Study On Regulation Of Leptin By Feeding Ob Gene Recombinant Yeast In Mice

With the increasing rapidly in obese patients,the number of patients with related diseases such as type 2 diabetes,atherosclerosis,cardiovascular disease and cancer is increasing,which makes obesity a major health problem covering the world.Studies have shown that the development of obesity is closely related to the out-of-control of the level of leptin encoded by the obesity gene(ob).The main physiological role of leptin is to control food intake and increase body energy expenditure.In view of this research,we began to explore the possibility of using leptin to treat obesity.However,the existence of leptin resistance makes this program ineffective in clinical treatment,and it only has an effect on a small number of obese individuals lacking leptin with obesity gene mutation.This phenomenon also provides a new way of thinking: through injection of neutral leptin antibody can react with circulating leptin in the blood,leptin antibody combines with circulating leptin in the blood,neutralizes and offsets the blood leptin,in turn,restores the sensitivity of leptin,allowing leptin in normal obese patients to perform their normal physiological functions.Saccharomyces cerevisiae can stimulate the immune response of dendritic cells(DCs).After that,immunotherapies developed around the interaction of dendritic cells and yeast gradually attracted the attention of the world.From the initial protein expression system to the subsequent surface display system,and then to the subsequent DNA / m RNA / sh RNA and other nucleic acid systems,the immune effect of this system is gradually optimized and enhanced.In this study,the ob gene was combined with the recombinant yeast system to develop an ob gene recombinant yeast against leptin.And using animal experiments to verify the effect of the feeding ob gene recombinant yeast under different dietary conditions(high fat diet,normal diet).The main research results of this article are as follows:1.Construct two of ob gene recombinant yeast with different effects: JMB84-CMV-OVA-ob and JMB84-CMV-ob.In this study,three different eukaryotic expression plasmids were constructed through gene cloning other techniques,and the successful expression of the plasmid JMB84-CMV-OVA-ob-T2A-GFP-poly A expression cassette was verified at the cellular level.Afterwards,based on the JMY1 yeast,two ob gene recombinant yeast systems with different effects were successfully constructed.2.Prokaryotic expression vector of ob gene was constructed to induce prokaryotic expression of ob gene,and recombinant leptin protein purification of prokaryotic expression of ob gene was used as antigen in subsequent antibody detection experiments.3.Evaluation of the performance of the ob gene recombinant yeast system in vivo.In this study,feeding ob gene recombinant yeast experiment under different feeding conditions(high fat diet,normal diet),using Western blot experiments,enzyme-linked immunosorbent assay and other experimental methods to verify the leptin antibody of different experimental groups,and screen out the recombinant yeast with better effect.In addition,through experimental testing,it was found that the recombinant yeast system can reduce levels of circulating leptin,total cholesterol,triglycerides in the blood,and can also effectively reduce the feed intake of experimental animals and reduce body weight gain.Especially for mice induced by a high-fat diet,oral administration of the system can significantly reduce the size of lipid droplets in liver tissue fatty lesions.In summary,this study successfully constructed two ob gene recombinant systems against leptin: JMB84-CMV-OVA-ob and JMB84-CMV-ob.Subsequently,we used the recombinant yeast system to achieve regulation of the phenomenon of leptin resistance,and evaluated the effects of the two different recombination yeast systems.And then we selected the recombination yeast system with better effect JMB84-CMV-OVA-ob,which can effectively reduce blood leptin and triglyceride,total cholesterol,food intake,weight gain reduction,and effective reduce the size of lipid droplets in fatty lesions of liver tissue also provide a useful reference for the future development of therapies using leptin resistance to treat obesity.