| Dissertation ⅠEffects and mechanisms of Setanaxib(GKT137831)in the treatment of Doxorubicin-induced cardiotoxicityBackgroundWith the continuous development of cancer treatments including chemotherapy,radiotherapy and immunotherapy,the prognosis and survival rate of patients with cancer have been significantly improved.However,among some cancer survivors,cardiovascular disease has become the main cause of death.On the one hand,there are many common risk factors and similarities in pathophysiological mechanisms between cardiovascular disease and cancer,and patients may suffer from both diseases.On the other hand,cardiovascular damage caused by cancer treatment may make these survivors face greater risks of cardiovascular disease.Therefore,cancer treatment and cardiovascular disease have entered the era of convergence,and Cardio-oncology,a new interdisciplinary subject,has emerged as the times require.Cancer treatment-related cardiac dysfunction(CTRCD)is the most worrying cardiovascular complication in cancer treatment,leading to an increase in mortality.Doxorubicin(DOX)is a classic anthracycline antibiotic with high efficiency and broad-spectrum,which can be used to treat solid tumors(such as breast cancer,ovarian cancer and gastrointestinal malignancies)and hematological malignancies(such as lymphoma and leukemia)in adults and children.However,it can cause time and dose-dependent cardiotoxicity,leading to severe cardiomyopathy and congestive heart failure,which limit its clinical application.The mechanism of DOX-induced cardiotoxicity may be multifaceted,including the direct way of increasing the production of reactive oxygen species(ROS)and affecting topoisomerase 2β,and other indirect ways.NADPH oxidase(NOX)is a multi-component enzyme complex,which is the main source of ROS in eukaryotic cells.ROS produced by NOX is involved in numerous crucial physiological processes and is a key regulator of signal transduction pathways.NOX were found to be associated with DOX-induced cardiomyocyte apoptosis.For example,gp91phox knockout or NOX2 knockout mice can resist DOX-induced cardiotoxicity;NOX2 knockout significantly reduced DOX-induced myocardial atrophy,apoptosis,fibrosis,leukocyte infiltration and cardiac dysfunction in mice.The deletion of Rac1 gene in cardiomyocytes and the Rac1 inhibitor NSC23766 also protected DOX-induced cardiotoxicity.Therefore,targeting NOX may be an effective method to reduce DOX-induced cardiotoxicity.Setanaxib(GKT137831)is a specific dual inhibitor of NOX1 and NOX4.Recent studies showed that GKT137831 has a protective effect on the occurrence and development of many cardiovascular diseases.For example,GKT137831 delayed the development of diabetes-related atherosclerosis,and reduced myocardial hypertrophy and remodeling caused by hypertension.However,there are few studies on whether GKT137831 has a protective effect on DOX-induced cardiotoxicity.In this study,we investigated the role of GKT137831 in ameliorating DOX-induced cardiotoxicity and its potential mechanisms.Objectives1.To explore whether GKT137831 can ameliorate DOX-induced cardiotoxicity.2.To explore the potential mechanisms of GKT137831 in ameliorating DOX-induced cardiotoxicity.Methods1.Establishment of DOX-induced cardiotoxicity animal model and interventionSixty 8-week-old male C57BL/6J mice were randomly divided into four groups:control group(n=10),control+GKT137831 group(n=10),DOX group(n=20),and DOX+GKT137831 group(n=20).To establish DOX-induced cardiotoxicity animal model,DOX group and DOX+GKT137831 group mice were administered intraperitoneally DOX(5mg/kg)with a cumulative dose of 20mg/kg.After the first dose of DOX,mice from the control+GKT137831 group and DOX+GKT137831 group were treated with GKT137831(60mg/kg/d)by gavage for 6 weeks.2.EchocardiographyAt the end of the experiment,transthoracic echocardiography was performed.The parameters included left ventricular ejection fraction(LVEF),short axis shortening rate(FS%),end systolic ventricular septal thickness(IVSS),end diastolic ventricular septal thickness(IVSD),left ventricular end systolic diameter(LVESD),left ventricular end diastolic diameter(LVEDD),left ventricular end systolic posterior wall thickness(LVPWS)and left ventricular end diastolic posterior wall thickness(LVPWD)were collected.Five consecutive cardiac cycles were selected for measurement,and the average of the five measurements was