Long-term effects of TCV116 on left ventricular remodeling and heart function after myocardial infarction in rats

AIM: To investigate the effects of long-term TCV116 on left ventricular remodeling and heart function after myocardial infarction. METHODS: Myocardial infarction (MI) was caused by ligation of the left anterior descending coronary artery in rats. One week after the surgical performance, the surviving rats were randomly assigned to the following treatment protocols: (1) MI rats with no therapy; (2) MI rats treated with TCV116 2 mg/kg per day; (3) Sham-operated control; (4) Sham-operated rats, treated with TCV116 2 mg/kg per day. At 22 weeks, cardiac hemodynamic parameters such as MAP, LVSP, dp/dt_max and LVEDP, and histomorphometric parameters such as LVW/BW and LVCA/BW were measured, mRNA of cardiac genes such as βMHC, BNP, TGF-β₁, collagen I and III were quantified, and survival rates were calculated. RESULTS: Compared with sham-operated rats, MI rats without therapy showed significant increases in histomorphometric parameters as well as in mRAN expressions of cardiac genes (P<0.01); While their hemodynamic parameters were significantly impaired (P<0.01), and survival duration shortened (P<0.05). Compared with MI rats without therapy, MI rats treated with TCV116 showed significant attenuation of mRAN expression of cardiac genes (P<0.01); While their hemodynamic parameters were significantly improved (P<0.05 or P＜0.01), and survival duration extended (P<0.05). CONCLUSION: Treatment with long-term angiotensin II type 1 receptor antagonist may improve left ventricular remodeling and cardiac function after MI in rats.     

Role of PPARα/PGC-1α in doxorubicin induced mouse dilated cardio-myopathy

AIM: To investigate the changes of peroxisome proliferator-activated receptors (PPAR)α/peroxisome proliferator activated receptor coactivator 1 alpha (PGC-1α) in doxorubicin (DOX) induced dilated cardiomyopathy (DCM) and its effect on the energy metabolism and myocardial function in mice. METHODS: Forty mice were randomly divided into 4 groups: control group, DOX group, PPARα inhibitor group and PPARα agonist group. The DCM model was established by injection of DOX. The protein levels of PPARα/PGC-1α were detected. The PPARα inhibitor and PPARα agonist were used 2 weeks beforeinjection of DOX. The contents of adenine acid and phosphocreatine (Pcr) in the mitochondria were measured by high-performance liquid chromatography (HPLC). The ANT activity was analyzed by the atractyloside-inhibitor stop technique. The changes of the echocardiography and hemodynamics were also observed. RESULTS: DOX induced DCM model was successfully established. The protein levels of PPARα and PGC-1α in control group were significantly higher than those in DOX group (P<0.05). Both of the high-energy phosphate contents and the transport activity of ANT were decreased in DOX group (P<0.05), and the hemodynamic parameters were disordered (P<0.01). Compared with DOX group, PPARα inhibitor pre-treatment significantly reduced the PPARα/PGC-1α expression. Meanwhile, high-energy phosphate contents in the mitochondria and the ANT transport activity of the mitochondria decreased, as well as the left ventricular function (P<0.05). On the other hand, PPARα agonist significantly increased the expression of PPARα and PGC-1α, and improved the transport activity of ANT. In addition, the hemodynamic parameters were ameliorated, but the high-energy phosphate contents of the mitochondria did not significantly change. CONCLUSION: PPARα/PGC-1α plays an important role in the regulation of ANT transport activity in dilated cardiomyopathy induced by DOX, and the activation of PPARα/PGC-1α has protective effects on the DCM induced by DOX.     

Effects of noninvasive delayed limb ischemic preconditioning on prognosis of myocardial infarction

