Correlation And Intervention Of Plasma RBP4 And Apelin12 With Diabetic Cardiomyopathy

Part I: Correlation between plasma RBP4,Ang?,insulin resistance and diabetic cardiomyopathy.Background Diabetes mellitus(DM)is a chronic metabolic disease based on insulin resistance and insulin secretion defects.Diabetes mellitus significantly increases the risk of cardiovascular disease and heart failure in patients and causes diabetic cardiomyopathy(DCM).Since 1972,Rubler et al.first proposed the concept of diabetic cardiomyopathy(DCM),and then paid more and more attention to DCM clinically.Diabetic cardiomyopathy showed obvious microvascular lesions,thickened vascular basement membrane,and deposition of Pas(glycogen staining)positive substances in myocardial stroma.As a special retinol carrier,RBP4(serum retinol-binding protein 4)can be secreted by fat cells,and there is increasing evidence that RBP4 is involved in the occurrence and development of obesity,insulin resistance and type 2 diabetes.This study included 92 patients with diabetic cardiomyopathy,only 145 cases the diabetes patients,control group 56 cases,in view of the three groups of patients serum RBP4,Ang?,Hs CRP,IL-10 levels,insulin resistance index,and study the relationship between them,for the pathogenesis of diabetic cardiomyopathy and treatment to provide more clinical data.Objective To observe the general data,biochemical indexes,fasting blood glucose,basal insulin level,glycosylated hemoglobin,homa-ir,Hs CRP,RBP4,il-10,Hs CRP,Ang? levels and comparative analysis of the three groups in diabetic cardiomyopathy patients,simple diabetes patients and the control group;Correlation analysis and single factor and multiple factor regression analysis were also performed.Materials and methods Patients admitted to the department of geriatrics and endocrinology of the first affiliated hospital of zhengzhou university from January 2015 to December 2016 who were initially diagnosed with "type 2 diabetes mellitus" in the department of geriatrics and endocrinology were collected.Patients included in the study did not take ACEI/ARB drugs,statins,diuretics orally or inject insulin.The selected patients included 145 patients with type 2 diabetes and 92 patients with diabetic cardiomyopathy.A total of 56 cases of healthy people in the physical examination center of the first affiliated hospital of zheng university were collected and those with normal glucose tolerance test and normal glycosylated hemoglobin were used as the control group.A total of 293 cases.All men in a signed written informed consent,accept routine blood examination,blood biochemical analysis,24 hours trace albumin detection,fundus examination,electrocardiogram,heart colour to exceed,and coronary angiography,etc.,improve the body mass index(BMI,waist-to-hip ratio WHR,systolic pressure,diastolic pressure,FPG SP,PPG,FINS,PINS,Hb A1 c,Homa IR,RBP4,Hs CRP,Ang?,IL-10 and related indicators.To the above data results,comparative analysis and correlation analysis.For the count data,mean + /-standard deviation(x + /-s)was used to indicate that,for the measurement data,t test was used to compare the two groups,which showed significant difference(P<0.05)and statistical significance.Pearson correlation analysis and Logistic univariate and multivariate regression analysis;SPSS was used for statistical analysis of 19.0 software version.Results 1.BMI,waist-hip ratio,Hb A1 C,TC,ldl-c and PPG of the DCM group and the DM group were significantly higher than those of the control group,and the incidence of atrial fibrillation in the DCM group was significantly higher than that of the DM group and the control group(P<0.05).DCM group HOMA-IR values compared with pure DM group and the control group with significant difference,the plasma of patients with DCM RBP4 concentration and the concentration of serum Hs CRP was significantly higher than that of DM group and the control group,P < 0.01,the plasma IL-10 in DCM group,simple diabetes levels were significantly lower than the control group comparison,DCM group,pure Ang? DM group than the control group comparison level significantly high,significant difference(P < 0.01).2.Serum RBP4 had a significant positive correlation with af history,BMI,waist-hip ratio,homa-ir,Hb A1 C,and ldl-c,and a negative correlation with FINS.After adjusting for age and gender,serum RBP4 is still significantly correlated with atrial fibrillation history,homa-ir,il-10 and Ang?.There was a significant positive correlation between serum Ang? and the history of atrial fibrillation,BMI,waist-hip ratio,SBP,homa-ir and RBP4.After adjusting for age and gender,there was still a significant correlation between serum Ang? and the history of atrial fibrillation,waist-hip ratio,SBP and homa-ir.According to univariate Logistic regression analysis,history of atrial fibrillation,waist-hip ratio,homa-ir,Ang?,RBP4 and other five factors have an impact on the prevalence of diabetic