taken.3.Collection of mouse tissuesAfter 6 weeks gavage,mice were euthanized,and cardiac tissue and blood samples were collected.Blood samples were centrifuged and frozen at-80℃.For heart tissues,some tissues were frozen by liquid nitrogen,and then stored in-80℃.Another part of the heart tissue was directly prepared into frozen sections or fixed in 4%paraformaldehyde for subsequent histopathological examination.4.Histopathological examinationThe heart tissue specimen was fixed and paraffin-embedded to prepare paraffin sections.HE staining was performed to observe the morphological changes.Masson staining was used to evaluate fibrosis.The protein expressions of NOX1 and NOX4 were assessed by immunofluorescence staining.The morphological changes of mitochondria were observed by transmission electron microscopy(TEM).TUNEL staining was performed to evaluate apoptosis.DHE staining was used to assess the level of ROS.5.Serological testAt the end of the experiment,the serum samples were collected and the concentrations of N-terminal B type natriuretic peptide precursor(NT-proBNP),troponin Ⅰ(cTnⅠ)and malondialdehyde(MDA)were measured.6.Extraction and treatment of neonatal rat cardiomyocyte(NRCM)NRCM was extracted and seeded in the appropriate cell culture plates.Twenty-four hours after plating,the culture medium was changed to high glucose DMEM supplemented with 1.6%horse serum,1%newborn bovine serum,penicillin(100 U/ml),streptomycin(100mg/ml)and 100μmol/L BRDU.After 48 hours,NRCM was subjected to various treatments as described below.NRCM was treated with different concentrations of DOX for 24 hours or pretreated with GKT137831(5μmol/L)for 1 hour,with NAC(10mmol/l)for 1 hour,with SP600125(10μmol/L)for 1 hour,with PD98059(10μmol/L)for 1 hour,with SB203580(10μmol/L)for 1 hour and then stimulated with DOX(2μmol/L)for 24 hours.7.Detection of lactate dehydrogenase(LDH)The levels of LDH from NRCM were detected with LDH detection kit.8.Detection of reactive oxygen species(ROS)The levels of ROS in NRCM were assessed using the ROS detection kit.9.Detection of mitochondrial membrane potential(ΔΨm)The levels of ΔΨm in NRCM were determined using a ΔΨm detection kit.10.TUNEL assayTUNEL assay kit was used to detect the level of apoptosis in vitro.11.Molecular detectionIn vivo and in vitro experiments,the expression of protein was detected by Western blot.12.Statistical analysisResults1.GKT137831 treatment reduced DOX-induced myocardial injury(1)After six weeks of treatment with GKT137831,Kaplan-Meier survival curve analysis showed that the survival rate of mice in the DOX group was significantly lower than that of the control group(p<0.05).Compared with DOX group,GKT137831 treatment improved the survival rate of mice,but there was no significant difference(p>0.05).(2)After six weeks of GKT137831 treatment,serum NT-proBNP(p<0.001)and cTnⅠ(p<0.001)levels in DOX group were significantly higher than those in control group.Compared with DOX group,GKT137831 treatment significantly reduced the levels of serum NT-proBNP(p<0.01)and cTnⅠ(p<0.01).2.GKT137831 treatment improved DOX-induced cardiac dysfunction(1)Compared with the control group,the LVEF(p<0.001),FS%(p<0.001),IVSS(p<0.01),IVSD(p<0.01),LVPWS(p<0.001)and LVPWD(p<0.01)in the DOX group were significantly decreased,while the LVESD(p<0.001)was significantly increased.The LVEDD was increased,but the difference was not significant(p>0.05).(2)GKT137831 treatment significantly improved DOX-induced left ventricular dilatation,and significantly improved the reduction of LVEF(p<0.01)and FS%(p<0.05).3.GKT137831 treatment improved NRCM viability(1)In vitro,DOX stimulation caused an increase in LDH release in a dose-dependent manner,indicating a decrease in cell viability.(2)The intervention of GKT137831 decreased the release of LDH,suggesting that it could improve NRCM viability4.DOX stimulation increased protein expressions of NOX1 and NOX4 in the myocardium(1)In vivo,compared with the control group,Western blot analysis showed that the protein levels of NOX1 and NOX4 were significantly increased in DOX-treated mouse myocardium(p<0.01).This result was also confirmed by immunofluorescence staining.