AIM: To study the effects of noninvasive delayed limb ischemia preconditioning (NDLIP) on animal cardiac function, myocardial morphology and myocardial apoptosis after myocardial infarction (MI). METHODS: Healthy SD male rats[n=45, weighing (250±10) g] were randomly divided into 3 groups:MI group:the animal model of MI was established by surgical ligation of left anterior descending artery (LAD) after 2 weeks; NDLIP group:after the success of the MI animal model, NDLIP was carried out every other day until the 4th, 6th and 8th weeks; sham group:as the negative control group, the animals were taken heart LAD threading but no ligation. All rats were fed conventionally. At the end of the 4th, 6th and 8th weeks, all rats were made ventricular intubation, and then the hemodynamic parameters were recorded. The blood samples were withdrawn from the abdominal aorta and the serum was separated via centrifugation. The serum contents of Bcl-2 and Bax were measured by ELISA. Left ventricular anterior wall was homogenized. The mitochondrial respiratory chain complexes Ⅰ, Ⅱ, Ⅲ and Ⅳ in the myocardial tissues were detected by ELISA. RESULTS: At the end of the 4th, 6th and 8th weeks, compared with MI group, left ventricular systolic pressure in NDLIP group was significantly increased, while left ventricular end-diastolic pressure in NDLIP group was significantly decreased (both P<0.05). Mitochondrial respiratory chain complexesⅠ, Ⅱ, Ⅲ and Ⅳ in NDLIP group were significantly increased (P<0.05). The serum level of Bcl-2 in NDLIP group was significantly increased and Bax level was reduced remarkably (both P<0.01). CONCLUSION: NDLIP improves the hemodynamic indexes, promotes the mitochondrial respiratory function and inhibits cell apoptosis, thus improving the prognosis of MI.     

Effect of angelica on myocardial fibrosis post myocardial infarction in rats

AIM: To investigate 1) the role of transforming growth factor-β₁ (TGF-β₁) and macrophage infiltration during the development of myocardial fibrosis (MF) in rats after myocardial infarction (MI);and 2) mechanisms of MF post-MI and the inhibitory effect of angelica.METHODS: Sprague-Dawley (SD) rats were subjected to MI by ligating the left anterior descending coronary artery.The animals were randomly divided into three groups: sham, MI and MI+angelica.After 24 hours of ligation, rats received angelica (20 mL·kg^-1·d^-1, ip) or saline.Left ventricular hemodynamics were measured and rats were killed at week 1, week 2 and week 4, respectively.Collagen content, macrophage infiltration and TGF-β₁ expression were examined in the non-infarcted area.RESULTS: ① In MI group, the numbers of macrophage and TGF-β₁ expression were significantly upregulated compared to sham at week 1 post-MI and remained elevated at week 4 (P<0.01).Angelica significantly decreased macrophage infiltration and TGF-β₁ expression (P<0.01 vs MI).② Collagen content was increased significantly in MI group compared to sham at week 2 and week 4 (P<0.01), and decreased in MI+angelica group (P<0.05 vs MI).③ Cardiac function was markedly decreased post-MI in MI group (P<0.01), and improved at week 4 in MI+angelica group (P<0.05).CONCLUSION: In MF post-MI, angelica may have an antifibrotic effect by decreasing macrophage infiltration and TGF-β₁ expression, by which reactive myocardial fibrosis is reduced, and cardiac function is improved.     

Effect of pioglitazone on pre-adipocytes differentiation and GILZ expression

AIM: To investigate the roles of pioglitazone on differentiation and expression of GILZ in 3T3-L1 pre-adipocytes. METHODS: The morphological changes during 3T3-L1 cell differentiation were observed. The cells were treated with pioglitazone at concentrations of 1×10^-4～1×10^-2 mmol/L for 48 h, then the relative content of triglyceride were analyzed by oil red O staining at 2nd, 4th and 6th day during adipogenesis. The mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2) and lipoprotein lipase (LPL) was measured by real-time PCR. GILZ protein expression was determined by Western blot after the cells were treated with pioglitazone at concentrations of 1×10^-4 ～1×10^-2 mmol/L for 48 h.RESULTS: Oil-red O staining showed that the relative contents of triglyceride in adipocytes were increased with the increase in the pioglitazone concentration. Compared with the control, the relative contents of triglyceride in group 1×10^-3 mmol/L and group 1×10^-2 mmol/L were significantly increased (P < 0.05). The mRNA expression of PPARγ2 and LPL was also increased with the increase in the pioglitazone concentration. When pioglitazone concentration was more than 1×10^-3 mmol/L, compared with the control, the mRNA expression of PPARγ2 and LPL significantly increased (P < 0.01). The protein expression of GILZ was decreased with the increase in the pioglitazone concentration.CONCLUSION: Pioglitazone down-regulates GILZ expression, and up-regulate PPARγ2 expression and the downstream functional factor such as LPL.     

