Study On The Mechanism Of Intestinal Flora To Reduce Weight And Lipid Of COST And Preparation Of Enteric-coated Capsule

Obesity is a chronic endocrine malady,closely associated with metabolic diseases like polygenic disease,non-alcoholic liver disease,hyperlipoidaemia,and coronary-artery disease.Currently,the sole over-the-counter anti-obesity drug approved by FDA is Orlistat,but it has been warned by the FDA for liver and kidney toxicit.Therefore,there’s associate pressing ought to notice a secure and effective drug to combat the event of obesity.COST(Chito-oligosaccharide with average Mw≤ 1000 Da,)is a natural product formed by the combination of glucosamine and N-acetyl glucosamine.Our analysis cluster has been committed to the analysis on the lipid-lowering function of COST for a long time.Early experiments show that COST has a good anti-obesity effect and effectively regulates the intestinal microbes of obese animal models.Therefore,this project takes the gut microbes after COST intervention as the analysis target,adopts the establishment of pseudo-sterile mouse models,explores the lipid-lowering activity and pharmacological mechanism of gut microbes after COST intervention,and further studies the weight loss and lipid-lowering mechanism of COST.Use antibiotics to build pseudo-sterile mouse models.After feeding C57BL/6J mice on a high-fat and high-sugar diet with antibiotic drinking water for 28 days,the diversity and abundance of microbes in the intestines of the mice were significantly reduced,and the proportion of microbes at each classification level also changed significantly.It shows that this method can successfully induce the establishment of a pseudo-sterile mouse model,and provide a good environment for the colony transplant recipients for subsequent fecal microbial transplantation(FMT)experiments.The results of the FMT experimental activity study show that the microorganisms after COST intervention have a good lipid metabolism regulation activity.In an environment induced by a high-fat and high-sugar diet,the gut microbes after COST intervention can reduce the weight of mice,effectively inhibit the overgrowth and expansion of adipose tissue,regulate the four levels of blood lipids,reduce the level of lipid metabolites in mice,and maintain normal blood sugar level,lower liver function indicators,have a certain effect of lowering blood lipids and blood sugar.The analysis results of this mechanism show that COST can regulate the disorder of lipide metabolism within the body by rising the enteral microecological balance.Using 16 S r DNA amplicon sequencing to examine the intestinal microbes of mice,the gut microbes after COST intervention can maintain a higher number of OTUs and a lower ratio of Firmicutes/Bacteroidetes after transplantation,with higher species diversity and species abundance.Further analysis shows that COST can effectively regulate anti-obesity-related intestinal microbes,such as Clostridium,Dehalobacterium,Adlercreutzia,Akkermansia,Streptococcus,and Dehalobacteriaceae.Besides,RT-PCR results show that the microorganisms after COST intervention can effectively reduce the de novo synthesis of lipids in the liver,promote lipid β-oxidation degradation,and promote the conversion between cholesterol and bile acids.Activity and mechanism studies have shown that COST can play a good anti-obesity effect by improving intestinal microbes.FMT results showed that COST could transfer lipid-lowering activity through intestinal microorganisms.According to its mechanism of action,COST was developed into an enteric-coated capsule.A single factor test is used to determine the best preparation process.According to the optimal prescription,Eudragit L100 is used as the enteric matrix,HPMC is used as the content release rate regulator,and the DRcaps enteric capsule shell is used as the capsule shell.COST enteric-coated capsules were prepared according to the ratio of Eudragit L100:HPMC: COST = 4.0:1.0:1.0.The prescription verification experiment proved that the prescription process is reasonable and stable.The average COST content of enteric-coated capsules is 16.66%,the average moisture content is6.16%,and the average disintegration time is 8 minutes.The difference in filling volume and capsule appearance conform to the relevant requirements of the Chinese Pharmacopoeia(2020 edition).COST enteric-coated capsules remain intact for 2 hours in acidic artificial gastric juice;however,they can be completely dissolved and released within 120 minutes in an alkaline artificial intestinal juice environment,indicating that the preparation of enteric-coated capsules complies with relevant regulations.In summary,based on the intestinal flora,this article studied the lipid-lowering activity and mechanism pathways of the intestinal flora after COST intervention through a pseudo-sterile mouse model.The results show that the lipid-lowering function of COST can be transferred through the intestinal microorganism as a medium.After the intervention,the intestinal flora can show good weight loss and lipid-lowering functions.It is speculated that COST can achieve the treatment of obesity by improving the obesity-related flora.According to the characteristics of COST’s mechanism of action,it is prepared into an enteric-coated capsule product that meets the relevant requirements of the Chinese Pharmacopoeia(2020 Edition),in order to develop a safe and effective lipid-lowering five-category Chinese medicine natural medicine to enrich the relatively vacant anti-obesity the drug market.