(2)In vitro,with the increase of DOX stimulation concentration and the prolongation of stimulation time,the protein expressions of NOX1 and NOX4 in NRCM increased in a dose and time-dependent manner.5.GKT137831 treatment reduced the excessive oxidative stress of myocardium induced by DOX(1)In vivo,Western blot analysis showed that protein expression levels of NOX1(p<0.05)and NOX4(p<0.01)in the myocardium of the GKT137831-treated mice were significantly reduced compared with the DOX group.In vitro,GKT137831 treatment significantly reduced the protein expression of NOX1(p<0.05)and NOX4(p<0.05)in NRCM.(2)In vivo,compared with the control group,Western blot-determined level of 4-HNE in myocardium of DOX group was significantly increased(p<0.001),while GKT137831 treatment significantly reduced the protein expression of 4-HNE(p<0.01).Compared with the control group,the level of serum MDA in DOX group was significantly increased(p<0.001),while GKT137831 treatment significantly reduced the concentration of MDA(p<0.01).Compared with the control group,the DHE fluorescence intensity in the DOX group was significantly enhanced(p<0.001),while GKT1 37831 treatment significantly reduced DHE fluorescence intensity(p<0.01).(3)In vitro,DCFH-DA indicated that GKT137831 pretreatment significantly reduced DOX-induced ROS production(p<0.05).6.GKT137831 treatment improved DOX-induced myocardial mitochondrial injury(1)In vivo,TEM showed that DOX induced abnormal changes in mitochondrial structure,including irregular arrangement,swelling,vacuolated and disrupted cristae in the mouse heart.The effects were alleviated by GKT137831 treatment.(2)In vitro,JC-1 staining indicated that ΔΨm disruption triggered by DOX was partially restored by intervention of GKT137831.7.GKT137831 treatment attenuated DOX-induced cardiomyocyte apoptosis(1)In vivo,compared with the control group,the proportion of TUNEL positive cells was significantly increased in the DOX-treated mouse myocardium(p<0.001).In contrast,fewer TUNEL-positive cells were observed in the GKT137831-treated mice subjected to DOX(p<0.001).(2)Western Blot results showed that compared with the control group,the expression levels of apoptosis indicators cleaved PARP(c-PARP)(p<0.001),Bax(p<0.001)and cleaved caspase3(CC3)(p<0.01)in mice of DOX group were significantly increased.The expression level of anti-apoptotic index Bcl-2 was significantly reduced(p<0.001).After treatment with GKT137831,the expression levels of c-PARP(p<0.05),Bax(p<0.01)and CC3(p<0.05)were significantly reduced,while the expression level of Bcl-2 was significantly increased(p<0.01).8.GKT137831 treatment attenuated DOX-induced apoptosis in NRCMs(1)In vitro,compared with the control group,the expression levels of apoptotic indicators c-PARP(p<0.001),Bax(p<0.001)and CC3(p<0.01)in the DOX group were significantly increased,while the expression level of anti-apoptotic indicator Bcl-2 was significantly decreased(p<0.001).After pretreament with GKT137831,the expression levels of c-PARP(p<0.05),Bax(p<0.01)and CC3(p<0.05)were significantly reduced,and the expression level of Bcl-2 was significantly increased(p<0.05).(2)TUNEL staining revealed that pretreament with GKT137831 significantly reduced the percentage of TUNEL-positive cells(p<0.001).9.GKT137831 ameliorated DOX-induced apoptosis by inhibiting MAPK pathway(1)DOX enhanced the phosphorylation and activation of JNK,ERK and p38 MAPK in a time-dependent manner.(2)After pretreatment with SP600125(JNK inhibitor),PD98059(ERK inhibitor)and SB203580(p38 MAPK inhibitor)for 1 hour,NRCMs were incubated with DOX for 24 hours.Western blot analysis demonstrated that the expression levels of apoptosis indicators c-PARP and CC3 were significantly decreased.TUNEL staining also showed that pretreatment with SP600125,PD98059,and SB203580 significantly reduced the percentage of TUNEL-positive cells.(3)NRCMs were pretreated with GKT137831 for 1 hour,and then incubated with DOX for 24 hours.Western blot results showed that the phosphorylation levels of JNK,ERK and p38 MAPK were decreased in DOX-treated NRCMs.Conclusions1.GKT137831 treatment relieved DOX-induced myocardial injury and improved the cardiac function.2.GKT137831 treatment inhibits the excessive oxidative stress of myocardium induced by DOX,ameliorated DOX-induced myocardial mitochondrial damage and apoptosis.3.GKT137831 reduced DOX-induced apoptosis by inhibiting MAPK pathway.Effects and mechanisms of Empagliflozin in the treatment of heart failure with preserved ejection fractionBackgroundThe incidence of heart failure with preserved ejection fraction(HFpEF)accounts for more than 50%of heart