Salvia extract promotes angiogenesis of myocardium in rats with myocardial infarction

AIM: To determine the effect of salvia extract on angiogenesis of the myocardium in the rats with myocardial infarction (MI) and to analyze its possible mechanism. METHODS: Left coronary artery of Sprague-Dawley rats was ligated to establish a MI model. The rats were randomly divided into MI model group, 3 different dose groups of salvia (10, 20 and 40 mg·kg^-1·d^-1), and sham operation group. Each group consisted of 8 rats. The rats in all treatment groups were orally administered with the salvia extract, and the rats in MI group and sham operation group were fed with the same volume of saline. The rats were sacrificed 4 weeks later. The hemodynamic changes of the rats were determined, and the segmental heart samples were used for morphological observation by hematoxylin and eosin staining, Masson staining, or electron microscopic analysis. The expression of vascular endothelial growth factor (VEGF) and cluster of differentiation 34 (CD34) was analyzed according to immunohistochemistry. RESULTS: Compared with sham operation group, the morphological changes of the myocardium in MI group were disordered, part of myocardial cell outline disappeared, and obvious fibrosis in the necrosis myocardial tissue and fuzzy or disappearing microvascular ultrastructure were also observed. Compared with MI group, the number of new microvessels in all the treatment groups increased obviously, and the morphological changes of the endothelial cells were relatively complete according to electron microscopy. Compared with sham operation group, the protein expression of VEGF and CD34 in the cytoplasm of the myocardial tissues in MI group increased only a little. Compared with MI group, the protein expression of VEGF and CD34 in the cytoplasm of the myocardial tissues in all treatment groups increased significantly (P<0.01). CONCLUSION: Salvia extract obviously promotes angiogenesis of the myocardial tissues in the rats after myocardial infarction.     

Effects of celecoxib on cardiac myocyte apoptosis after myocardial infarction

AIM: To investigate the effects of celecoxib, a selective cyclooxygenase-2 inhibitor, on antioxidative capability and apoptosis of cardiac myocytes after myocardial infarction. METHODS: 24 New Zealand rabbits were divided into three groups randomly (8 in each group): sham-operated group (sham group), myocardial infarction group (MI group), celecoxib group (Cele group, 10 mg kg^-1·d^-1, qd, with the drugs gastric gavage for six weeks). The NO concentration, total antioxidative capability (T-AOC), the activity of constitutive nitric oxide synthase (cNOS) and inducible NOS (iNOS) in cardiac tissue homogenate, adjacent to the infracted area, were detected. The pathological changes were observed by light microscope and electron microscopy. The expressions of Bcl-2 and Bax protein in myocytes were observed using immunohistochemistry, and the degree of apoptosis were examined by TUNEL. RESULTS: Cardiac tissue in MI group presented interstitial edema, fibroplastic proliferation, inflammatory cellular infiltration, and vacuolar degeneration in cardiac myocytes. The results of electron microscopy showed that myocytes presented more changes caused by ischemic injury: widened interspace of myofibril, disordered myofibrillae, focal lysis of myofilament, ectasia of sarcoplasmic reticulum. In Cele group, the pathological changes were light, the NO^-₂/ NO^-₃ concentration, the activity of iNOS were lower (P<0.05), while the activity of cNOS and T-AOC were higher (P<0.05) than those in MI group. The expression rate of Bcl-2 protein in Cele group was higher than that in MI group, while the Bax was lower (P<0.01). The number of apoptotic myocytes was lower than that in MI group (P<0.01).CONCLUSION: Celecoxib decreases the number of apoptotic cardiomyocytes and increases the antioxidative capability after myocardial infarction.     

Effects of soluble transforming growth factor-β type Ⅱ receptor on cardiac functions after myocardial infarction in rats

AIM: To observe the effects of soluble transforming growth factor-β type Ⅱ receptor (sTGFβRⅡ) on cardiac functions after myocardial infarction (MI) in rats. METHODS: MI was induced in Sprague-Dawley (SD) rats by ligating the left anterior descending coronary artery. The rats surviving to the third day after MI were included in the study and randomly divided into MI group, pAd-sTGFβRⅡ group (transfected with recombinant adenovirus vector expressing the extracellular domain gene of TGF-βRⅡ), vector group and sham group. Four weeks later, the heart rate (HR), left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic dimension (LVESD) and ejection fraction (EF) were evaluated by echocardiograms. The expression of sTGFβRⅡ in myocardial tissues was observed under fluorescence microscope by frozen sectioning, and the expression of typeⅠ and Ⅲ collagens was observed by Sirius red-saturated picric acid staining. The expression of matrix metalloproteinase 9 (MMP-9) at mRNA and protein levels was determined by RT-PCR and immunohistochemical method. The activity of MMP-9 was assayed by gelatin zymography. RESULTS: Compared with sham group, HR, LVEDD, LVESD, typeⅠ and Ⅲ collagen, mRNA and protein of MMP-9, and the activity of MMP-9 increased significantly (P<0.01), and EF decreased (P<0.01) in MI group and vector group. Compared with MI group, EF was increased (P<0.01), but HR, LVEDD, LVESD, typeⅠ and Ⅲ collagen, mRNA and protein expression of MMP-9 and the activity of MMP-9 decreased significantly (P<0.01) in pAd-sTGFβRⅡ group, and all the parameters above were still higher than those in sham group. CONCLUSION: sTGFβRⅡ intervention improves the cardiac functions after MI by inhibiting TGF-β-mediated MMP-9 expression.     

