OBJECTIVETo investigate the microRNAs (miRNAs) expression profile of acute myocardial infarction (AMI) rats and the regulating effects of Huoxue Anxin Recipe (, HAR) on angiogenesis-related miRNAs and genes.

METHODSForty-five Wistar rats were randomly assigned to 3 groups according to a random number table: sham, AMI, and AMI+HAR groups (15 in each group). AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI+HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miRNAs expression profile was detected by quantitative realtime polymerase chain reaction (qRT-PCR). The mRNA and protein expressions of vascular endothelial growth factor (VEGF), and a target gene of miR-210 was further detected by qRT-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining.

RESULTSCompared with the sham group, ejection fraction (EF) and fractional shortening (FS) values were decreased significantly in the AMI group (P<0.01), while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI+HAR group, EF and FS values were increased significantly (P<0.05 vs. AMI group), and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miRNAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miRNAs in the AMI+HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI+HAR group. The relative mRNA and protein expressions of VEGF were decreased significantly in the AMI group (P<0.05 vs. sham group), and increased significantly in the AMI+HAR group (P<0.01 vs. AMI group). CD31 expression area and optical intensity were decreased significantly in the AMI group (P<0.05 vs. sham group), and increased significantly in the AMI+HAR group (P<0.01 vs. AMI group).

CONCLUSIONSHAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Heart Function Tests ; Male ; MicroRNAs ; genetics ; metabolism ; Microvessels ; pathology ; Myocardial Infarction ; drug therapy ; genetics ; physiopathology ; Myocardium ; pathology ; Neovascularization, Physiologic ; drug effects ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats, Wistar ; Up-Regulation ; drug effects ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

PURPOSE: Adipose-derived stem cells (ADSCs) are known to be potentially effective in regeneration of damaged tissue. We aimed to assess the effectiveness of intracoronary administration of ADSCs in reducing the infarction area and improving function after acute transmural myocardial infarction (MI) in a porcine model. MATERIALS AND METHODS: ADSCs were obtained from each pig's abdominal subcutaneous fat tissue by simple liposuction. After 3 passages of 14-days culture, 2 million ADSCs were injected into the coronary artery 30 min after acute transmural MI. At baseline and 4 weeks after the ADSC injection, 99mTc methoxyisobutylisonitrile-single photon emission computed tomography (MIBISPECT) was performed to evaluate the left ventricular volume, left ventricular ejection fraction (LVEF; %), and perfusion defects as well as the myocardial salvage (%) and salvage index. At 4 weeks, each pig was sacrificed, and the heart was extracted and dissected. Gross and microscopic analyses with specific immunohistochemistry staining were then performed. RESULTS: Analysis showed improvement in the perfusion defect, but not in the LVEF in the ADSC group (n=14), compared with the control group (n=14) (perfusion defect, -13.0+/-10.0 vs. -2.6+/-12.0, p=0.019; LVEF, -8.0+/-15.4 vs. -15.9+/-14.8, p=0.181). There was a tendency of reducing left ventricular volume in ADSC group. The ADSCs identified by stromal cell-derived factor-1 (SDF-1) staining were well co-localized by von Willebrand factor and Troponin T staining. CONCLUSION: Intracoronary injection of cultured ADSCs improved myocardial perfusion in this porcine acute transmural MI model.
Adipose Tissue/cytology ; Animals ; Bone Marrow Cells/cytology/*metabolism ; Chemokine CXCL12 ; Coronary Vessels ; Female ; Heart/physiopathology ; Heart Ventricles ; *Mesenchymal Stromal Cells ; Myocardial Infarction/physiopathology/radionuclide imaging/*therapy ; *Stem Cell Transplantation ; Swine ; Technetium Tc 99m Sestamibi/*pharmacology ; Tomography, Emission-Computed, Single-Photon/*methods ; Troponin T ; *Ventricular Function, Left

OBJECTIVETo study the effect of medicines for activating blood and reinforcing Qi on the number of new micro-vessels and the protein expressions of VEGF and bFGF in the infarcted myocardium edge area of acute myocardial infarction (AMI) model in rats.

