Jianpi Qinghua Formula Alleviates Diabetic Myocardial Injury Through Inhibiting JunB/c-Fos Expression

Objective Diabetic cardiomyopathy(DCM)represents a substantial risk factor for heart failure and increased mortality in individuals afflicted with diabetes mellitus(DM).DCM typically manifests as myocardial fibrosis,myocardial hypertrophy,and impaired left ventricular diastolic function.While the clinical utility of the Jianpi Qinghua(JPQH)formula has been established in treating diabetes and insulin resistance,its potential efficacy in alleviating diabetic cardiomyopathy remains uncertain.This study aims to investigate the impact and underlying molecular mechanisms of the JPQH formula(JPQHF)in ameliorating myocardial injury in nonobese diabetic rats,specifically focusing on apoptosis and inflammation.Methods Wistar rats were assigned as the normal control group(CON),while Goto-Kakizaki(GK)rats were randomly divided into three groups:DM,DM treated with the JPQHF,and DM treated with metformin(MET).Following a 4-week treatment regimen,various biochemical markers related to glucose metabolism,cardiac function,cardiac morphology,and myocardial ultrastructure in GK rats were assessed.RNA sequencing was utilized to analyze differential gene expression and identify potential therapeutic targets.In vitro experiments involved high glucose to induce apoptosis and inflammation in H9c2 cells.Cell viability was evaluated using CCK-8 assay,apoptosis was monitored via flow cytometry,and the production of inflammatory cytokines was measured using quantitative real-time PCR(qPCR)and ELISA.Protein expression levels were determined by Western blotting analysis.The investigation also incorporated the use of MAPK inhibitors to further elucidate the mechanism at both the transcriptional and protein levels.Results The JPQHF group exhibited significant reductions in interventricular septal thickness at end-systole(IVSs)and left ventricular internal diameter at end-systole and end-diastole(LVIDs and LVIDd).JPQHF effectively suppressed high glucose-induced activation of IL-1βand caspase 3 in cardiomyocytes.Furthermore,JPQHF downregulated the expression of myocardial JunB/c-Fos,which was upregulated in both diabetic rats and high glucose-treated H9c2 cells.Conclusion The JPQH formula holds promise in mitigating diabetic myocardial apoptosis and inflammation in cardiomyocytes by inhibiting JunB/c-Fos expression through suppressing the MAPK(p38 and ERK1/2)pathway.

Teneligliptin mitigates diabetic cardiomyopathy by inhibiting activation of the NLRP3 inflammasome

BACKGROUND Diabetic cardiomyopathy(DCM),which is a complication of diabetes,poses a great threat to public health.Recent studies have confirmed the role of NLRP3(NOD-like receptor protein 3)activation in DCM development through the inflammatory response.Teneligliptin is an oral hypoglycemic dipeptidyl peptidase-IV inhibitor used to treat diabetes.Teneligliptin has recently been reported to have anti-inflammatory and protective effects on myocardial cells.AIM To examine the therapeutic effects of teneligliptin on DCM in diabetic mice.METHODS Streptozotocin was administered to induce diabetes in mice,followed by treatment with 30 mg/kg teneligliptin.RESULTS Marked increases in cardiomyocyte area and cardiac hypertrophy indicator heart weight/tibia length reductions in fractional shortening,ejection fraction,and heart rate;increases in creatine kinase-MB(CK-MB),aspartate transaminase(AST),and lactate dehydrogenase(LDH)levels;and upregulated NADPH oxidase 4 were observed in diabetic mice,all of which were significantly reversed by teneligliptin.Moreover,NLRP3 inflammasome activation and increased release of interleukin-1βin diabetic mice were inhibited by teneligliptin.Primary mouse cardiomyocytes were treated with high glucose(30 mmol/L)with or without teneligliptin(2.5 or 5μM)for 24 h.NLRP3 inflammasome activation.Increases in CKMB,AST,and LDH levels in glucose-stimulated cardiomyocytes were markedly inhibited by teneligliptin,and AMP(p-adenosine 5‘-monophosphate)-p-AMPK(activated protein kinase)levels were increased.Furthermore,the beneficial effects of teneligliptin on hyperglycaemia-induced cardiomyocytes were abolished by the AMPK signaling inhibitor compound C.CONCLUSION Overall,teneligliptin mitigated DCM by mitigating activation of the NLRP3 inflammasome.

