To identify and verify the active ingredients from Astragalus membranaceus on hypertensive cardiac remodeling based on network pharmacology and heart RNA-sequencing data. The monomers of A. membranaceus and their intervention target database were established by using network pharmacology. The genes associated to cardiac remodeling were then screened by analyzing cardiac RNA-sequencing data. An overlap between genes related to cardiac remodeling and targets of ingredients form A. membranaceus was collected to obtain monomers with protective effect on hypertensive cardiac remodeling. Angiotensin Ⅱ(AngⅡ)-induced mouse cardiac remodeling model was used to validate the protective effect of active ingredients from A. membranaceus on hypertensive cardiac remodeling. Finally, a total of 81 monomers and 1 197 targets were enrolled in our database. Mouse RNA-sequencing data showed that 983 genes were significantly up-regulated and 465 genes were down-regulation in myocardial tissues of the cardiac remodeling mice as compared with blank group mice, respectively. Ninety-two genes were found via overlapping between genes related to cardiac remodeling and targets, involving 59 monomers from A. membranaceus. Further research found that vanillic acid(VA) could intervene 27 genes associated with hypertensive cardiac remodeling, ranking top 1. Meanwhile, VA could significantly inhibit AngⅡ-induced increase in ratio of heart weight to body weight and heart weight to tibial length, ANP and BNP mRNA levels in myocardial tissues, myocardial tissue damage, cardiac fibrosis level and cardiac hypertrophy level in vivo. Those results showed that network pharmacology screen-based VA has protective effect on AngⅡ-induced cardiac remodeling.
Angiotensin II ; Animals ; Astragalus propinquus/chemistry* ; Heart ; Hypertension/genetics* ; Mice ; Protective Agents/pharmacology* ; Vanillic Acid/pharmacology* ; Ventricular Remodeling/genetics*

BACKGROUNDHepatocyte growth factor (HGF) inhibits the development of pulmonary artery hypertension (PAH) by reducing pulmonary artery pressure and right ventricle (RV) hypertrophy. However, whether HGF can prevent RV remodeling via inhibiting apoptosis in RV cardiomyocytes and decreasing neurohormonal activation remains unknown.

METHODSThe PAH and subsequent RV remodeling in rats were induced by subcutaneous injection of monocrotaline (MCT). The PAH rats were transfected with adenovirus carrying HGF (Ad-HGF) via intratracheal instillation. Three weeks after transfection, the hemodynamics indexes were measured, serum levels for angiotonin II (ANG II) and brain natriuretic peptide (BNP) were determined by ELISA. Histological analysis was used to assess the RV hypertrophy and fibrosis. The cardiomyocyte apoptosis in RV was assayed by TUNEL staining. The mRNA expression of BNP, angiotensin-converting enzyme (ACE), Bax and Bcl-2 in RV was determined by reverse transcriptase polymerase chain reaction (RT-PCR), the protein expression of transforming growth factor (TGF)-β1 and tumor necrosis factor (TNF)-α in RV was determined by Western blotting.

RESULTSHGF treatment significantly decreased the mean PAH, RV systolic pressure, serum ANG II and BNP levels. HGF treatment also significantly decreased the RV hypertrophy, collagen deposition, and the number of apoptotic cardiomyocytes. Moreover, HGF treatmemt significantly decreased the expression of BNP, ACE, Bax, TGF-β1, and TNF-α, while it significantly increased the expression of Bcl-2.

CONCLUSIONSGene transfer of HGF decreases MCT-induced PAH and improves RV remodeling. This effect is mediated not only by improving the hemodynamics but also by decreasing neurohormonal activation and inhibiting cardiomyocytes apoptosis. HGF gene treatment may be an effective strategy for improving RV remodeling in MCT-induced PAH.