METHODThe AMI model of rats was established. After the successful model establishment, rats were randomly divided into the sham-operated group, the model group, the Danshen-Huangqi (1 : 2) group, the Danshen-Huangqi (1 : 1) group, the Chuanxiong-Huangqi (1 : 2) group, the Danshen group, the Chuanxiong group, the Chishao group and the Shexiang Baoxin pill group, with five rats in each group. Rats in each medicated group were orally administered with drugs as per 13.5 g x kg(-1) x d(-1) once everyday for three weeks. The immunohistochemical SP method was adopted to detect the expression of vWF in myocardial tissues, and count the number of micro-vessels (MVC). The protein expression of VEGF and bFGF in myocardial tissues were determined by Western blot.

RESULTThe new micro-vessels stained by vWF factor could be found in the infarcted myocardium edge area of the sham-operated group, the model group and all of medicated groups. The sham-operated group show unobvious new micro-vessels in myocardial tissues. A small amount of new micro-vessels could be seen in the infarcted myocardium edge area of the model group. Whereas a larger number of micro-vessels could be seen in the infarcted myocardium edge area of all of medicated groups. The differences between the sham-operated group and the model group had statistical significance (P < 0.05). The differences between each medicated group and the model group had statistical significance as well (P < 0.05 or P < 0.01). The lowest protein expression of VEGF and bFGF was found in myocardium of the sham-operated group, with the statistical significance compared with the model group (P < 0.05). Compared with the model group, each medicated group showed significant increase in the protein expression of VEGF and bFGF, with the statistical significance between them (P < 0.05 or P < 0.01).

CONCLUSIONThe Danshen group, the Chuanxiong group, the Chishao group, the Danshen-Huangqi (1 : 2) group, the Danshen-Huangqi (1 : 1) group and the Chuanxiong-Huangqi (1 : 2) group show the effect in promoting angiogenesis. Their mechanism for promoting angiogenesis may be related to the improvement of the protein expressions of VEGF and bFGF, so as to increase the contents of VEGF and bFGF and promote the angiogenesis of new vessels.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Humans ; Male ; Microcirculation ; drug effects ; Microvessels ; drug effects ; physiopathology ; Myocardial Infarction ; drug therapy ; physiopathology ; Neovascularization, Pathologic ; drug therapy ; genetics ; metabolism ; Qi ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVE: To determine the improvements of post-infarction heart function after transplantation of autologous skeletal myoblasts transfected with VEGF165 in rabbits. METHODS: Myocardium infarction was induced in rabbits by left anterior descending coronary artery ligation. At 2 weeks, 1.75×10(7) autologous skeletal myoblasts transfected with pcDNA3.1-VEGF165 were infused into the region of MI via direct intramuscular injection; pcDNA3.1 served as a control. RESULTS: The DAPI-labeled and Desmin-positive immunostained skeletal myofibers were found throughout the infracted areas and border zones, and the density of blood capillary in the MI region transplanted by myoblasts with VEGF165 was increased (measured 4 weeks later and compared with controls). Heart function was examined by the Buxco system and demonstrated that maximum dp/dt [(1607.23±102.67) mmHg/s vs (1217.77±89.91) mmHg/s] and minimum dp/dt [(-1535.09 ± 81.34) mmHg/s vs (1174.58 ± 91.5) mmHg/s] were improved in the heart transplanted with the transfected myoblasts(P<0.05). CONCLUSION Autologous skeletal myoblasts transfected with VEGF165 could ameliorate the blood supply in the MI region, and aid recovery of heart function more quickly in post-infarction hearts. This suggests an effective treatment for myocardium infarction.
Animals ; Female ; Myoblasts, Skeletal ; metabolism ; transplantation ; Myocardial Infarction ; physiopathology ; therapy ; Rabbits ; Recovery of Function ; Transfection ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Ventricular Function, Left ; physiology

