Therapeutic Effects And Mechanisms Of CTRP1 In Inhibiting Obesity And Improving Glucose Homeostasis In Mice

Experimental background and purpose:In recent decades,obesity and other metabolic diseases have increasingly affected people’s physical and mental health,so it is particularly important for the treatment of obesity.At present,comprehensive treatment based on behavior and diet is mostly used for obesity treatment,but the effect is poor.More severe obese patients are treated with drugs.Central drugs(sibutramine)and non-central drugs(orlistat)will cause strong side effects and the effect is not significant.In recent years,researchers have been more inclined to find a drug with significant effects and no obvious side effects.Compared with chemical drugs,biological drugs are more in line with people’s expectations.CTRP1 protein is an adipose factor secreted by adipose tissue and with a molecular weight of 35 KDa.It is a member of the CTRP family.Existing literatures report that CTRP1 is associated with coronary heart disease,dehydration and hypotension,and chondrocyte proliferation and maturation,Inflammation,and glucose homeostasis.Jonathan and his colleagues showed that CTRP1 can control obesity induced by high-fat food.This effect is partly due to CTRP1 activating the AMPK pathway in muscle tissue,inhibiting the activity of ACC and improving fatty acid oxidation.Overall,the role of CTRP1 in metabolic homeostasis and energy balance is not clear.We studied the role and possible mechanisms of CTRP1 in metabolic homeostasis and energy balance.Experimental method:1)Plasmid of p LIVE-CTRP1 was using molecular cloning techniques such as PCR,enzyme digestion,enzyme ligation,and transformation.The p LIVE-SEAP plasmid is an existing plasmid in the laboratory.After the sequencing is correct,we use the endotoxin-free plasmid mass extraction kit to extract a large number of plasmids for use.2)Using the hydrodynamic gene delivery technology,the plasmid carrying the CTRP1 gene is delivered to the mouse body,and sustained high expression in the liver and secretion into the blood circulation.3)Using high-fat diet-induced obesity models to study the effects of CTRP1 on HFDinduced obesity,hyperglycemia,insulin resistance,and liver lipid accumulation.The impacts of CTRP1 on metabolic homeostasis of obese and diabetic mice were examined using HFD-induced obese and STZ-induced diabetic models4)Using Western Blot and Real Time PCR technology to analyze changes in genes related to liver,adipose tissue pathway proteins,lipid metabolism,glucose metabolism,inflammation,and energy metabolism.Metabolic cages were used to monitor changes in mouse activity,food consumption,and energy expenditureExperimental results:1)The p LIVE-CTRP1 recombinant plasmid was successfully constructed and a high expression system in three animal models was successfully established.2)CTRP1 gene transfer can inhibit obesity,hyperglycemia,insulin resistance and fatty liver caused by HFD.In obese and STZ-induced diabetic mice,CTRP1 improves glucose homeostasis.3)Mechanism research shows that CTRP1 gene trasfer increases the activity of mice,up-regulates the expression of genes related to thermoregulation in brown adipose tissue,and downregulates the expression of genes connected to fat metabolism and glucose homeostasis in liver tissue and inflammation-related genes in white adipose tissue.High expression of CTRP1 also activated AMPK and PI3 K / AKT signaling pathways.Experimental conclusions:These phenomena reveal that CTRP1 plays an critical role in managing metabolic homeostasis and energy metabolism,and can be used as a target for drug intervention in the treatment of metabolic diseases.