Changes of cytokine and collagen in the myocardial remodeling after acute myocardial infarction in rats

AIM:To clarify the relationship between the cytokine and collagen in myocardial remodeling after acute myocardial infarction (MI) in rats. METHODS:In MI group, Wistar rats were undergone acute myocardial infarction by ligation of the anterior descending coronary artery. Sham operation was made in rats as control. The mRNA expression of collagen and cytokines such as TNF-α and TGF-β1 in infract and non-infarct region of left myocardium were detected by RT-PCR at different time point (3 d, 1 and 4 weeks). RESULTS:Collagen type Ⅰ and Ⅲ elevated as well as the TNF-α and TGF-β1 in the MI group at 3th day. Expression of collagen type Ⅰ and Ⅲ were higher in the infarct region than that in the non-infarct region even at 4 weeks. TNF-α and TGF-β1 peaked at 1 week and declined gradually to the baseline, which was still higher than those in control group (P<0.01). Correlation analysis revealed that expressions of TNF-α and TGF-β1 were positively correlated with the collagen type Ⅰand Ⅲ (P<0.01). CONCLUSION:Cytokines participate in the myocardial remodeling after MI. Interfering with expression of cytokines may be the potentially preventative method in the myocardial remodeling.     

Effect of hydrogen sulfide on myocardial fibrosis and PPARγ and NF-κB expression in rat model of diabetes

AIM: To explore the effects of hydrogen sulfide (H₂S) on the myocardial fibrosis in a rat model of diabetes and its mechanism.METHODS: Single intraperitoneal injection of streptozotocin (STZ) was utilized to establish a rat model of diabetes. Sodium hydrosulfide was used as an exogenous donor of hydrogen sulfide. Male SD rats were randomly divided into control group, STZ group, STZ+H₂S group and H₂S group. Eight weeks later, HE and VG staining methods were used to observe the collagen distribution and collagen volume fraction was measured by image analysis. The expression levels of type I collagen, PPARγ and NF-κB in the cardiac tissues were determined by Western blotting.RESULTS: Compared with control group, collagen distribution and the expression levels of type I collagen and NF-κB in the cardiac tissues were markedly increased (P<0.05), while PPARγ was significantly decreased in STZ group (P<0.05), but these indexes were reversed significantly in STZ+H₂S group (P<0.05). The expression levels of type I collagen, PPARγ and NF-κB had no significant difference between H₂S group and control group.CONCLUSION: Hydrogen sulfide attenuates cardiac fibrosis in diabetic rats, and its mechanism may be related to PPARγ-NF-κB signaling pathway.     

Effects of pioglitazone on cognitive impairments induced by isoflurane anesthesia in aged mice

AIM：To investigate the effects of pioglitazone, a peroxisome proliferator-activated receptor γ (PPARγ) agonist, on the cognitive impairments and inflammatory cytokine production induced by isoflurane in aged mice. METHODS：Male C57BL/6J mice (11-month-old, n=136) were assigned randomly into 5 groups: control group (Con), isoflurane group (Iso), 10 mg/kg pioglitazone + isoflurane group (Pi10+Iso), 20 mg/kg pioglitazone + isoflurane group (Pi20+Iso) and 20 mg/kg pioglitazone alone group (Pi20). The mice in all isoflurane-treated groups were exposed to oxygen mixed with 1.4% isoflurane for 2 h. The mice in Con group and in Pi20 group were exposed to oxygen only for 2 h. Pioglitazone was suspended in 1% carboxymethyl cellulose (CMC). Pioglitazone (10 mg/kg or 20 mg/kg) was gavaged 2 h prior to the exposure of isoflurane or oxygen alone. The same volume of 1% CMC was gavaged in Con group and in Iso group. Fear conditioning tests were performed to determine the learning and memory abilities 48 h after isoflurane exposure. Fresh cerebral cortice and hippocampi were dissected to measure the protein expression of PPARγ by Western blotting, and the contents of IL-1β and TNF-α were analyzed by ELISA 6 h after isoflurane exposure. RESULTS：Compared with Con group, the response of freezing behavior decreased (P<0.05) and IL-1β content in the hippocampus increased (P<0.05) in Iso group. Compared with Iso group, the response of freezing behavior and PPARγ protein expression level had no significant change (P>0.05) in Pi10+Iso group, but the response of freezing behavior and PPARγ protein expression level increased significantly (P<0.05) and IL-1β content in the hippocampus decreased (P<0.05) in Pi20+Iso group. IL-1β content in the cerebral cortex and TNF-α levels both in the cerebral cortex and the hippocampus showed no significant difference among all groups (P>0.05). CONCLUSION：Pioglitazone attenuates cognitive impairments and the elevates the level of IL-1β in the hippocampus induced by isoflurane in aged mice.     