Animals ; Apoptosis ; genetics ; physiology ; Hepatocyte Growth Factor ; genetics ; physiology ; therapeutic use ; Humans ; Hypertension, Pulmonary ; metabolism ; therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling ; genetics ; physiology

OBJECTIVETo investigate the relationship between PPARdelta + 294T/C gene polymorphism and lipid profile, obesity and left ventricular hypertrophy (LVH) in patients with metabolic syndrome (MS).

METHODSThis study was conducted in 300 patients with MS and 174 patients with essential hypertension (EH) and 143 patients with type 2 diabetes mellitus (T2DM). MS was diagnosed according to 1999 WHO criteria. Fasting insulin (FINS), fasting blood glucose (FBG), plasma lipids levels were measured, LVH was examined by Doppler echocardiography. The PPARdelta + 294T/C gene polymorphism were analyzed using polymerase chain reaction and subsequently digested by BSLI restriction endonuclease.

RESULTSThe frequencies of the PPARdelta + 294T/C genotypes were not different among three groups. Compared with T2DM and EH, MS patients had significantly higher body mass index (BMI), plasma total cholesterol, TG and LDL-C levels (P < 0.01 or P < 0.05). LVM, LVMI and incidence rate of LVH were significantly higher in MS and EH patients than that in T2DM (P < 0.01). MS patients with CC genotype had significantly higher total cholesterol and LDL-C levels than those with TT and TC genotypes (total cholesterol in CC genotype: 6.13 +/- 1.86 mmol/L vs in TC genotype: 5.14 +/- 1.10 mmol/L, P < 0.05, and CC genotype: 6.13 +/- 1.86 mmol/L vs TT genotype: 4.99 +/- 1.42 mmol/L, P < 0.01; LDL-C in CC genotype: 3.82 +/- 1.52 mmol/L vs in TC genotype: 3.14 +/- 0.88 mmol/L, P < 0.05, and in CC genotype: 3.82 +/- 1.52 mmol/L vs in TT genotype: 2.90 +/- 0.87 mmol/L, P < 0.01). BMI and LVMI in MS patients with C allele carriers (CC + TC) were significantly higher than that of TT genotype (LVMI in CC + TC: 46 +/- 10 g/m(2.7) vs in TT: 44 +/- 10 g/m(2.7); BMI in CC + TC: 26 +/- 3 kg/m(2) vs in TT: 25 +/- 3 kg/m(2), P < 0.05).

CONCLUSIONSIt is indicated that PPARdelta + 294T/C gene polymorphism in subjects with MS may be involved in the occurrence of obesity and dyslipidemia. MS patients with C allele had a predominant LVH than subjects with TT genotype.

Aged ; Body Mass Index ; Diabetes Mellitus, Type 2 ; genetics ; physiopathology ; Female ; Genotype ; Humans ; Hypertrophy, Left Ventricular ; genetics ; physiopathology ; Lipids ; blood ; Male ; Metabolic Syndrome ; genetics ; physiopathology ; Middle Aged ; Obesity ; genetics ; physiopathology ; PPAR delta ; genetics ; Polymorphism, Single Nucleotide ; Ventricular Remodeling

OBJECTIVETo investigate the relationship between expression of Smad3, Smad7 and ventricular remodeling in rats after myocardial infarction.

METHODSMyocardial infarction was induced by left anterior descending coronary artery ligation in rats (n = 11) and sham-operated rats were used as control (n = 10). The rats were sacrificed 8 weeks later. Heart weight/body weight (HW/BW), mean blood pressure, left ventricular end diastolic pressure (LVEDP), collagen content in the un-infarcted area were examined. The mRNA levels of transforming growth factor (TGF)beta(1), Smad 3, Smad7 were determined by RT-PCR.

RESULTCompared with controls, the level of HW/BW, LVEDP and collagen content were significant increased. The mRNA expression of TGFbeta(1) and Smad3 was significantly increased in areas of myocardial infarction, border of the infarction, interventricular septum and right ventricle. The expression of Smad7 mRNA in these areas was decreased.