Empagliflozin ameliorates diabetic cardiomyopathy probably via activating AMPK/PGC-1αand inhibiting the RhoA/ROCK pathway

BACKGROUND Diabetic cardiomyopathy(DCM)increases the risk of hospitalization for heart failure(HF)and mortality in patients with diabetes mellitus.However,no specific therapy to delay the progression of DCM has been identified.Mitochondrial dysfunction,oxidative stress,inflammation,and calcium handling imbalance play a crucial role in the pathological processes of DCM,ultimately leading to cardiomyocyte apoptosis and cardiac dysfunctions.Empagliflozin,a novel glucoselowering agent,has been confirmed to reduce the risk of hospitalization for HF in diabetic patients.Nevertheless,the molecular mechanisms by which this agent provides cardioprotection remain unclear.AIM To investigate the effects of empagliflozin on high glucose(HG)-induced oxidative stress and cardiomyocyte apoptosis and the underlying molecular mechanism.METHODS Twelve-week-old db/db mice and primary cardiomyocytes from neonatal rats stimulated with HG(30 mmol/L)were separately employed as in vivo and in vitro models.Echocardiography was used to evaluate cardiac function.Flow cytometry and TdT-mediated dUTP-biotin nick end labeling staining were used to assess apoptosis in myocardial cells.Mitochondrial function was assessed by cellular ATP levels and changes in mitochondrial membrane potential.Furthermore,intracellular reactive oxygen species production and superoxide dismutase activity were analyzed.Real-time quantitative PCR was used to analyze Bax and Bcl-2 mRNA expression.Western blot analysis was used to measure the phosphorylation of AMP-activated protein kinase(AMPK)and myosin phosphatase target subunit 1(MYPT1),as well as the peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)and active caspase-3 protein levels.RESULTSIn the in vivo experiment, db/db mice developed DCM. However, the treatment of db/db mice with empagliflozin(10 mg/kg/d) for 8 wk substantially enhanced cardiac function and significantly reduced myocardial apoptosis,accompanied by an increase in the phosphorylation of AMPK and PGC-1α protein levels, as well as a decrease inthe phosphorylation of MYPT1 in the heart. In the in vitro experiment, the findings indicate that treatment ofcardiomyocytes with empagliflozin (10 μM) or fasudil (FA) (a ROCK inhibitor, 100 μM) or overexpression of PGC-1α significantly attenuated HG-induced mitochondrial injury, oxidative stress, and cardiomyocyte apoptosis.However, the above effects were partly reversed by the addition of compound C (CC). In cells exposed to HG,empagliflozin treatment increased the protein levels of p-AMPK and PGC-1α protein while decreasing phosphorylatedMYPT1 levels, and these changes were mitigated by the addition of CC. Adding FA and overexpressingPGC-1α in cells exposed to HG substantially increased PGC-1α protein levels. In addition, no sodium-glucosecotransporter (SGLT)2 protein expression was detected in cardiomyocytes.CONCLUSION Empagliflozin partially achieves anti-oxidative stress and anti-apoptotic effects on cardiomyocytes under HGconditions by activating AMPK/PGC-1α and suppressing of the RhoA/ROCK pathway independent of SGLT2.

Integrated mass spectrometry imaging reveals spatial-metabolic alteration in diabetic cardiomyopathy and the intervention effects of ferulic acid

Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity of mass spectrometry(MS)with the spatial information of imaging.In this study,we used MSI to visualize metabolites in the rat heart with high spatial resolution and sensitivity.We optimized the air flow-assisted desorption electrospray ionization(AFADESI)-MSI platform to detect a wide range of metabolites,and then used matrix-assisted laser desorption ionization(MALDI)-MSI for increasing metabolic coverage and improving localization resolution.AFADESI-MSI detected 214 and 149 metabolites in positive and negative analyses of rat heart sections,respectively,while MALDI-MSI detected 61 metabolites in negative analysis.Our study revealed the heterogenous metabolic profile of the heart in a DCM model,with over 105 region-specific changes in the levels of a wide range of metabolite classes,including carbohydrates,amino acids,nucleotides,and their derivatives,fatty acids,glycerol phospholipids,carnitines,and metal ions.The repeated oral administration of ferulic acid during 20 weeks significantly improved most of the metabolic disorders in the DCM model.Our findings provide novel insights into the molecular mechanisms underlying DCM and the potential of ferulic acid as a therapeutic agent for treating this condition.