failure(HF).With the aging of the population and the increasing incidence of metabolic diseases,such as hypertension,diabetes and obesity,the incidence and prevalence of HFpEF are also on the rise.In addition to the heavy burden of disease and poor prognosis,HFpEF patients have a high re-hospitalization rate and a poor quality of life,with a mortality rate of 10%-30%in 5 years.Therefore,HFpEF has become a serious public health problem.So far most drugs approved for the treatment of heart failure with reduced ejection fraction(HFrEF)fail to reduce mortality rates in HFpEF,which is largely due to the complexity and heterogeneity of the disease itself.Therefore,in order to improve the understanding of the pathogenesis and treatment of HFpEF,and better study the effects and mechanisms of inflammation,macrovascular and microvascular dysfunction,fibrosis and tissue remodeling in the progression of HFpEF,it is particularly important to construct a good animal model of HFpEF.At the same time,the lack of animal models that can fully reflect the complex situation of human HFpEF has limited the development of therapeutic methods for HFpEF.Empagliflozin(EMPA)is a sodium glucose co-transporter 2 inhibitor(SGLT2i)that reduces glucose re-absorption and blood glucose levels by inhibiting sodium glucose co-transporter 2(SGLT2)in the proximal renal tubule S1.More and more clinical trials have confirmed that SGLT2i can significantly reduce the hospitalization rate of patients due to HF,and its clinical benefit is not related to whether the patients are accompanied with diabetes or not.Even with HFpEF,which so far does not have a particularly effective therapies,SGLT2i shows some encouraging results.As SGLT2i,EMPA becoming the first drug that can significantly improve the clinical outcomes of HFpEF,while Dapagliflozin(DAPA)can reduce the risk of cardiovascular death,all-cause death and major adverse cardiovascular events(MACE)on the total ejection fraction spectrum.All these remind us that the specific mechanism of SGLT2i clinical benefit in HF deserves more basic research,especially in the field of HFpEF.In this study,we explored the effects and potential mechanisms of EMPA in the treatment of HFpEF on the basis of a mouse model with HFpEF intervened by composite factors,in order to provide new ideas for the treatment of HFpEF.ObjectivesTo investigate the role and mechanism of empagliflozin in the treatment of heart failure with preserved ejection fraction.Methods1.Establishment of HFpEF animal model and interventionSixty 8-week-old male C57BL/6N mice were adaptively fed for one week and then randomly divided into two groups:the control group(n=20)and the HFpEF model group(n=40).The HFpEF model was established using a high-fat diet(HFD)+L-NAME(0.5g/L in drinking water).After co-feeding with HFD+L-NAME for five weeks,the two groups of mice were randomly divided into four groups:control group(n=10),EMPA group(n=10),HFpEF model group(n=20),and HFpEF+EMPA group(n=20).Mice in the HFpEF+ EMPA group were treated with EMPA(10mg/kg/day)by gavage for 6 weeks with continuous HFD+L-NAME co-feeding,while mice in the EMPA group were also given EMPA(10mg/kg/day)by gavage for 6 weeks.Meanwhile,the mice in the control group and the HFpEF model group were orally administered with an equal volume of 0.5%hydroxyethyl cellulose(0.5%hydroxyethyl cellulose as the solvent of EMPA)for 6 weeks.The mice were euthanized after 6 weeks gavage.2.Assessment of general conditions of miceAt the end of the experiment,body weight,blood pressure,glucose tolerance,lung weight and exercise tolerance of mice in different groups were measured and evaluated.3.EchocardiographyAt the end of the experiment,transthoracic echocardiography was performed.Parameters included left ventricular ejection fraction(LVEF),fractional shortening(FS%),left ventricular end systolic diameter(LVESD),left ventricular end diastolic diameter(LVEDD),end systolic ventricular septal thickness(IVSS),end diastolic ventricular septal thickness(IVSD),left ventricular end systolic posterior wall thickness(LVPWS),left ventricular end diastolic posterior wall thickness(LVPWD),peak mitral flow velocity in early diastole(E),peak mitral flow velocity in late diastole(A),and peak mitral annular diastolic velocity tissue doppler(E’)were collected.Five consecutive cardiac cycles were selected for the above measurements,and the average of the