cardiomyopathy,and among these five variables,waist-hip ratio,Ang? and RBP4 have the largest impact on the formation and deterioration of diabetic cardiomyopathy and cardiac function.Conclusion 1.There is a significant correlation between RBP4 and Ang?,which promote and influence each other and obviously affect the occurrence and development of diabetic cardiomyopathy,which can be controlled as a high risk factor of diabetic cardiomyopathy.2.Multiple inflammatory factors and anti-inflammatory factors are involved in the occurrence and development of diabetic cardiomyopathy.3.Blood RBP4 may be a predictor of insulin resistance,type 2 diabetes and diabetic cardiomyopathy.Part II: Relationship between blood Apelin12,insulin resistance,Ang? and insufficiency of cardiac function in diabetic cardiomyopathy.Background Diabetes significantly increases the risk of cardiovascular disease and heart failure,known as diabetic cardiomyopathy(DCM).Left ventricular diastolic dysfunction(LVDD)is one of the earliest cardiac changes in patients with DCM.Myocardial fibrosis(MF)is the most harmful type of diabetes,with epidemiology showing that about 80% of diabetics end up dying of heart failure,arrhythmia and sudden death.Framingham's study found that the incidence of heart failure in men with diabetes was 3.8 times higher than in the control group and 5.6 times higher in women.When the diastolic function is damaged,the E/A ratio presents A reverse change,LVEDP(left ventricular final diastolic pressure)increases,LVEDV(end-diastolic volume)presents A normal or increased performance,and LVEDP/ LVEDV ratio increases.Nt-probnp is the brain natriuretic peptide precursor(pro-bnp)split after the production of inactive n-terminal fragment,and the degree of heart failure has a positive correlation.Apelin is a new member of RAS system.It is an endogenous ligand of angiotensin II type 1 receptor related protein APJ,and together with APJ,it participates in various pathophysiological mechanisms in vivo.This study included 92 patients with diabetic cardiomyopathy,simple diabetes group 145 cases,control group 56 cases,this study mainly Apelin-12 as object,study in patients with diabetic cardiomyopathy and Ang?,insulin resistance and left ventricular diastolic dysfunction,NT-pro BNP,the relationship between the pulmonary artery systolic blood pressure and other factors,and the factors influence on diabetic cardiomyopathy cardiac insufficiency,for the pathogenesis of diabetic cardiomyopathy and treatment to provide more clinical data.Objective Diabetic cardiomyopathy patients(mild heart failure group and severe heart failure group),simple diabetes patients and the control group,four groups of general data,biochemical indicators,fasting glucose,basic insulin level,glycosylated hemoglobin,homa-ir,Hs CRP,Apelin12,Ang?,homa-ir,LVDD,nt-probnp,PASP level and four groups of comparative analysis;Pearson correlation analysis and Logistic univariate and multivariate regression analysis were performed.Materials and methods Patients admitted to the department of geriatrics and endocrinology of the first affiliated hospital of zhengzhou university from January 2015 to December 2016 who were initially diagnosed with "type 2 diabetes mellitus" in the department of geriatrics and endocrinology were collected.Patients included in the study did not take ACEI/ARB drugs,statins,diuretics orally or inject insulin.The selected patients included 145 patients with type 2 diabetes and 92 patients with diabetic cardiomyopathy.A total of 56 cases of healthy people in the physical examination center of the first affiliated hospital of zheng university were collected and those with normal glucose tolerance test and normal glycosylated hemoglobin were used as the control group.A total of 293 cases.After the informed consent was signed by all the selected subjects,BMI,WHR,SP,diastolic blood pressure,FPG,PPG,FINS,PINS,Hb A1 c,Homa-IR,LVEDD(mm),LVEDP(mm Hg),pulmonary artery systolic blood pressure(PASP),ejection fraction(EF%),E/A value and other relevant indicators were improved.For the count data,mean + /-standard deviation(x + /-s)was used to indicate that,for the measurement data,t test was used to compare the two groups,which showed significant difference(P<0.05)and statistical significance.Pearson correlation analysis and Logistic univariate and multivariate regression analysis;SPSS was used for statistical analysis of 19.0 software version.Results 1.The BMI,waist-to-hip ratio,Hb A1 C,TC,LDL-C,and hypertension history of DCM group and DM group were significantly higher than those of the control group(P<0.05).Compared with DM alone,DCM group was more likely to produce house.The tremor phenomenon,between the DCM group and the control group(P < 0.05),the former had a higher probability of atrial fibrillation.The FINS of the DM group and the FINS of the control group(P<0.05)were higher than those of the DCM group.The FINS level of