Effect of erythropoietin on expression of myocardial NADPH oxidase in pressure overload rats

AIM:To explore the effect of erythropoietin （EPO） on the expression of myocardial NADPH oxidase （Nox） in the pressure overload rats. METHODS:Male SD rats (n=36) were used to establish a pressure overload myocardial hypertrophy model by abdominal aorta ligation. The animals were divided into model group, control group (sham, without narrowing abdominal aorta, the rest of the operation was the same as the model) and recombinant human erythropoietin (rhEPO) treatment group (intraperitoneal injection of rhEPO postoperatively, 4 000 U/kg, twice a week). After 8 weeks, the cardiac ultrasound imaging and hemodynamic evaluation were conducted to determine the cardiac functions. Masson staining was used to observe the degree of myocardial fibrosis. The expression of Nox2 and Nox4 at mRNA and protein levels was detected by real-time quantitative PCR and Western blotting. The protein levels of myocardial inflammatory factors CD45, F4/80 and TGF-β were determined by Western blotting. RESULTS:Compared with model group, the left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), left ventricular end-systolic pressure (LVESP) and left ventricular pressure maximum rising and falling rates (±dp/dtmax) increased significantly in EPO treatment group (P<0.01). At the same time, left ventricular end-systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDD) and left ventricular end-diastolic pressure (LVEDP) were decreased in EPO treatment group (P<0.01). EPO reduced the degree of myocardial fibrosis caused by pressure overload (P<0.01) and decreased the expression of Nox2 and Nox4 at mRNA and protein levels in the myocardium (P<0.05 or P<0.01), and reduced the protein expression of myocardial inflammatory factors CD45, F4/80 and TGF-β. CONCLUSION: EPO inhibits rat myocar-dial fibrosis induced by pressure overload, improves heart functions by decreasing NADPH oxidase activity and inhibiting myocardial oxidative stress levels and myocardial inflammatory reaction.     

Changes of calcium handling protein after acute myocardial infarction in rats and effect of carvedilol

AIM: To observe the changes of sarcoplasmic reticulum Ca2+-ATPase (SERCA)， phospholamban (PLB) during heart failure after acute myocardial infarction (AMI) in rats and the effect of carvedilol. METHODS: Rats were randomly assigned to normal control group, sham-operation group, AMI group and carvedilol (CAR) group. 6 weeks later, in vivo hemodynamic, morphometry and SERCA, PLB mRNA and protein expression of myocytes were measured in all animals. RESULTS: In comparison with sham-operation group, LV end diastolic pressure (LVEDP) and weight of ventricles were increased, while maximal rate of rise and fall (±dp/dt) of LV pressure were decreased in AMI group. After treatment with carvedilol, these parameters were all improved. The mRNA and protein expression of SERCA were downregulated (P<0.01). PLB mRNA and protein expression were upregulated (P<0.01) in AMI group relative to sham-operation group. Carvedilol restored the low expression of SERCA mRNA and protein (P<0.05), but was no effect on PLB mRNA and protein expression (P>0.05). CONCLUSIONS: The changes of SERCA and PLB may be the important mechanism of contractile dysfunction in heart failure after AMI. Carvedilol is effective in preventing LV dysfunction after AMI. The molecular mechanism may be related with normalization of SERCA expression.     