five measurements was taken.4.Collection of mouse tissuesAfter 6 weeks gavage,the mice were euthanized,and samples were taken.Blood samples were centrifuged and frozen at-80℃.For heart tissues,some tissues were frozen by liquid nitrogen,and then stored in-80℃.Another part of the heart tissue was directly prepared into frozen sections or fixed in 4%paraformaldehyde for subsequent histopathological examination.5.Histopathological examinationThe heart tissue specimen was fixed and paraffin-embedded to prepare paraffin sections.HE staining was performed to observe the morphological changes.Masson staining was used to observe the degree of fibrosis.WGA staining was performed to calculate the myocardial cell cross-sectional area.Immunohistochemical staining was performed to observe the contents of TNF-α and Titin.DHE staining was performed to assess the condition of the ROS.6.Serological testAt the end of the experiment,the serum samples were collected and the concentrations of inflammatory factors,malondialdehyde(MDA)and cyclic guanosine monophosphate(cGMP)were measured.7.Total nitric oxide(NO)detectionA proper amount of myocardial tissue was prepared into homogenate and the level of NO in the myocardium was determined with total nitric oxide(NO)detection kit.8.ATP detectionA proper amount of myocardial tissue was prepared into homogenate and the supernatant was taken,followed by determination of ATP content in myocardium with ATP detection kit.9.Molecular detectionIn vivo experiments,the expression of protein was detected by Western blot.10.Statistical analysisResults1.EMPA treatment reduced body weight,blood pressure,and improved glucose tolerance and exercise tolerance in HFpEF mice(1)After six weeks of EMPA treatment,the body weight of HFpEF mice was significantly reduced as compared with those in the HFpEF group(p<0.001).(2)After six weeks of EMPA treatment,the systolic blood pressure(p<0.001)and diastolic blood pressure(p<0.01)of the HFpEF mice were significantly reduced as compared with those in the HFpEF group.(3)Compared with the HFpEF group,EMPA treatment significantly improved the glucose tolerance of HFpEF mice(p<0.01).(4)After six weeks of EMPA treatment,the lung weights(wet lung weight/dry lung weight)of HFpEF mice were decreased without significant difference as compared with those in the HFpEF group(p>0.05).(5)Compared with the HFpEF group,after six weeks of EMPA treatment,the exercise tolerance of HFpEF mice was significantly improved,both the running distance(p<0.001)and running time(p<0.001)were significantly increased.2.EMPA treatment reduced myocardial hypertrophy in HFpEF mice(1)Compared with the control group,the myocardium of HFpEF group exhibited concentric hypertrophy.WGA staining showed a significant increase in the cross-sectional area of myocardial cells(p<0.001),the heart weight/tibial length(HW/TL)ratio(p<0.001),and qPCR results showed significantly increased expression of ANP(p<0.001),BNP(p<0.001),and β-MHC(p<0.001)genes related to myocardial hypertrophy.(2)After EMPA treatment,the cross-sectional area of myocardial cells in HFpEF mice was significantly reduced(p<0.001),the HW/TL ratio was significantly reduced(p<0.001),and the expressions of ANP(p<0.05),BNP(p<0.05),and β-MHC(p<0.01)were significantly reduced.3.EMPA treatment improved diastolic function in HFpEF mice(1)With respect to contraction function,there were no significant differences between groups in LVEF(p>0.05)and FS%(p>0.05).(2)Compared with the control group,IVSS(p<0.001),IVSD(p<0.001),LVPWS(p<0.01)and LVPWD(p<0.001)in HFpEF group were significantly increased.LVEDD and LVESD decreased,but there was no significant difference(p>0.05).LVPWS(p<0.05)and LVPWD(p<0.01)were significantly decreased in the HFpEF+EMPA group after treatment with EMPA,but LVESD(p>0.05),LVEDD(p>0.05),IVSS(p>0.05)and IVSD(p>0.05)did not improve significantly.(3)Compared with the control group,the diastolic function in HFpEF mice was damaged,manifested as a significant increase in E/E’(p<0.001).E/A tended to increase,but there was no significant difference(p>0.05).After treatment with EMPA,cardiac diastolic function of HFpEF mice was improved,and E/E’ was significantly decreased(p<0.05).4.EMPA treatment reduces inflammation levels in HFpEF mice(1)Compared with the control group,the levels of serum inflammatory cytokines in HFpEF mice were significantly increased,and EMPA treatment significantly reduced the levels of proinflammatory cytokines.