the diabetic group was higher than that of the control group,and there was no significant difference between the two groups.HOMA-IR,NT-pro BNP,LVEDD,LVEF%,and Ang? in the DCM group were significantly higher than those in the simple diabetes group,which was also higher than the control group,but the E/ of the DCM group.The A value was significantly lower than the latter two,and P<0.01;the severe heart failure group in DCM had more significant increase in HOMA-IR,NT-pro BNP,LVEDD,LVEF%,PSAP,Ang? than the mild heart failure group.In the DCM,the severe heart failure group had lower E/A value than the mild heart failure group,P<0.05;the DCM mild heart failure group(NYHA class I,II)Apelin12 increased significantly,and the Apelin12 level in the DCM severe heart.The decline group showed a decline.Compared with DCM mild heart failure group,there were statistical differences in HOMA-IR,NT-pro BNP and LVEDD in the severe heart failure group(P<0.05).Pearson correlation analysis showed that the serum Apelin12 and BMI,homa-ir,E/A value,nt-probnp,atrial fibrillation history,Ang? showed A significant correlation,adjusted for age and gender,serum Apelin12 and homa-ir,E/A value,nt-probnp,Ang? still have A significant correlation.According to univariate Logistic regression analysis,BMI,homa-ir,E/A value,Apelin12,Ang? and other 5 factors have an impact on the prevalence of diabetic cardiomyopathy.Among the 5 variables,E/A value and Ang? have the largest impact on the formation of diabetic cardiomyopathy and the deterioration of cardiac function.Conclusion 1.Apelin12 and Ang? have significant correlation,mutual promotion and mutual influence,both of which significantly affect the occurrence and development of diabetic cardiomyopathy,and can be used as a high-risk factor for diabetic cardiomyopathy to control cardiac insufficiency.2.Left ventricular diastolic dysfunction first appears in the stage of insulin resistance,which is not dependent on diabetes and diabetic cardiomyopathy,but obviously affects the occurrence and development of diabetic cardiomyopathy and cardiac insufficiency.3.Apelin12 may be used as a new treatment for left ventricular diastolic dysfunction,insulin resistance and heart failure in diabetic cardiomyopathy in the future.Part III: Apelin12,Entresto and both combined to improve myocardial fibrosis in diabetic ratsBackground Diabetic cardiomyopathy(DCM)is one of the most serious complications of diabetes mellitus.Diabetic cardiomyopathy(DCM)usually in myocardial microvascular wall thickening,narrow lumen formed as the main pathological manifestations,extracellular matrix under the effect of pathological formation deposition and prompted myocardial fibrosis,to a certain extent,increased ventricular wall stiffness gradually lead to cardiac systolic and diastolic function decline in performance,will eventually develop into congestive heart failure.According to current epidemiological studies,about 80 percent of diabetics die of heart failure.Some studies have shown that there are significant differences in serum VEGF levels in patients with diabetic cardiomyopathy,and there is a significant correlation between serum VEGF levels and myocardial dilatation in patients with diabetic cardiomyopathy.Apelin is the endogenous ligand of angiotensin II type 1 receptor related protein APJ,which is jointly involved in various pathophysiological mechanisms in vivo.Apelin-12 is one of the most effective subtypes.Entresto(shakobulovalsartan sodium tablet)is a new type of anti-heart failure drug.It can selectively act on AT1 receptor,weaken the role of RAAS,reduce blood pressure,reduce heart load,improve the regulatory function of natriuretic peptide system,and play a role in natriuretic diuresis,kidney function improvement,vasodilation and heart protection.Entresto's clinical research and application were mainly for the treatment of heart failure and hypertension,and there was little research on the improvement of myocardial fibrosis in diabetes.This research through the chain of urea with cephalosporins(STZ)diabetic rats model was established,exogenous Apelin12,observing rats left ventricular mass index(LVMI),Ang?,VEGF,TGF-beta 1 related cytokines and pathological changes,such as exogenous Entresto,observing rats left ventricular mass index(LVMI),Apelin12,Ang? and TGF-beta 1 related cytokines and pathological changes,to explore its possible mechanism.Objective This research through the chain of urea with cephalosporins(STZ)diabetic rats model was established,exogenous Apelin12,observing rats left ventricular mass index(LVMI),Ang?,VEGF,TGF-beta 1 related cytokines and pathological changes,such as exogenous Entresto,observing rats left ventricular mass index(LVMI),Apelin12,Ang? and TGF-beta 1 related cytokines and pathological changes,to explore its possible mechanism.Materials and methods Sixty male rats were randomly divided into two groups: 10 in the control group(NC group)and 50 in