CONCLUSIONThese results indicated that TGFbeta(1)-Smads signaling was correlated to the ventricular remodeling after myocardial infarction. Smad3 might promote the process while Smad7 inhibit the process.

Animals ; Male ; Myocardial Infarction ; metabolism ; physiopathology ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta ; metabolism ; Ventricular Remodeling

ADAM Proteins ; genetics ; ADAM17 Protein ; Adrenergic beta-Antagonists ; therapeutic use ; Animals ; Male ; Metoprolol ; therapeutic use ; Myocardial Infarction ; drug therapy ; physiopathology ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; Ventricular Function, Left ; drug effects ; Ventricular Remodeling ; drug effects

OBJECTIVETo detect the regulation of angiogenic genes involved in the processes of collateral development.

METHODSMyocardial infarction (MI) scar was induced by cryoinjury in New Zealand rabbits. Four weeks after MI, 24 hours before cell transplantation, bone marrow was aspirated from the right thigh bone and mononuclear bone marrow cells (BMCs) were isolated by Ficoll density gradient centrifugation. Then the mononuclear BMCs (n = 8) or IMDM culture medium (n = 8) were transplanted into infarction scar and the periphery. Four weeks after mononuclear BMCs transplantation, DNA microarray analysis was performed to detect the regulation of angiogenesis-related genes in infarction scar and the periphery. And the differences of angiogenic genes expression were compared among several important growth factors by Western blot.

RESULTSDNA microarray analysis showed the detail regulation of genes involved in the angiogenic processes. There were 15 genes upregulated over 3 times in the infarction scar. In addition, we also found more genes are involved in the process of angiogenesis in its periphery than in the infarction scar (40 genes vs. 15 genes). Western bolt analysis further demonstrated that mononuclear BMCs transplantation was capable of increasing the levels of VEGF, FGF and Angiopoietin-I expression in the infarction scar and its periphery, compared with the control group, P < 0.05.

CONCLUSIONThese findings indicate that the natural angiogenic processes leading to collateral development are extremely complex, since many kinds of bone marrow-derived growth factors involved in the processes after mononuclear BMCs transplantation into infarction sites.

Animals ; Bone Marrow Transplantation ; Female ; Gene Expression Profiling ; Male ; Monocytes ; metabolism ; Myocardial Infarction ; genetics ; pathology ; therapy ; Neovascularization, Pathologic ; metabolism ; Oligonucleotide Array Sequence Analysis ; Rabbits ; Up-Regulation ; Ventricular Remodeling

The aim of the present study was to investigate the protective effect of propofol on the experimental myocardial infarction in rats. The myocardial infarction model was established by ligating the anterior descending branch of left coronary artery in rats. Model rats were treated with propofol. Cardiac function was evaluated by echocardiography. Cardiac hemodynamic changes were detected by multiconductor biorecorder. Pathological changes in the infarcted myocardia were detected by HE staining. The expression levels of cardiac hypertrophy marker genes and fibrosis marker proteins were analyzed by real-time quantitative PCR and Western blot. The results showed that, compared with the sham surgery group, the model group exhibited larger infarct size (> 40%), impaired heart function, and significantly increased left ventricular end-diastolic pressure (LVEDP). Propofol reduced cardiac function impairment and decreased LVEDP in the model group. Propofol significantly reduced lung weight/body weight ratio, heart weight/body weight ratio, left ventricular weight/body weight ratio and left atrial weight/body weight ratio in the model group. Furthermore, after myocardial infarction, the administration of propofol significantly improved the diastolic strain rate, down-regulated the mRNA expression levels of myocardial hypertrophy markers, atrial natriuretic peptide and β-myosin heavy chain, and reversed the up-regulation of matrix metalloproteinase 2 (MMP2), MMP9 and tissue inhibitor of metalloproteinase-2 (TIMP-2) induced by myocardial infarction. These results suggest propofol can reduce adverse ventricular remodeling, cardiac dysfunction, myocardial hypertrophy and fibrosis after myocardial infarction, and has protective effect against the experimental myocardial infarction induced by coronary artery ligation in rats.
Animals ; Cardiotonic Agents/pharmacology* ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Myocardial Infarction/drug therapy* ; Myocardium ; Propofol/pharmacology* ; Rats ; Tissue Inhibitor of Metalloproteinase-2/genetics* ; Ventricular Remodeling

OBJECTIVETo explore the effect of adenovirus-mediated human stromal cell-derived factor-1alpha (hSDF-1alpha) on ventricular remodeling in rats with myocardial infarction.

