The Study Of PPARγ Regulation On Adiponectin Secretion By RUVBL2

Adiponectin is an important adipokine found in recent years, which plays an important role in mediating lipid and glucose metabolism. It can stimulate fatty acid oxidation, inhibit hepatic gluconeogenesis and enhance insulin sensitivity. New functions also have been found about its regulation on the pathology of chronic inflammation, anti-atherosclerosis, and on food intake and weight via the nervous system. At present, there are a lot of reports about the transcriptional regulation of adiponectin as well as its post-translational modification. This study aims at getting further understand about the later stage of adiponectin secretion through studying protein RUVB2 which is involved in vesicle fusion.We hope to explain why the circular adiponectin levels were reduced in obese individuals from the aspect of vesicle fusion.In this study, preadipocyte cell line 3T3-L1 induced to differentiate and cell line HEK293 are used as our cell model. Quantitative real-time PCR(QRT-PCR), Chromonsome Immunoprecipitation(Ch IP),cell transfection and other molecular biological techniques were used to find out how PPARγ reglutes the process of vesicle fusion of the adiponectin to improve the screction of the adiponectin via enhancing the expression of the RUVBL2.The results are as follows:1.By RT-PCR and Western Blot,we find the similarity between the expression of the PPARγ and RUVBL2 during the differentiation.2. Bioinformatic analyzes the putative PPRE sites in RUVBL2 promoter. Then Luciferase reporter assay in HEK293 cells and Ch IP of the mouse white adipose tiusse comfirm the specific site where PPARγ bind to directly to promotes RUVBL2 m RNA level.3.Through Gene knockout or PPARγ agonist/inhibitor stimulation treatment experiments,we identified that the decrease of RUVBL2 inhibits adipoection screction,while changing PPARγ expression inhibits the RUVBL2 expression and adiponection screction.4 Though Gene overexpression experiments,we identified that the increase of RUVBL2 promotes adipoection screction,meanwhile the increase PPARexpression can enhance the adiponection screction.