Hyperglycemia-induced myocardial fibrosis may be associated with pyroptosis and apoptosis of cardiomyoctes in diabetic mice

Myocardial fibrosis is an important manifestation of diabetic cardiomyopathy.This study investigated the potential mechanism of diabetic myocardial fibrosis.Male C57BL/6J and db/db mice aged 8 weeks were randomly divided into the diabetic(DB)and control groups.At 20 weeks,the mouse heart was harvested and subjected to hematoxylin-eosin staining(HE)and Masson staining to investigate the degree of fibrosis.The expressions of transforming growth factor-beta 1(TGF-β1),collagen-III,B-cell lymphoma-2(Bcl2),Bcl2-associated X protein(Bax),cleaved gasdermin D(GSDMD),cysteinyl aspartate specific proteinase-1(caspase-1),apoptosis-associated speck-like protein containing a CARD(ASC),and nucleotide-binding oligomerization domain(NOD)-like receptor 3(NLRP3)were measured by western blotting.Immunohistochemistry and TdT-mediated dUTP nick end labeling(TUNEL)staining were performed to analyze the development of apoptosis and pyroptosis.A significant increase in body weight and blood glucose in the DB group was observed.Myocardial pathological injury,fibrosis,apoptosis,and pyroptosis were more obvious and serious in the DB group.The expression of anti-apoptotic Bcl2 significantly decreased,while the expression levels of pro-apoptotic Bax,caspase-3,and pyroptosis-related proteins,such as cleaved GSDMD,and caspase-1 in the DB group were significantly increased.Pyroptosis and apoptosis were probably the main mechanisms that caused myocardial fibrosis in mice with diabetes.

Molecular targets and mechanisms of Jiawei Jiaotai Pill on diabetic cardiomyopathy based on network pharmacology

BACKGROUND Jiawei Jiaotai Pill is commonly used in clinical practice to reduce apoptosis,increase insulin secretion,and improve blood glucose tolerance.However,its mechanism of action in the treatment of diabetic cardiomyopathy(DCM)remains unclear,hindering research efforts aimed at developing drugs specifically for the treatment of DCM.AIM To explore the pharmacodynamic basis and molecular mechanism of Jiawei Jiaotai Pill in DCM treatment.METHODS We explored various databases and software,including the Traditional Chinese Medicine Systems Pharmacology Database,Uniport,PubChem,GenCards,String,and Cytoscape,to identify the active components and targets of Jiawei Jiaotai Pill,and the disease targets in DCM.Protein-protein interaction network,gene ontology,and Kyoto Encyclopedia of Genes and Genomes analyses were used to determine the mechanism of action of Jiawei Jiaotai Pill in treating DCM.Molecular docking of key active components and core targets was verified using AutoDock software.RESULTS Total 42 active ingredients and 142 potential targets of Jiawei Jiaotai Pill were identified.There were 100 common targets between the DCM and Jiawei Jiaotai Pills.Through this screening process,TNF,IL6,TP53,EGFR,INS,and other important targets were identified.These targets are mainly involved in the positive regulation of the mitogen-activated protein kinase(MAPK)MAPK cascade,response to xenobiotic stimuli,response to hypoxia,positive regulation of gene expression,positive regulation of cell proliferation,negative regulation of the apoptotic process,and other biological processes.It was mainly enriched in the AGE-RAGE signaling pathway in diabetic complications,DCM,PI3K-Akt,interleukin-17,and MAPK signaling pathways.Molecular docking results showed that Jiawei Jiaotai Pill's active ingredients had good docking activity with DCM's core target.CONCLUSION The active components of Jiawei Jiaotai Pill may play a role in the treatment of DCM by reducing oxidative stress,cardiomyocyte apoptosis and fibrosis,and maintaining metabolic homeostasis.

Potential therapeutic targets for the prevention of diabetic nephropathy:Glycyrrhetinic acid

Uncontrolled hyperglycemia or poorly managed disease increases the propensityfor a number of diabetes-related complications targeting major organs includingthe heart, eyes, and kidney. Although the mechanisms by which diabetes inducescardiovascular diseases include oxidative stress and inflammation, when insulinresistance remains the key to the pathogenesis, as implicated in the two reviews inthis issue. This editorial mainly comments on the potential preventive applicationof glycyrrhetinic acid (or 18β-GA) in relation to diabetic nephropathy. The therapeuticor preventive effects of 18β-GA, as a hydrolytic product of glycy-rrhizicacid that is a component of licorice, have been appreciated in other disorders, buthave received much less attention in relation to diabetic complications. A study inthis issue has identified 18β-GA as a therapeutic for preventing diabeticnephropathy and provides evidence to support efficacy in cultured human renaltubule cells in vitro. Although it represents a pilot study, the observations supporta new therapeutic approach that warrants further ex-ploration.