METHODSA recombinant adenoviral plasmid containing hSDF-1alpha cDNA was constructed using homologous recombination in bacteria and the recombinant adenovirus particles expressing hSDF-1alpha (AdV-SDF-1) were prepared. In rat models of myocardial infarction induced by left anterior descending artery occlusion, 1x10(10) PFU AdV-SDF-1 or PFU AdV-LacZ were injected at multiple sites into the infarcted myocardium 1 h after the operation, using 200 l cell-free PBS as the control. Four weeks after the injection, the cardiac function of the rats was analyzed, and the heart tissues were taken after the measurement of hemodynamics. On serial frozen sections, histological observation and morphometric measurement were carried out using a microscopic image analysis system, and the expression of hSDF-1alpha was detected by immunocytochemistry.

RESULTSFour weeks after AdV-SDF-1 injection, the myocardium in the infracted area showed significantly higher expression rates of hSDF-1alpha. The injection resulted in a obvious reduction in the infarct size and collagen content and a marked increase in the left ventricle wall, and the rats showed improved cardiac functions.

CONCLUSIONSDF-1alpha can improve the cardiac structure and function in rats with myocardial infarction by inhibiting collagen synthesis and deposition in the infarcted area.

Adenoviridae ; genetics ; metabolism ; Animals ; Chemokine CXCL12 ; administration & dosage ; biosynthesis ; genetics ; Female ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Male ; Myocardial Infarction ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; administration & dosage ; biosynthesis ; genetics ; Transfection ; Ventricular Remodeling ; drug effects

OBJECTIVETo investigate the effects of RNA interference (RNAi) targeting angiotensin-converting enzyme (ACE) on the blood pressure and myocardial remodeling in spontaneously hypertensive rats (SHRs).

METHODSSaline (control), adenovirus (Ad5) and recombinant adenoviral vectors (Ad5-ACE-shRNA expressing ACE gene-specific shRN) were randomly administered by caudal intravasation to SHRs (n = 12 each group) at day 1 and day 16. Normotensive Wistar-Kyoto rats (WKY) served as normal controls. Systolic blood pressure (SBP) of the caudal artery was measured daily. Expressions of ACE at mRNA and protein levels in myocardium and aorta were evaluated by RT-PCR and Western blot respectively, ACE serum concentration was measured by ELISA at day 3 (n = 6 each group). The ratio of left ventricular to body weight (LVW/BW), myocardial collagen content were measured and myocardial ultrastructure observed under transmission electron microscope at the study end.

RESULTSAd5-ACE-shRNA injection significantly reduced SBP (-22 mm Hg, 1 mm Hg = 0.133 kPa) and the antihypertensive effect could last at least 14 days post each injection. SBP was not affected by saline and Ad5 injections. ACE expressions at mRNA and protein levels at myocardium and aorta as well as serum ACE were significantly decreased in Ad5-ACE-shRNA treated SHRs compared to that in saline and Ad5 groups (all P < 0.05) and was comparable to that in WKY group (P > 0.05). The LVW/BW ratio (2.24 +/- 0.19) and myocardial collagen content [(1.283 +/- 0.019) microg/mg] in Ad5-ACE-shRNA treated SHRs were also significantly lower than those in saline treated [3.21 +/- 0.13 and (1.686 +/- 0.013) microg/mg, both P < 0.05] and Ad5 treated SHRs [3.13 +/- 0.12, (1.682 +/- 0.009) microg/mg, both P < 0.05] but still higher than those of WKY group [2.06 +/- 0.11, (1.257 +/- 0.019) microg/mg, both P < 0.05]. Myocardial ultrastructure was also significantly improved in Ad5-ACE-shRNA treated SHRs compared to saline and Ad5 treated SHRs.

