In addition, we also have two other myosin members, ceMTM6 and C, in line worms. The biological function of eMTM9 was studied. The results showed that the mtm-6 mutated nematode was shorter than the wild type, and there was a dyskinesia. The results of muscle staining during development showed that the myocytes of the nematode of the mtm-6 mutant were larger and smaller than the wild type. In addition, about 20% mtm-6 mutant nematodes were born. The distribution and expression of integrin on cell surface have important regulatory effect on cell adhesion and cell migration. Our preliminary results show that mtm-6 can affect the expression and distribution of DTCs surface integrin INA-1 in the nerve cells of the nematode, and this is very likely. It is one of the reasons that ceMTM6 affects the development of the nematode muscle cells and the reproductive glands of the nematode..ceMTM9 does not have the catalytic activity center. The previous research results show that ceMTM9 can combine with ceMTM6 to form complex complex. The results of this study show that the mtm-9 mutant nematode can make the nematode grow slow, the body length is smaller, and the number of spawns obviously reduces the Bagged W. The ORM ratio was significantly higher and the life span was shortened. The results of ceMTM3 interference to the mtm-9 mutant nematode showed that there might be a mutual compensatory effect between ceMTM9 and ceMTM3.
Second: alcohol is an essential drink in human daily life. Long term alcohol abuse is very harmful to people. But recent research shows that long term alcohol consumption can reduce the incidence of certain diseases in human beings. Therefore, the advantages and disadvantages of drinking are still controversial. The effects of alcohol on the life of the nematodes for a long time. The results showed that long term high concentration of alcohol (> 4%) shortened the life span of the nematode, reduced the number of spawns, and weakened the exercise ability. Interestingly, the long term low concentration of alcohol stimulation (1-2%) could prolong the life of the nematode. Although the number of eggs was reduced, the long-term 2% concentration of alcohol was stimulated. The life span of the nematode can be extended by about 2.76 days, with an extension of 14.53%, and the slowing of the aging of the movement. The study found that this effect is closely related to the insulin like signaling pathway and the transcription factor Sir-2.1 in the nematode. Compared to the unstimulated control group, the 2% alcohol stimulation from the egg, the larva and the adult can make the nematode in the nematode. This study demonstrated that the nematode is an excellent model for long-term alcohol stimulation and a long amount of alcohol stimulation can have a positive effect on the body. This study provides a further study of the effects of the long-term stimulation of alcohol on the physiological functions of human beings and the related molecular mechanisms. Basics.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R341

【共引文献】