The protective effect and mechanism of rhein on diabetic cardiomyopathy by regulating Sirt1/PGC-1α pathway

Objective:To explore the protective effects of rhein on cardiomyocyte injury in DCM and its possible mecha-nism.Methods:The diabetic model was induced by intraperitoneal injection with streptozotocin and high-fat diet.The mice were randomly divided into control group,DM group,and DM+RH group.After 12 weeks treatment with rhein,the change of fast blood glucose,body weight,and heart weight/body weigh(t HW/BW)were observed.HE and Masson staining were used to evalu-ate myocardial structural damage.Transmission electron microscope was used to observe the myocardial mitochondrial structure.The mRNA levels of Sirt1,PGC-1α,TFAM,ANP,BNP andβ-MHC were quantified by RT-PCR.Sirt1,PGC-1α and TFAM protein levels were estimated by Western blot and IHC.Results:Compared with control group,the blood glucose,HW/BW,ANP,BNP andβ-MHC mRNA of DM group were significantly increased(P<0.05).The structures of myocardium and mitochondria were obviously destroyed in DM group.Sirt1,PGC-1α and TFAM expression were significantly decreased(P<0.05).Compared with DM group,the blood glucose,HW/BW,ANP,BNP and β-MHC mRNA of DM+RH group were decreased(P<0.05).The myocardial and mitochondrial injury were improved.Sirt1,PGC-1α and TFAM expression were significantly increased(P<0.05).Conclusion:Rhein exhibits protective effects on diabetic cardiomyopathy which may be achieved by activating Sirt1/PGC-1α pathway.

Diabetic heart disease:A clinical update

Diabetes mellitus(DM) significantly increases the risk of heart disease,and DMrelated healthcare expenditure is predominantly for the management of cardiovascular complications.Diabetic heart disease is a conglomeration of coronary artery disease(CAD),cardiac autonomic neuropathy(CAN),and diabetic cardiomyopathy(DCM).The Framingham study clearly showed a 2 to 4-fold excess risk of CAD in patients with DM.Pathogenic mechanisms,clinical presentation,and management options for DM-associated CAD are somewhat different from CAD among nondiabetics.Higher prevalence at a lower age and more aggressive disease in DM-associated CAD make diabetic individuals more vulnerable to premature death.Although common among diabetic individuals,CAN and DCM are often under-recognised and undiagnosed cardiac complications.Structural and functional alterations in the myocardial innervation related to uncontrolled diabetes result in damage to cardiac autonomic nerves,causing CAN.Similarly,damage to the cardiomyocytes from complex pathophysiological processes of uncontrolled DM results in DCM,a form of cardiomyopathy diagnosed in the absence of other causes for structural heart disease.Though optimal management of DM from early stages of the disease can reduce the risk of diabetic heart disease,it is often impractical in the real world due to many reasons.Therefore,it is imperative for every clinician involved in diabetes care to have a good understanding of the pathophysiology,clinical picture,diagnostic methods,and management of diabetes-related cardiac illness,to reduce morbidity and mortality among patients.This clinical review is to empower the global scientific fraternity with up-to-date knowledge on diabetic heart disease.

Role of novel biomarkers in diabetic cardiomyopathy

Diabetic cardiomyopathy(DCM)is commonly defined as cardiomyopathy in patients with diabetes mellitus in the absence of coronary artery disease and hypertension.As DCM is now recognized as a cause of substantial morbidity and mortality among patients with diabetes mellitus and clinical diagnosis is still inappropriate,various expert groups struggled to identify a suitable biomarker that will help in the recognition and management of DCM,with little success so far.Hence,we thought it important to address the role of biomarkers that have shown potential in either human or animal studies and which could eventually result in mitigating the poor outcomes of DCM.Among the array of biomarkers we thoroughly analyzed,long noncoding ribonucleic acids,soluble form of suppression of tumorigenicity 2 and galectin-3 seem to be most beneficial for DCM detection,as their plasma/serum levels accurately correlate with the early stages of DCM.The combination of relatively inexpensive and accurate speckle tracking echocardiography with some of the highlighted biomarkers may be a promising screening method for newly diagnosed diabetes mellitus type 2 patients.The purpose of the screening test would be to direct affected patients to more specific confirmation tests.This perspective is in concordance with current guidelines that accentuate the importance of an interdisciplinary team-based approach.