Cardiac lymphatic system in the remodeling after acute myocardial infarction (AMI) has been overlooked. We wanted to investigate the role of bone marrow-derived endothelial progenitor cells (EPCs) and their contribution to lymphatic distribution in myocardial remodeling after AMI. Mouse (C57bl/6J) MI models were created by ligation of the left anterior descending coronary artery and were treated with phosphate buffered saline (PBS) or EPCs. Real-time RT-PCR with 2- to 4-week myocardial tissue samples revealed that lymphangiogenetic factors such as vascular endothelial growth factor (VEGF)-C (8.5 fold, P < 0.05), VEGF-D (6.1 fold, P < 0.05), Lyve-1 (15 fold, P < 0.05), and Prox-1 (11 fold, P < 0.05) were expressed at significantly higher levels in the PBS group than the EPC group. The PBS group also showed a significantly higher density of lymphatic vessels in the peri-infarction area. Echocardiography showed that from 2 weeks after the treatment, left ventricle (LV) dimensions at both systole and diastole were significantly smaller in the EPC group than in the PBS group (P < 0.01) and LV fractional shortening was higher in the EPC group accordingly (P < 0.01). Lymphangiogenic markers increased in a mouse MI model. EPC transplantation decreased lymphangiogenesis and adverse ventricular remodeling after AMI. These novel findings suggest that new lymphatic vessels may be formed in severely damaged myocardium, and may be involved in adverse myocardial remodeling after AMI.
Animals ; Cell Transplantation ; Endothelial Cells/*cytology ; Homeodomain Proteins/genetics/metabolism ; Immunohistochemistry ; Lymphangiogenesis/genetics/*physiology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Myocardial Infarction/metabolism/physiopathology/*therapy ; Real-Time Polymerase Chain Reaction ; *Stem Cell Transplantation ; Tumor Suppressor Proteins/genetics/metabolism ; Vascular Endothelial Growth Factor A/genetics/metabolism ; Vascular Endothelial Growth Factor D/genetics/metabolism

OBJECTIVETo study the changes of cardiac function following treatment with granulocyte colony stimulating factor (G-CSF) in patients with heart failure after myocardial infarction.

METHODSThirty-eight patients with heart failure after myocardial infarction were randomized into G-CSF treatment group and control group. All the patients received conventional treatment (medication and interventional therapy), and the patients in treatment group were given additional G-CSF (600 µg/day) for 7 consecutive days. The plasma level of brain-type natriuretic peptide (BNP) and the number of endothelial progenitor cells (EPCs) in the peripheral blood were detected before and at 7 days and 4 months after the treatment. The cardiac functions (LVSD, EDV, and LVEF) were evaluated by ultrasonic imaging before and at 2 weeks and 4 months after the treatment.

RESULTSThe number of EPCs was significantly higher in the treatment group than in the control group after the treatment especially at 7 days (P<0.01). In both groups, BNP level was lowered significantly after the treatment to recover the normal level (P<0.01). The cardiac functions were improved in all the patients at 7 days and 4 months after the treatment, and the improvement was more obvious in the treatment group (P<0.05), especially in terms of LVEF at 4 months after the treatment (P<0.01).

CONCLUSIONEPC mobilization by G-CSF can effectively improve the cardiac functions and lessen ventricular remodeling in patients with heart failure after myocardial infarction.

Aged ; Endothelial Cells ; cytology ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Heart Failure ; etiology ; physiopathology ; therapy ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Myeloid Progenitor Cells ; cytology ; Myocardial Infarction ; complications ; physiopathology ; therapy ; Natriuretic Peptide, Brain ; metabolism ; Treatment Outcome ; Ventricular Remodeling

PURPOSE: Ethyl pyruvate has anti-inflammatory properties and protects organs from ischemia/reperfusion (I/R)-induced tissue injury. The aim of this study was to determine whether ethyl pyruvate decreases the inflammatory response after regional I/R injury and whether ethyl pyruvate protects against delayed regional I/R injury in an in vivo rat heart model after a 24 hours reperfusion. MATERIALS AND METHODS: Rats were randomized to receive lactated Ringer's solution or ethyl pyruvate dissolved in Ringer's solution, which was given by intraperitoneal injection 1 hour prior to ischemia. Rats were subjected to 30 min of ischemia followed by reperfusion of the left coronary artery territory. After a 2 hours reperfusion, nuclear factor kappaB, myocardial myeloperoxidase activity, and inflammatory cytokine levels were determined. After the 24 hours reperfusion, the hemodynamic function and myocardial infarct size were evaluated. RESULTS: At 2 hours after I/R injury, ethyl pyruvate attenuated I/R-induced nuclear factor kappaB translocation and reduced myeloperoxidase activity in myocardium. The plasma circulating levels of inflammatory cytokines decreased significantly in the ethyl pyruvate-treated group. At 24 hours after I/R injury, ethyl pyruvate significantly improved cardiac function and reduced infarct size after regional I/R injury. CONCLUSION: Ethyl pyruvate has the ability to inhibit neutrophil activation, inflammatory cytokine release, and nuclear factor kappaB translocation. Ethyl pyruvate is associated with a delayed myocardial protective effect after regional I/R injury in an in vivo rat heart model.
Animals ; Anti-Inflammatory Agents/*pharmacology ; Cell Nucleus/metabolism ; Cytoplasm/metabolism ; Heart/physiopathology ; Inflammation ; Male ; Myocardial Infarction/prevention & control ; Myocardium/*metabolism ; NF-kappa B/metabolism ; Peroxidase/metabolism ; Pyruvates/*pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury/*drug therapy/*metabolism

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 evaluate the protective effects of 8% emulsified isoflurane after myocardial ischemia-reperfusion injury and its mechanism in rabbits.

