The Effect Of Liraglutide On The Expression Of Type â…¢ Adenylate Cyclase And Its Role In Insulin Resistance And Obesity

objective:1.To investigate the effects of liraglutide on expression of adenylyl cyclase3in C57BL rat and diabetes rat.2.To investigate the effect of liraglutide on the levels of HOMA-IR and the adenylyl cyclase3in type2diabetes with or without obesity.3.To investigate the effects of liraglutide on expression of adenylyl cyclase3in human umbilical vein endothelial cells.Methods:1.Animal experiment:select twenty-fourC57BL/6J mice and sixteen db/db gene (obese rat model of type2diabetes). C57BL/6J mice were randomly divided into3groups (C57normal diet group, C57high fat feeding without intervention group and C57high fat feeding intervention group), with8rats in each group, and each experimental group of male and female pair:two experimental group C57BL/6J mice fed a high fat diet (34.9g%containing fat and protein26.2g%) for the C57high lipid intervention group and C57fat non-intervention group, in another experiment groups of C57BL mice were given normal diet (4.5g%fat and14.5g%protein) for C57material.16db/db mice, divided into2groups (DB intervention group and without DB intervention group), with8rats in each group, and each experimental group of male and female mating. Weekly measurements of body weight and tail vein blood glucose. From the beginning of the twelfth week, C57high-fat intervention group and DB intervention group mice were given subcutaneous injection of liraglutide100μg/kg, two times a day, the other three groups received normal saline as a control,12weeks after removal of eyeball mining angular vein blood, quickly remove the fat, liver, muscle, large intestine joint tissue, expression of TaqMan Realtime PCR for the detection of cell AC3mRNA expression and protein immunoblotting detection of Western in blot cells AC3protein. Venous blood centrifugation supernatant insulin detection.2.Clinical part:according to the standard of selecting32cases in NGT group and T2DM group25cases, according to the body mass index (BMI) were divided into2subgroups, BMI≥28Kg/m2as obesity group;18.5Kg/m2=BMI<24Kg/m2for non obese group. NGT obesity group15cases, male8cases, female7cases, NGT non obese group17cases, male9cases, female8cases; T2DM obesity group12cases, male6cases, female6cases, T2DM non obese group13cases, male7cases, female6cases. T2DM group received liraglutide0.6mg once daily, subcutaneous injection,1weeks after added to1.2mg, for16weeks, NGT obesity group were given health education. All of the subjects under fasting detection index, height (m), body weight (kg), body mass index (BMI), waist hip ratio(WHR), fasting blood glucose,HbA1C,HDL-C,TG,TC and LDL-C,AC3level,HOMA-IR; followed up every4weeks,a total of16weeks of follow-up. 3.HUVEC were cocultured with different concentration of liraglutide (0,10,100nmol/L) for8h or24h. Extraction of total RNA,AC3protein and mRNA expression in HUVECs were measured by Western bloting and reverse transcription polymerase chain reaction(RT-PCR)Results1. Animal parts:liraglutide significantly promote obese non-diabetic mice and obese the diabetic mice AC3mRNA and the protein expression,decrease weight,improve blood glucose(p<0.01). Liraglutide12weeks of treatment, db/db mice,with and without intervention group compared to the intervention group, weight,blood glucose decreased and insulin rises,the differences were statistically significant (P<0.05). No difference in C57BL/6J mice among the three groups glucose; C57high-fat intervention group and the C57normal diet group,body weight,insulin significantly increased,the difference was statistically significant (P<0.05);C57high-fat intervention group and high-fat without intervention group,body weight,insulin was significantly reduced,and the difference was statistically significant (P<0.05). Each group of mice fat cells, liver, skeletal muscle, the junction of large intestine and small intestine AC3mRNA expression comparison:db/db mice, the intervention group than non-intervention group AC3mRNA expression was significantly increased, and the differences were statistically significant (P<0.05). C57BL mice fat without intervention group AC3mRNA expression was significantly reduced, the difference was statistically significant (P<0.05); the high fat intervention group AC3mRNA expression was significantly increased statistically significant (P<0.05).2. Clinical part:(1) T2DM non-obese group:FPG,HbA1C,HOMA-IR and insulin levels of liraglutide after16weeks of treatment was statistically significant (P<0.05),HDL-C,AC3significant increased(P<0.05).(2) T2DM obese group: weight,BMI,WHR,FPG,HbA1C,insulin,HOMA-IR value liraglutide after treatment was significantly lower (P<0.05), TC, HDL-C, AC3higher than before treatment (respectively P<0.05, P<0.01).(3) T2DM obesity group, body weight, BMI, WHR decline than non-obese group was significantly (respectively, P<0.01, P<0.05), AC3increased there was a significant (P<0.01).(4) Statistical analysis showed that:the AC3with weight, BMI, WHR, of HOMA-IR was negatively correlated (r=-0.254,respectively,P<0.01, r=-0.228, P<0.05, r=-0.703, P<0.01,r=-0.212,P<0.05).Liraglutide treatment effect,FPG decreased pre-treatment FPG, HbA1C,HOMA-IR was positively correlated (r=0.914, respectively, P<0.01, r=0.771, P<0.01, r=0.194, P<0.05), Stepwise multiple stepwise regression analysis shows the AC3elevated treatment before treatment before weight, BMI, HOMA-IR was negatively correlated with pre-treatment AC3level of positive correlation; AC3for the dependent variable, body weight, BMI,WHR, HOMA-IR, FPG HbA1C, TG, TC, HDL-C and LDL-C as the independent variable stepwise multiple regression analysis, the results showed that body weight,BMI,WHR,HOMA-IR last into the regression equation, equation corrected r2=0.542, P=0.001.3.Liraglutide(10nmol/l) intervention in8hours,24hours AC3mRNA were significant (P<0.01),the endothelial cells expression in an upward trend with time (P<0.01).Liraglutide and HUVECs were cultured for24hours,with the liraglutide concentration (0,10,100nmol/L) increased, AC3mRNA expression levels were significantly increased (P<0.01) and protein expression levels were significantly increased (P<0.01),while NaN3has no effect on the expression of the AC3.Conclusions1.AC3was negatively correlated with body weight, BMI, WHR, HOMA-IR, liraglutide treatment can increase the AC3level, whether animal or in patients with high body weight and insulin resistance can be reversed by liraglutide intervention.2.Effects of liraglutide on expression of HUVECs in AC3display with the concentration of liraglutide and prolong the time of increased expression of AC3.