Inhibition of NF-kB and Wnt/β-catenin/GSK3p Signaling Pathways Ameliorates Cardiomyocyte Hypertrophy and Fibrosis in Streptozotocin (STZ)-induced Type 1 Diabetic Rats

Type 1 diabetes mellitus(T1DM)is associated with an increased risk of diabetic cardiomyopathy(DCM).Nuclear factor kappa B(NF-kB)and Wnt/β-catenin/GSK3p have been demonstrated to play pathogenic roles in diabetes.In this study,we evaluated the roles of these two pathways in T1 DM-induced cardiomyopathy in rats.Streptozotocin(STZ)-induced type 1 diabetic rats were treated with pyrrolidine dithiocarbamate(PDTC)or meisoindigo(Me)to inhibit NF-kB and Wnt/β-catenin/GSK3P respectively for 4 or 8 weeks.As compared with untreated diabetic rats,treatment with either PDTC or Me partly attenuated the myocardial hypertrophy and interstitial fibrosis,improved cardiac function,and exhibited reduction in inflammatory reaction.In addition,we found that inhibiting NF-κB and Wnt/β-catenin/GSK3β pathways could regulate glucose and lipid metabolism.The effects were associated with the decrease of NF-κB activity and the downregulation of some proinflammatory cytokines,including tumor necrosis factor-alpha(TNF-α)and interleukin(IL)-2.Our data suggested that the activities of NF-κB and Wnt/β-catenin/GSK3β pathways were both increased and inhibiting NF-κB and Wnt/β-catenin/GSK3β signaling pathways might improve myocardial injury in T1DM rats.

Astragalus Prevents Diabetic Rats from Developing Cardiomyopathy by Downregulating Angiotensin Ⅱ Type2 Receptors' Expression

This study examined the potential roles of astragalus and angiotensin Ⅱ type 2 receptor (AT2) in rats with streptozotocin (STZ)-induced diabetic cardiomyopathy. Of 52 female 4-week-old Wistar rats treated with high glucose and lipid diet to induce insulin resistance, 7 treated with sodium citrate buffer (pH=4.5) served as controls (con1) and the other 45 were treated by intraperitoneal injection (ip) of STZ to induce type 2 diabetes. After 20 weeks, the maximal velocity decrease of pressure per second in left ventricle within the period of isovolumic relaxation (-dp/dt max) was detected by inserting cannula through right carotid artery. Of the 45 rats, 24 with -dp/dt max ≤700 mmHg/s (1 mmHg=0.133 kPa) developing diabetic cardiomyopathy were grouped as follows: 7 treated with double distilled H 2O (ip) were included in control group 2 (con2); other 8 treated with AT2 agonist (CGP42112A, ip) were included in experimental group1 (exp); 9 treated with astragalus (po) constituted experimental group 2 (exp2). All injections lasted 4 weeks (qd) and the heart weight (HW) was recorded. Cardiomyocyte apoptosis index (CAI), mRNA of AT2 and Bcl-2 as well as AT2 and Bcl-2 protein values in cardiomyocytes were also measured. Our results showed that -dp/dt max in exp1, exp2 and con2 were much lower than those in con1 (P<0.01 ). CAI and AT2 in both mRNA and protein in con1 were lower than those in the other three groups (P<0.01). The three parameters above were higher in exp1 but less in exp2 than those in con2, respectively (P<0.01). The three parameters and HW in exp1 were much higher than those in exp2 (P<0.01). Changes of Bcl-2 were opposite to those of AT2. Our results suggested that high expression of AT2 might accelerate the apoptosis of cardiomyocytes in diabetic rats and play an important role in precipitating diabetic cardiomyopathy and astragalus protects diabetic rats from developing cardiomyopathy by downregulating AT2.

Role of cannabinoids and the endocannabinoid system in modulation of diabetic cardiomyopathy