(2)Western blot and immunohistochemical analyses showed that compared with the control group,the protein expressions of IL-1β(p<0.01),IL-6(p<0.001)and TNF-α(p<0.001)in the myocardium were significantly increased in the HFpEF group.However,the protein expressions of IL-1β(p<0.05),IL-6(p<0.05)and TNF-α(p<0.001)in the myocardium of HFPEF mice were significantly decreased after treatment with EMPA.5.EMPA treatment reduced myocardial oxidative stress in HFpEF mice(1)Compared with the control group,the DHE fluorescence intensity in the myocardium of HFpEF group was significantly enhanced(p<0.001),and the DHE fluorescence intensity in the myocardium of EMPA-treated mice was significantly decreased(p<0.001).(2)Compared with the control group,the protein expression of 4-HNE was significantly increased in the myocardium of HFpEF group(p<0.01),and the serum MDA concentration of HFpEF group was also significantly increased(p<0.01).However,EMPA treatment significantly reduced the levels of 4-HNE(p<0.05)and MDA(p<0.05).6.EMPA treatment increased the cardiac NO bioavailability and improved the NO-cGMP-PKG pathway in HFpEF mice(1)The expression of eNOS monomer in the myocardium of HFpEF mice was significantly increased(p<0.001),as well as the eNOS monomer activated by Ser1177 phosphorylation(p<0.001).However,EMPA treatment significantly decreased the protein expression and phosphorylation of eNOS monomer(p<0.05 and p<0.001).(2)NO-cGMP-PKG pathway was inhibited in the HFpEF mouse model,which was manifested as significant reduction of NO metabolites in myocardial homogenate(p<0.01),serum cGMP content(p<0.001),and PKG protein expression in myocardium(p<0.01).The immunohistochemistry results showed that the total protein expression of Titin was significant increased(p<0.001).After treatment with EMPA,the NO metabolites in the myocardial homogenate and serum cGMP content were significantly increased(p<0.01),and the PKG protein expression in the myocardium was significantly increased(p<0.01).The immunohistochemistry results showed that the total protein expression of Titin was significantly decreased(p<0.05),suggesting that the phosphorylation of Titin was increased.7.EMPA treatment improved myocardial energy metabolism and increased ATP production in HFpEF mice.(1)In lipid metabolism,the expression of proteins involved in fatty acid oxidation significantly increased in HFpEF mice(p<0.05),including Cptlb,Cpt2,Cd36,Acadm,Acad10,Hadh,Acot2 and Decr1.EMPA treatment did not change the expression level of key proteins in fatty acid oxidation.The expression of adiponectin(Adipoq)in HFpEF mice was decreased(p<0.05),but significantly increased after EMPA treatment(p<0.05).(2)In carbohydrate metabolism,after EMPA treatment,the phosphorylation level of PDH,a key enzyme involved in glucose aerobic oxidation,was significantly decreased(p<0.05).Among the enzymes involved in glycolytic pathway,Hk2 and Eno3 were significantly decreased(p<0.05).The expression of Pfkl and Pkm were decreased after EMPA treatment,but there was no significant difference(p>0.05).(3)In ketone metabolism and branched-chain amino acid(BCAAs)metabolism,the expression of Bdhl,the key enzyme of ketone body catabolism,was significantly decreased in HFpEF mice(p<0.05),but its expression level was not affected by EMPA treatment.The expression of Bckdha,the rate-limiting enzyme of BCAAs catabolism,was remarkably decreased in the heart of HFpEF mice(p<0.05),but its expression level was not affected by EMPA treatment.(4)In the myocardium of HFpEF mice,the expressions of mitochondrial complex Ⅰ-Ⅳrelated components,such as Ndufs4,Sdha,Uqcrfs1 and Ndufa4,were significantly decreased(p<0.05),while EMPA treatment significantly improved the expression of mitochondrial complex Ⅰ-Ⅳ related components.After EMPA treatment,the expression levels of ATP synthase,such as Atp5fld and Atpaf1,were significantly increased,which indicated that EMPA treatment increased the production of ATP in myocardium.Conclusions1.EMPA treatment improved the glucose tolerance and exercise tolerance of HFpEF mice.2.EMPA treatment reduced myocardial hypertrophy in HFpEF mice.3.Empa treatment improved the diastolic function of HFpEF mice through NO-cGMP-PKG pathway.4.EMPA treatment improved the myocardial energy metabolism of HFpEF mice. |
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