Role of benazepril on extracellular signal-regulated kinase activity and expression of B-type natriuretic peptide in spontaneously hypertensive rats

AIM：To investigate the role of benazepril on extracellular signal-regulated kinase (ERK) activity and expression of B-type natriuretic peptide in spontaneously hypertension rat (SHR).METHODS：Wistar Kyoto rats were used as control group.Twenty one 14-week-age SHR were randomized into 3 groups,7 rats each：benazepril group (10 mg·kg-1·d-1);hydralazine group (10 mg·kg-1·d-1) and sham group.In each group drugs or equal volume of vehicle (0.5% carboxymethyl cellulose) were administered respectively for 10 weeks by gavage.The ratio of left ventricle weight to body weight （LVW/BW） was measured to reflect myocardial hypertrophy.The caudal arterial pressure was measured by tail-cuff.Protein expression of p-ERK in myocardial tissue was detected by Western blotting,BNP mRNA in myocardial tissue was examined by RT-PCR,and protein expression of plasma BNP was detected by ELISA.RESULTS：1.Benazepril and hydralazine lowered the blood pressure after 10 weeks treatment (P<0.01).2.The ratio of LVW/BW in SHR benazepril group was significantly lower than that in SHR hydralazine group and SHR sham group (P<0.01),and similar to that in WKY group (P>0.05).3.The protein expression of p-ERK in myocardial tissue in SHR benazepril group was significantly lower than that in SHR hydralazine group and SHR sham group (P<0.01),and similar to that in WKY group (P>0.05).There was no significant difference of p-ERK expression between SHR hydralazine group and SHR sham group (P>0.05).4.The levels of plasma BNP and BNP mRNA in myocardial tissue in SHR benazepril group were significantly lower than that in SHR hydralazine group and SHR sham group (P<0.01),and similar to that in WKY group (P>0.05).There was no significant difference of plasma BNP and BNP mRNA in myocardial tissue between SHR hydralazine group and SHR sham group (P>0.05).CONCLUSIONS：Benazepril inhibited ERK activation,resulting in regression of myocardial hypertrophy and accompanied by the reduction of BNP level.However,in spite of the effect of lowering blood pressure,hydralazine did not prevent or regress cardiac hypertrophy and did not decrease the level of p-ERK and BNP in SHR.BNP level might serve as a therapeutic index for reversal of myocardial hypertrophy.     

Establishment of miniature swine myocardial infarction model by percutaneous balloon occlusion method

AIM: To study the feasibility of establishing miniature swine myocardial infarction(MI)model by percutaneous balloon occlusion method, and to investigate the effect of autologous mesenchymal stem cells(MSCs)transplantation on treatment of acute MI. METHODS: Twenty Tibet miniatrue swine were included in the study and randomly divided into 2 groups. After anaesthesia, 2.5 mm×15.0 mm percutaneous transluminal coronary angioplasty balloon was positioned in the middle and distal-LAD by percutaneous femoral puncture in the right inguinal in the animals. The LAD flow was occluded for 90 min. The electrocardiogram(ECG), echocardiography(UCG), single photon emission computed tomography(SPECT), magnetic resonance imaging(MRI)and histopathology were examined. MSCs suspensions were injected to the infarct myocardium through infarct-related coronary artery by OTW balloon in MSCs transplantation group, while the control swine were injected with normal saline. Left ventricular ejection fraction(LVEF), end diastolic volume(EDV), end systolic volume(ESV)and fractional shortening(FS)were observed. RESULTS: The coronary angiography was successfully performed in all 20 swine. Ten swine survived during the process of balloon occlusion, 10 dead and the success rate reached 50%. After MI, LVEF and FS significantly decreased, while EDV and ESV significantly increased as compared to the values of pre-modeling(P<0.05). Eight weeks after transplantation, the parameters observed above improved in MSCs group as compared to those of pre-transplantation(P<0.05,P<0.01). However, no significant difference of the parameters in saline group between pre-transplantation and after-transplantation was observed. Compared to saline group, LVEF and FS significantly increased in MSCs group(P<0.05,P<0.01)with significant decreases in EDV and ESV(P<0.05,P<0.01). CONCLUSION: The miniature swine MI model is successfully established by percutaneous balloon occlusion method. Stem cells transplantation prevents the occurrence of ventricular fibrillation and obviously improves the myocardial functions.     