Diabetic complications,chiefly seen in long-term situations,are persistently deleterious to a large extent,requiring multi-factorial risk reduction strategies beyond glycemic control.Diabetic cardiomyopathy is one of the most common deleterious diabetic complications,being the leading cause of mortality among diabetic patients.The mechanisms of diabetic cardiomyopathy are multi-factorial,involving increased oxidative stress,accumulation of advanced glycation end products(AGEs),activation of various pro-inflammatory and cell death signaling pathways,and changes in the composition of extracellular matrix with enhanced cardiac fibrosis.The novel lipid signaling system,the endocannabinoid system,has been implicated in the pathogenesis of diabetes and its complications through its two main receptors:Cannabinoid receptor type 1 and cannabinoid receptor type 2,alongside other components.However,the role of the endocannabinoid system in diabetic cardiomyopathy has not been fully investigated.This review aims to elucidate the possible mechanisms through which cannabinoids and the endocannabinoid system could interact with the pathogenesis and the development of diabetic cardiomyopathy.These mechanisms include oxidative/nitrative stress,inflammation,accumulation of AGEs,cardiac remodeling,and autophagy.A better understanding of the role of cannabinoids and the endocannabinoid system in diabetic cardiomyopathy may provide novel strategies to manipulate such a serious diabetic complication.

Adriamycin-induced cardiomyopathy can serve as a model for diabetic cardiomyopathy ——a hypothesis

Diabetic cardiomyopathy is one of the life threatening complications of diabetes. A number of animal models are being used for studying diabetic cardiomyopathy. In laboratory animal models, induction of cardiomyopathy happens in two stages: first being the induction of diabetic condition and the second being the induction of cardiomyopathy by prolonging diabetic condition. It takes a longer time to develop diabetes with the limited success rate for development of cardiomyopathy. Adriamycin is an effective anticancer drug limited by its major side-effect cardiomyopathy. A number of features of Adriamycin treatment mimics diabetes. We postulate that Adriamycin-induced cardiomyopathy might be used as a model system to study diabetic cardiomyopathy in rodents since a number of features of both the cardiomyopathies overlap. Left ventricular hypertrophy, systolic and diastolic dysfunction, myofibrillar loss, and fibrosis are hallmarks of both of the cardiomyopathies. At the molecular level, calcium signaling, endoplasmic reticulum stress, advance glycation endproduct activation, mitochondrial dysfunction,inflammation, lipotoxicity and oxidative stress are similar in both the cardiomyopathies.The signature profile of both the cardiomyopathies shares commonalities. In conclusion,we suggest that Adriamycin induced cardiomyopathic animal model can be used for studying diabetic cardiomyopathy and would save time for researchers working on cardiomyopathy developed in rodent using the traditional method.

Heart mitochondrial dysfunction in diabetic rats

Diabetic cardiomyopathy,i.e.the ventricular dysfunction in the absence of hypertension or coronary arterial disease,is a common complication of diabetes mellitus that leads to a heightened risk of heart failure and death among diabetic patients.This contractile dysfunction could be associated to mitochondrial dysfunction,in which mitochondrial biogenesis could emerge as a compensatory mechanism triggered in response to hyperglycemia.It has been proposed that nitric oxide synthase activities with enhanced NO production are involved in this process.Alterations in the contractile response and lusitropic reserve were observed in streptozotocin diabetic rats afterβ-adrenergic stimuli.Additionally,tissue O_(2) consumption was declined.A condition of mitochondrial dysfunction with decreased mitochondrial state 3 O_(2) consumption,respiratory control ratio,mitochondrial respiratory complexes activities and ATP production were present in hearts of diabetic animals.We observed an increase in NO production by heart mitochondria and in cytochrome oxidase activity in heart homogenates.The latter suggests an increase of newly formed mitochondria.Thus,the impairment of mitochondrial function with increased mitochondrial biogenesis may precede the onset of diabetic cardiomyopathy.However,mitochondrial biogenesis does not necessarily imply that the resultant mitochondria are functional,which might explain the changes in cardiac energy metabolism occurring in hearts of diabetic rats.

The relationship between Metrnl and diabetic cardiomyopathy and its related molecular mechanism

Objective:To investigate the changes of serum concentration of Metrnl in diabetic cardiomyopathy mice,and the relationship between Metrnl and Diabetic cardiomyopathy(DCM)and its molecular mechanism.Methods:Fifteen male mice were randomly divided into experimental group(DCM+Metrnl),model group(DCM)and control group.Metrnl concentration was measured with an enzyme-linked immunosorbent assay.The experimental group was treated with Metrnl,and the control group and model group were treated with equal volume solvent.Then the myocardial pathological changes,reactive oxygen species and the expression of PPARs and GLUT4 protein and the expression of CD36 and SOD gene were observed after 7 days of administration of recombinant Metrnl.Results:Serum Metrnl concentrations were elevated in DCM(P>0.05).Metrnl reduced the serum concentrations of total cholesterol(TG,P<0.05),triglyceride(TC,P<0.05)and low density lipoprotein cholesterol(LDL-C,P<0.05),while increased high density lipoprotein cholesterol(HDL-C,P<0.05)in DCM.In addition,Metrnl improved the energy metabolism of DCM,decreased the production of reactive oxygen species(ROS)and up-regulated the protein expressions of PPAR-a,PPAR-β/δ,GLUT4 and the expression of SOD in cardiomyocytes,while CD36 gene expression was down-regulated.Conclusion:Serum Metrnl concentrations were elevated in DCM mouse modles.Metrnl improved lipid metabolism and cardiac function in DCM.Besides,it can reduced myocardial oxidative stress injury through PPAR-β/δ,GLUT4 pathway.