METHODSTwenty-four male adult New Zealand white rabbits were anesthetized with intravenous injection of 30 mg/kg pentobarbital followed by 5 mg x kg(-1) x h(-1) infusion. All rabbits were subjected to 30 minutes of left anterior descending coronary artery (LAD) occlusion and 3 hours of subsequent reperfusion. Before LAD occlusion, the rabbits were randomly allocated into three groups for preconditioning treatment (eight for each group). The control group (C group) received intravenously 0.9% NaCl for 30 minutes. The emulsified isoflurane group (EI group) received 8% emulsified isoflurane intravenously till 0.64% end-tidal concentration for 30 minutes that was followed by a 15-minute washout period. The Intralipid group (IN group) received 30% Intralipid for 30 minutes. The infarcted area, plasma malondialdehyde (MDA) content, superoxide dismutase activity (SOD) and nitrite concentration after 3-hour myocardial perfusion were recorded simultaneously.

RESULTSFor the myocardial ischemia-reperfusion injury animals, the infarcted size in the EI group was significantly reduced (91.9% +/- 8%) as compared with control group (39% +/- 6%, t=5.19, P<0.01). The plasma SOD activity and nitrite concentration in EI group were significantly higher than those in control group (t=2.82, t=8.46, P<0.05), but MDA content was lower in EI group than that in control group (t=2.56, P<0.05).

CONCLUSIONSThe results indicate that emulsified isoflurane has a cardioprotection effect against ischemia-reperfusion injury. This beneficial effect of emulsified isoflurane is probably through NO release and consequently by increase in antioxidation of myocardium.

Animals ; Emulsions ; Isoflurane ; administration & dosage ; pharmacology ; Lipid Peroxidation ; drug effects ; Male ; Myocardial Infarction ; drug therapy ; pathology ; Myocardial Reperfusion Injury ; physiopathology ; prevention & control ; Nitric Oxide ; blood ; Rabbits ; Superoxide Dismutase ; metabolism

OBJECTIVETo observe the effects of Panax notoginsenoside (PNS) on tumor necrosis factor-alpha (TNF-alpha) and matrix metalloproteinases-2 (MMP-2) expressions in rats with post-myocardial infarction ventricular remodeling and explore the mechanism.

METHODSRat models of acute infarction ventricular (AMI) were established by ligation of the left anterior descending coronary artery. Twenty-four hours after the operation, the rats were randomized into control and experimental groups for intragastric administration of normal saline (control), fosinopril and PNS at the low, medium and high doses for 4 consecutive weeks. The effects of PNS on the cardiac function index including the left ventricular end-diastolic dimension (LVIDd), left ventricular end-systolic diameter (LVIDs), ejection fraction (EF), percentage of left ventricular systole (FS), mitral early diastolic flow velocity mouth (MV), and heart rate (HR) were observed, and the changes in TNF-alpha and MMP-2 expression were detected after post-myocardial infarction ventricular remodeling.

RESULTSCompared with the control group, PNS at the medium and high doses produced significant improvements in the EF, FS and MV of the rats (P<0.01 or 0.05). TNF-alpha and MMP-2 expressions were significantly decreased by PNS treatment at low, medium and high doses (P<0.01).

CONCLUSIONPNS can inhibit or reduce the expression of TNF-alpha and MMP-2, thereby enhancing left ventricular systolic and diastolic functions, decreasing peripheral resistance, and improving the cardiac function of rats with post-myocardial infarction left ventricular remodeling.

Animals ; Female ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Myocardial Infarction ; drug therapy ; physiopathology ; Panax notoginseng ; chemistry ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; therapeutic use ; Tumor Necrosis Factor-alpha ; metabolism ; Ventricular Remodeling ; drug effects