Effects of urocortin-I preconditioning on myocardial mitochondrial respiratory function and enzyme activity in rats

ATM: To investigate the influence of urocortin-I (Ucn I) preconditioning on the myocardial mitochondrial respiratory function and enzyme activity in the rats with ischemia reperfusion, and to observe the changes of ATP content in the myocardial cells. METHODS: (1) The healthy male Sprague-Dawley rats were randomly divided into 4 groups:normal group (Nor group), ischemia reperfusion group (IR group), Ucn I preconditioning group (Ucn I group), 5-hydroxy acid (5-HD)+Ucn I group. Langendorff perfusion was used to establish the in vitro model of cardiac ischemia reperfusion. At the end of the balance (T₁), before ischemia (T₂) and at the end of the reperfusion (T₃) respectively, the myocardial mitochondria was extracted, the mitochondrial respiratory function and respiratory enzyme activity in each group were determined. (2) The method of MPA isolated heart perfusion was used to isolate myocardial cells of the adult rats. After cultured for 24 h, myocardial cells were divided into 4 groups:Nor group, hypoxia/reoxygenation group (I/R group), Ucn I group, 5-HD+Ucn I group. Hypoxia/reoxygenation model of myocardial cells was established. At the end of reoxygenation, the changes of myocardial ATP content were measured by high performance liquid chromatography.RESULTS: (1) Compared with T₁, T₂ time points, the respiratory function (state 3 respiratory rate, respiratory control rate) and NADH oxidase, succinate oxidase and cytochrome C oxidase activities at T₃ time point were significantly decreased (P<0.05) in all groups except Nor group. At T₃ time point, the myocardial mitochondrial respiratory function and respiratory enzyme activity in Ucn I group were superior to 5-HD+Ucn I group and IR group (P<0.05), but was inferior to Nor group (P<0.05). At T₃ time point, the respiratory function of myocardial mitochondria and respiratory enzyme activities (NADH oxidase, succinate oxidase) in 5-HD+Ucn I group were better than those in IR group (P<0.05), but no statistical difference of the cytochrome C oxidase activity between the 2 groups was observed. The respiratory function and 3 kinds of respiratory enzyme activities at T₁, T₂ time points had no statistical change. (2) At the end of the reoxygenation, the myocardial ATP content in Nor group was higher than that in other groups (P<0.01). The myocardial ATP contents in I/R group and 5-HD+Ucn I group were lower than that in Ucn I group (P<0.05). In additon, 5-HD+Ucn I group was higher ATP content compared with I/R group (P<0.05). CONCLUSION: Ucn I preconditioning attenuates the ischemia/reperfusion induced damages of myocardial mitochondrial respiratory function and respiratory enzyme activity, thus ensuring the myocardial ATP contents under the condition of hypoxia/reoxygenation.     

Involvement of Sirt1 in mouse myocardial hypertrophy induced by isorenin

AIM: To investigate the expression and function of sirtuin 1 (Sirt1), one of the class III histone deacetylases, in hypertrophied mouse myocardium induced by isorenin (ISO). METHODS: The Kunming mice were randomly divided into 3 groups: control group, ISO group and ISO+nicotinamide (NAM) group. The myocardial hypertrophy was induced by dorsal subcutaneous injection of isorenin. Nicotinamide, an inhibitor against Sirt1, was given by peritoneal injection. Heart weight index (heart weight/body weight), hematoxylin and eosin staining, transmission electron microscope and mRNA expression of brain natriuretic peptide (BNP) were observed to identify the myocardial hypertrophy. The expression of Sirt1 in mRNA and protein levels was detected by RT-PCR and Western blotting, respectively. RESULTS: Compared to control group, the results of heart weight index, hematoxylin and eosin staining, the observation of transmission electron microscopy and mRNA expression of BNP showed that the mouse myocardial hypertrophy was induced by isorenin successfully. The Sirt1 expression was increased in hypertrophy model group (P<0.01 vs control group). Treatment with nicotinamide inhibited the cardiac hypertrophy induced by ISO (P<0.05 vs ISO group) and decreased the expression of Sirt1 (P<0.01 vs ISO group). CONCLUSION: Activation of Sirt1 might be involved in the process of myocardial hypertrophy stimulated by isorenin in mice.     

Cardiomyocyte apoptosis and related gene expression after acute myocardial infarction in rats