Research progress of miRNA in diabetic cardiomyopathy

Diabetic cardiomyopathy is one of the main causes of death of diabetic patients and seriously endangers human health.MiRNA is a type of endogenous non-coding RNA with a length of 18-25 nucleotides.It can regulate gene expression and plays an important role in the development of diabetic cardiomyopathy.This article will review the pathogenesis of diabetic cardiomyopathy and the important role of miRNA in its pathogenesis in order to provide a reference for future research.

Resveratrol ameliorates diabetic myocardial injury through HSF1-mediated ferroptosis

Objective:To observe the effect of resveratrol on the injury of diabetic cardiomyocytes and its effect on HSF1 mediated iron death.Methods:the diabetic cardiomyopathy model was established by high glucose induced H9c2,and H9c2 was exposed to normal glucose concentration as a control.Then the intervention was performed with the corresponding drugs.The cell proliferation level was detected by CCK8 method,the concentrations of LDH,SOD,MDA and iron ions were detected by kit method,and the expression levels of HSF1,apoptotic proteins(Bax and Bcl-2)and iron death marker proteins(ACSL4,GPX4 and SLC7A11)were detected by Western blot;Results:compared with the blank group,the cell activity,SOD activity,the expression of HSF1,Bcl-2,GPX4 and SLC7A11 protein in the model group decreased significantly,and the LDH activity,MDA content,Bax and ACSL4 protein expression,Bax/Bcl-2 ratio and Fe^(2+)content increased significantly(P<0.01).Compared with the model group,the activity of H9c2 cells,the activity of SOD,the expression of HSF1,Bcl-2,GPX4 and SLC7A11 protein increased significantly,and the activity of LDH,the content of MDA,the expression of Bax and ACSL4 protein,the ratio of Bax/Bcl-2 and the content of Fe^(2+)decreased significantly in the resveratrol group(P<0.01).After the intervention,the activity of H9c2 cells,the activity of SOD,the expression of HSF1,Bcl-2,GPX4 and SLC7A11 protein decreased significantly,and the activity of LDH,the content of MDA,the expression of Bax and ACSL4 protein,the ratio of Bax/Bcl-2 and the content of Fe^(2+)increased significantly in si-hsf1 group(P<0.01);Conclusion:resveratrol can inhibit cell iron death and improve high glucose induced cardiomyocyte injury by up regulating the expression of HSF1.

The mechanism of Astragalus-Prunella vulgaris in the treatment of diabetic cardiomyopathy based on network pharmacology and molecular docking

