Preparation Of Strain DNZY1 Biosynthesis Lipopolysaccharide And Its Oral Sustained Release Microspheres

Diabetes is a common disease caused by genetic and environmental factors.High blood glucose was as the main clinical sign.The main symptoms were polydipsia,polyyuria,polyphagia and weight loss.The main method of treat diabetes was injecting insulin,insulin was the only hypoglycemic hormone that secreted by pancreatic B cells.However,the clinical treatment of diabetes has following problems:insulin can cause lower blood glucose that would lead life-threatening;the action time of insulin was too short,serious patients need inject 3-4 times daily;insulin as a treatment of diabetes,lower blood glucose,weight gaining,toxic and side effects such as cardiovascular disease would be arosed.Therefore,new antidiabetic drugs which had lower toxic and side effects,even non-toxic,better hypoglycemic effects would be studied,to improve inadequate treatment of diabetes.The predecessor of FG was FGF-21,has a long-term mechanism of action.During treat diabetes hypoglycemia does not be produced,at the same time regulating glucose and lipid metabolism,also improving insulin sensitivity.However,FG was unstable,easily degraded in vivo,and has potential immunogenicity.It was ineffective in the early stage of diabetes treatment.The key issue was how to improve stability of FG,and extend half-period of FG in vivo.In this study,the gene ebgineering strain DNZY1 produce FG was fermentated,and the metabolite was puritied.The purity and biological activity of FG were analysised by Western blotting.In addition,in order to solve FG unstable,degradable,potentially immunogenic,and ineffective in the early stage of diabetes treatment.The FG was wrapped by biodegradable material N-2-HACC/CMC that own syntheted.The physicochemical characters,stability,immune effect of the FG-N-2-HACC/CMC-MPs and the cell toxicity of the N-2-HACC/CMC were evaluated.The results demonstrated that:1)The FG produced by strain DNZY1 was obtained;2)The result of Western blotting analysis demonstrated that the reaction of FG produced by strain DNZY1 was specificity,and FG can react with anibody;3)The result of biological activity test demonstrated that the product obtained by strain DNZY1 has hypoglycemic function;4)The cytotoxicity of N-2-HACC/CMC was lower,the cell viability was(94.76±2.27)%(n=3);5)The optimal conditions of FG-N-2-HACC/CMC-MPs were obtained.The conditions were N-2-HACC concentration 2.0mg/mL,CMC concentration 1.2mg/mL,N-2-HACC:FG(V:V),stiring time 50min,stiring speed 1000r/min.The results demonstrated that the FG-N-2-HACC/CMC-MPs had been produced with morphous regulation,integrated profile,smooth surface,suitable size.The average particle size of the FG-N-2-HACC/CMC-MPs was 568.lnm with polydispersity index 0.331 and proper Zata Potential 26.45mV.The entrapment efficiency was(89.37±2.78)%(n=3),the loading capacity was(48.75±1.41)%(n=3);6)The release behavior of FG-N-2-HACC/CMC-MPs test in vitro demonstrated that FG-N-2-HACC/CMC-MPs release was a continuous release process after initial burst release;7)The FG-N-2-HACC/CMC-MPs has biological activity in vivo;8)The result of safety test demonstrated that FG-N-2-HACC/CMC-MPs has higher safety;9)The releasing test has shown that FG-N-2-HACC/CMC-MPs could make FG sustained release,the sffects were batter than FG;10)The result of permeability demonstrated that the route of administration of FG-N-2-HACC/CMC-MPs prepared in test was changed,still has hypoglycemic function,the experimental evidence of transform injiection to oral was provided.FG-N-2-HACC/CMC-MPs had been assembled successfully in this study,and had implemented a mechanism of the long-term mechanism of action in vivo,which could protect protein form degradation,increase the vaccination methods,decrease the vaccination times,enhance the immune effect.At the same time,the results also provided theoretical reference for developing a safe,efficient and better performance new diabetes drug.