the diabetic model group(STZ group).The control group was given a normal diet,and the experimental group was given a high-sugar and high-fat diet.After 4 weeks,STZ(streptozotocin)was injected once into the left lower quadrant of the experimental group at a dose of 55 mg/kg per injection,and the control group was injected with an equal amount of citrate buffer solution.According to the film formation standard of diabetic rats(46 films were formed,the film formation rate was 92%).After successful model making,the diabetic model group was randomly divided into 4 groups,16 in the STZ group,10 in the Apelin12 treatment group,10 in the Entresto treatment group,and 10 in the combined treatment group.The NC group and the STZ group were given daily intervention in the same volume of autoclaved water.The Apelin12 treatment group was daily dissolved with Apelin12 at a dose of 75 ug/kg and autoclaved water.The Entresto treatment group was given Entresto daily.Fully dissolved in autoclaved water,at a dose of 5 mg/kg bid,Apelin 12 + Entresto combination treatment group,daily Apelin 12 at a dose of 75 ug / kg and Entresto at a dose of 5 mg / kg bid,all three groups were continuously administered 12 Week(weigh the weight once a week,adjust the dose according to the weight in time).At week 24,the fasting blood glucose,blood lipids,fasting insulin,HOMA-IR values were monitored;left ventricular mass index(LVMI)was calculated;the thoracic cavity was immediately opened,the heart was quickly removed,and part of the tissue was extracted from the left ventricular myocardium into a 10% formaldehyde solution.Fixing is carried out,and some tissues which have been fixed are dehydrated,and then the dehydrated sample tissue is embedded with paraffin,and sliced according to conventional requirements for HE staining and Masson staining,and partially taking 100 mg of myocardial tissue.After grinding,lysis for 30 minutes,centrifugation,and the supernatant was stored,and the protein levels of Apelin12,Ang?,VEGF and TGF-?1 were determined by enzyme-linked immunosorbent assay(ELISA).Results FPG,FINS,Hb A1 C,HOMA-IR,left ventricular mass index(LVMI),and LVEDd were significantly higher in the STZ group than in the control group,P<0.05.Compared with the STZ group,the Apelin12 treatment group,the Entresto treatment group,FPG,FINS There were differences between Homa-IR,LVMI and LVEDd(P<0.05).The combination of FPG,Homa-IR,LVMI and STZ in the combination group was P<0.01,while the FINS,LVEDd and STZ groups in the combination group were P<0.05.After intervention with Entresto,Apelin12 was significantly higher in diabetic rats than in STZ group(P<0.01),and left ventricular mass index(LVMI)was decreased(P<0.05).HE staining showed that rat myocardial cells in the model group were disordered,with myocardial cells distorted and hypertrophy,inflammatory cells infiltrating and increased fibroblast formation.The myocardial cells of the control group were orderly arranged with clear structure,and only a few fibroblasts were visible.Between the Apelin12 treatment group and Entresto treatment group and the two groups,the improvement was more obvious in the combination treatment group.Masson staining results: the collagen fibers in the myocardial stroma of rats in the model group increased significantly,with disordered arrangement and increased pericapillary matrix.In the control group,only a small amount of collagen fibers existed between the perivascular and myocardial cells.The Apelin12 treatment group and Entresto treatment group had significantly less collagen fibers stained dark green in the observation points and patches than the model group,and the combination treatment group had significantly less collagen fibers than the monotherapy group.After intervention by Apelin12,collagen fibers in myocardial stroma were significantly reduced,local Ang? concentration and protein expression of TGF-beta 1 were decreased(P<0.05),and VEGF concentration was increased(P<0.05).By Entresto intervention,the myocardial interstitial collagen fiber decreased,local Ang? concentration and protein expression of TGF-beta 1 decreased(P<0.05),and Apelin12 concentration increased(P<0.05)in diabetic rats.CVF significantly increased in STZ group compared with that in control group.Exogenous treatment with Apelin12 and Entresto significantly decreased CVF,P<0.05.Conclusion 1.Apelin12 fights against myocardial fibrosis,promotes myocardial angiogenesis,and improves myocardial fibrosis in diabetic rats by affecting Ang? and VEGF metabolic pathways.Entresto inhibited RAS system activation,improved cardiac stress load,increased Apelin12,and significantly improved myocardial fibrosis in diabetic rats.3.Apelin12 and Entresto influenced and coordinated each other in improving myocardial fibrosis in diabetic rats,suggesting that part of the mechanism was crossover.In the future,they may become new drugs for diabetes mellitus combined with cardiomyopathy.