Objective:To study the main chemical components and mechanism of Astragalus and Prunella vulgaris in the treatment of diabetes cardiomyopathy(DCM)based on network pharmacology and in vitro experiments.Methods:The main active components and prediction targets of Astragalus membranaceus and Prunella vulgaris herbal pairs were obtained by TCM Pharmacology database and analysis platform(TCMSP),and the disease genes were retrieved by genecards,OMIM,PharmGKB and drugbank databases.The disease and drug prediction targets were intersected to screen out common potential therapeutic targets.Cytoscape3.7.2 software was used to construct"drug component disease target"interaction network diagram;The PPI network of protein-protein interaction was constructed by using string database.R software was used to analyze the function enrichment of GO and KEGG for drug disease common targets,and autodock Vina 1.1.2 for molecular docking.Finally,the specific mechanism of Astragalus and Prunella vulgaris medicated serum on high glucose stimulated cardiomyocytes was verified in vitro.H9c2 cardiomyocytes were divided into five groups:normal group:low glucose(5.5 mmol/L)culture group,model group:high glucose(33 mmol/L)culture group,5%serum group:high glucose+5%Astragalus membranaceus Prunella vulgaris herb serum culture group,10%serum group:high glucose+10%Astragalus membranaceus Prunella vulgaris herb serum culture group,15%serum group:Hg high glucose+15%Astragalus membranaceus Prunella vulgaris herb serum culture group.MTT assay was used to detect the cell survival rate,and Western blot was used to detect the effect of Astragalus and Prunella vulgaris medicated serum on the expression of AKT1,p-AKT1,MAPK14 and p-MAPK14 proteins.Results:In this study,31 active components of Astragalus and Prunella vulgaris were screened,involving 157 targets of diabetes cardiomyopathy and 178 related signal pathways.The results of network analysis showed that Astragalus and Prunella vulgaris herbs may play a role in the treatment of DCM by acting on key targets such as AKT1,FOS,MAPK1,MAPK8,MAPK14,Jun and key pathways such as PI3K-AKT.Molecular docking showed that Astragalus membranaceus and Prunella vulgaris medicine had good binding between the active components luteolin,quercetin,pistil isoflavone,kaempferol and key targets such as AKT1,MAPK14,MAPK1,FOS,mapk8 and Jun,and the Vina score of luteolin and AKT1 was the lowest.The results in vitro showed that Astragalus and Prunella vulgaris medicated serum significantly improved the inhibition of H9c2 cardiomyocyte proliferation induced by high glucose,and increased the phosphorylation levels of AKT1 and MAPK14 proteins to play a role in the treatment of DCM.Conclusion:Astragalus and Prunella vulgaris have the characteristics of multi-target and multi-channel in the treatment of DCM.Its mechanism may be related to the regulation of the protein expression of p-AKT1 and p-MAPK14.These findings provide a new idea and basis for further experimental study on the mechanism of Astragalus and Prunella vulgaris in the treatment of diabetes cardiomyopathy.

Study on the mechanism of Astragalus in the treatment of diabetic cardiomyopathy based on network pharmacology and its preliminary verification study

Objective:To explore the potential active ingredients and targets of Astragalus,and also to predict the targets and mechanisms of Astragalus in the treatment of diabetic cardiomyopathy.Based on the predicted results,the key signaling pathways were validated in a diabetic cardiomyopathy model mouse.Methods:Compounds and targets in Astragalus were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.The protein names to corresponding"Gene Symbol ID"was convert by STRING database.We obtained targets of diabetic cardiomyopathy data from DisGeNET datasets.The protein-protein interaction network(PPI network)was established using STRING database.Cytoscape 3.6.0 was used to construct a disease-drug-target gene network map and to screen the 10 closest target genes by Cytohuba plug-in.The overlapping genes were then subjected to gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)-based enrichment analysis.Finally,the key molecules of the MAPK signaling pathway were validated by in vitro experiments.Animal experiments were performed using 21 Kunming mice randomly divided into normal group,model group,and Chinese herbal medicine Astragalus group,with seven mice in each group.The myocardium of mice in each group was stained with HE to compare the pathological morphological changes,and Western Blot was also used to compare the key molecules of MAPK signaling pathway,ERK1 and p-p38.Results:Astragalus contained 20 active ingredients with 188 corresponding targets,220 targets related to diabetic cardiomyopathy and 37 targets acting in conjunction with Astragalus.The common targets were imported into the STRING database to obtain a PPI network graph of overlapping genes,with 37 nodes and 391 edges.The PPI network map was imported into Cytoscape 3.6.0 software,and the most significant top 10 hub genes were obtained using the MCC algorithm in the cytoHubba plugin,namely AKT1,TP53,CASP3,MMP9,EGF,IL-10,CXCL8,IL-1β,VEGFA,PPARG.GO functional enrichment analysis yielded 40 entries for biological process(BP),23 entries for cellular component(CC),22 entries for molecular function(MF)and 94 entries for KEGG pathway enrichment screening,mainly involving PI3K-AKT,MAPK,HIF-1,FOXO,TNP pathway and other inflammation or apoptosis regulatory pathways.Animal experiments showed that Astragalus can improve the inflammatory state of myocardial tissue in mice with diabetic cardiomyopathy,and the expression of ERK1 and p-p38 protein in myocardial tissue of mice in the model group was higher than that in the normal group(P<0.05,P<0.01),and after the intervention with Astragalus,the expression of ERK1 and p-p38 protein was significantly lower than that in the model group,and the difference was statistically significant(P<0.05,P<0.01).Conclusion:Astragalus has multi-target,multi-component and multi-pathway action characteristics in the treatment of diabetic cardiomyopathy,which can exert anti-inflammatory and anti-oxidative stress effects by regulating protein expression of MAPK signaling pathway ERK1,p-p38.