CONCLUSIONRNAi targeting ACE gene significantly inhibited the expressions of ACE at mRNA and protein levels and resulted in prolonged antihypertensive effects and myocardial ultrastructure improvements in this SHR model.

Animals ; Blood Pressure ; Disease Models, Animal ; Heart Rate ; Hypertension ; genetics ; physiopathology ; Male ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Ventricular Remodeling

BACKGROUNDTransplantation of adult bone marrow-derived mesenchymal stem cells (MSCs) has been proposed as a strategy for cardiac repair following myocardial damage. However cell transplantation strategies to replace lost myocardium are limited by the inability to deliver large numbers of cells that resist peritransplantation graft cell death. Accordingly, we set out to isolate and expand adult swine bone marrow-derived MSCs, and to engineer these cells to overexpress AKT1 (protein kinase B), to test the hypothesis that AKT1-engineered MSCs are more resistant to apoptosis and can enhance cardiac repair after transplantation into the ischemic swine heart.

METHODSThe CDS (regulation domain of AKT1) AKT1-cDNA fragment was amplified, and MSCs were transfected following synthesis with a pCDH1-AKT1 shuttling plasmid. Western blotting analysis and real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed. Myocardial infarction (MI) models were constructed in Meishan pigs, and cardiac function was evaluated by magnetic resonance imaging (MRI) measurements and echocardiography 4 weeks later. All pigs were assigned to four groups: control (A), DMEM (B), MSC (C), and AKT-transfected (D). MSCs were transfected with the AKT1 gene, and autologous BrdU-labeled stem cells (1 x 10(7)/5 ml) were injected into left anterior descending coronary atery (LAD) of the infarct heart in groups C and D. In group B, DMEM was injected using the same approach. In group A, there was no injection following LAD occlusion. After 4 weeks, cardiac function and regional perfusion measurements were repeated by MRI and echocardiography, and histological characteristics of the hearts were assessed. Connecxin-43 (CX-43), BrdU, and von Willebrand factor (VWF) immunoreactivity was tested using enzyme linked immunosorbent assay (ELISA). Vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGF-beta1) were analyzed at the same time.

RESULTSAKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed. AKT mRNA expression was detected at 24 hours after transfection. AKT1 expression in MSCs remained strong after 2 weeks, according to real-time RT-PCR and Western blotting. Prior to cell implantation, end-diastolic left ventricular dimension (EDLVd) increased and stroke volume (SV) decreased in the MI hearts. MRI scans revealed significantly improved cardiac function following implantation, and implanted MSCs prevented thinning and expanding in the infarct region, as well as improved contraction and increased perfusion in all groups compared to control hearts. The left ventricular chamber size was smaller in cell-transplanted hearts than in control hearts. Moreover, group D exhibited significant improvement. The expression of CX-43, BrdU, and VWF could be found in the immunohistochemical pathological sections of group C and group D. The level of VEGF reached a high level 1 week after implanting the MSCs, but the level of TGF-beta1 decreased gradually.

CONCLUSIONSThe AKT1-expressing lentiviral vector resulted in stable over-expression of AKT1 in MSCs. MSC engraftment in host myocardium improved cardiac function by attenuating contractile dysfunction and pathological thinning of the infracted left ventricular wall, which likely resulted from myocardial regeneration and angiogenesis.

Animals ; Blotting, Western ; Disease Models, Animal ; Electrophoresis ; Genetic Vectors ; genetics ; Lentivirus ; genetics ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Myocardial Infarction ; metabolism ; therapy ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine ; Ventricular Remodeling ; physiology