AIM: To investigate cardiomyocyte apoptosis and the expression of caspase-3, Bcl-2 and Bax after acute myocardial infarction (AMI) in rats.METHODS: AMI model was established with the ligation of left coronary artery in 78 randomly selected female SD rats.Twenty-four hours after operation, 43 survivors were randomly divided into 48-hour and 4-week two groups according to the time points: MI 48 h (n=11) and MI4 weeks (n=13) groups, sham-operated rats (S, n=27) were also randomly selected and reassigned to S48 h (n=10) and S4 weeks (n=10) groups.Cardiomyocyte apoptosis was detected with in situ terminal deoxynucleotidyl transferase (TdT)-dUTP nick-end labeling (TUNEL staining) and DNA gel electrophoresis.Caspase-3, Bcl-2 expression and Bax expression were detected with immunohistochemistry and Western blotting analysis.RESULTS: Compared with sham-operated group, after AMI, systolic, diastolic, and mean arterial blood pressures (SBP, DBP, MAP), left ventricular systolic pressure (LVSP) and the maximum change rate of left ventricular pressure rise and fall (±dp/dt) were significantly decreased (P<0.05, P＜0.01), while left ventricular end diastolic pressure (LVEDP) was significantly increased (P<0.05) in MI 48 h group.All the above indices in MI 4 weeks group had the same change as that in MI48h group, with the LVEDP significantly higher (P<0.01), except for a non-significantly change in SBP, DBP and MAP (all P>0.05).In both MI 48 h and MI 4 weeks groups, myocyte apoptotic index was significantly increased in the infracted/scar, border and non-infarcted areas (P<0.05,P＜0.01) with caspase-3 and Bax expressions increased significantly (P<0.05, P＜0.01) in myocytes of the above three areas and Bcl-2 expression increased only in myocytes of the infracted area in MI 48 h group.Western blotting indicated that Bcl-2/Bax ratio was also decreased in MI 48 h subgroup.CONCLUSIONS: After AMI in rats, cardiomyocyte apoptosis happened in the infarction/scar, border and non-infarcted areas, with caspase-3 and Bax expression in myocytes increased, and with Bcl-2 expression increased in myocytes of infracted area and Bcl-2/Bax ratio decreased only early after AMI.     

Effect of fluvastatin on left ventricular remodeling and caspase-3 expression after myocardial infarction in rats

AIM: To observe the effect of fluvastatin (FV) on left ventricular remodeling and expression of caspase-3 after myocardial infarction (MI) in rats. METHODS: Rats were divided into 4 groups: group Ⅰ (sham), group Ⅱ (sham+FV), group Ⅲ (MI) and group Ⅳ (MI+FV). group Ⅱ and Ⅳ were treated with FV (10 mg·kg^-1·d^-1) for 4 weeks. The left ventricular structure, echocardiography and hydroxyproline were observed. The expression of caspase-3 was measured by immunohistochemistry and RT-PCR. RESULTS: Compared with MI group, there was a improvement of ultrastructure and index of left ventricular remodeling, and decrease in hydroxyproline in MI+FV group (all P<0.05). The number of caspase-3 positive cells also decreased in MI+FV group, and RT-PCR showed the level of caspase-3 mRNA expression was lower than that in MI group (P<0.05). CONCLUSION: Fluvastatin improves left ventricular remodeling after myocardial infarction, decreases the expression of caspase-3 and inhibits apoptosis.     

Zacopride attenuates pressure overload-induced ventricular remodeling in rats

AIM:To investigate the protective effect of zacopride (ZAC) on the pressure-overload left ventricular remodeling in the rats induced by coarctation of abdominal aorta. METHODS:Male Sprague-Dawley (SD) rats with pressure overload were induced by the coarctation of abdominal aorta. The model rats were intraperitoneally administered with ZAC, chloroquine (Chlor), and zacopride+chlorquine (ZAC+Chlor). The study duration was 8 weeks. The cardiac structure and function were assessed by echocardiography. The heart weight/body weight (HW/BW) ratio and the left ventricular weight/body weight (LVW/BW) ratio were calculated. The changes of structure and shape in myocardial tissue were observed with HE staining. The ultrastructure of the myocytes was observed under transmission electron microscope. The inward rectifier potassium channel (I_K1) protein expression was determined by Western blot. The mRNA expression of Kir2.1 was detected by RT-PCR. RESULTS:Compared with vehicle group, ZAC improved cardiac function, as indicated by the decreased left ventricular end-diastolic dimension (LVEDD) and left ventricular end systolic dimension (LVESD) (P<0.05), and the increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (P<0.01). The HW/BW and LVW/BW ratios were significantly decreased, and the cross-sectional area of the cardiomyocytes was significantly less in ZAC group than that in vehicle group (P<0.01). The ultrastructure of the myocytes was significantly improved. Chlor blocked the protective effect of zacopride on the pressure-overload left ventricular remodeling. The protein level of I_K1 and mRNA expression of Kir2.1 in the cardiac tissues in ZAC group were significantly increased compared with vehicle group (P<0.01). CONCLUSION:I_K1 agonist ZAC significantly attenuates pressure overload-induced ventricular remodeling in rats.