Ratio Of Heart Weight Research Articles

Studies of Body Weight/Heart Weight [C/S] Ratio in Treated and Untreated Experimental Pulmonary Barotrauma

Abstract The prerequisite of development of pulmonary barotrauma [PB] is retention of the breathing mix in the lungs during a sudden decrease in external pressure or its administration into the airways under increased pressure or in a volume exceeding the maximum lung capacity. In such cases, the pulmonary parenchyma ruptures and air enters both the pleural cavity and/or the lumen of ruptured blood vessels located in the alveolar septa. The result is permanent disruption of the pulmonary parenchyma. The aim of the study was to assess the influence of post-PB lesions on the heart muscle and the importance of hyperbaric treatment on the exacerbation of such lesions in the heart. The hearts of 35 rabbits were used in the study. In animals of the experimental group, PB was induced in the pressure chamber using the proprietary method described in previous publications. Part of the animals in this group were treated with air hyperbaria. The comparison group consisted of animals, which did not undergo PB during a simulated dive. All animals were weighed, observed for four weeks and then put to death following the experiment. In autopsy, among others, whole hearts were collected and weighed after fixation. Subsequently, the C/S ratio, i.e. the body to heart weight ratio, was calculated. The measurement results were subject to statistical analysis. A statistically significant increase in the C/S ratio was found, indicating an increase in the share of heart weight in the total body weight in the group of animals with PB not treated with air hyperbaria as compared to the control group.

P1624Blockade of protease activated receptor-1 signaling attenuates cardiac hypertrophy and fibrosis in renin-overexpressing hypertensive mice

Abstract Introduction Recent evidences have demonstrated that coagulation pathway is involved in cardiovascular remodeling induced by renin-angiotensin system (RAS), which finally leads to heart failure. Protease activated receptor-1 (PAR-1) is widely expressed in the vasculature and the heart, and plays important roles in pro-inflammatory process in the cardiovascular system. Recently, we demonstrated that the activity of factor Xa (FXa), which functions not only as a coagulation factor but as an agonist for PAR-1, was enhanced in renin-overexpressing hypertensive mice (Ren-Tg). Purpose The purpose of this study was to investigate whether inhibition of PAR-1 signaling has protective effects on the progression of heart failure induced by chronic RAS activation in Ren-Tg. Methods and results We treated 12–16 weeks-old male wild type mice (WT) and Ren-Tg with continuous subcutaneous infusion of PAR-1 antagonist SCH79797 (25mg/kg/day) or vehicle for 4 weeks. After treatment period, left ventricular (LV) wall thickness calculated as interventricular septum plus posterior wall thickness measured by echocardiography was greater in Ren-Tg than in WT (0.25±0.003 versus 0.18±0.002 mm), and SCH79797 attenuated the increase to 0.22±0.01 mm in Ren-Tg (both p<0.05, respectively). The ratio of heart weight to body weight was greater in Ren-Tg than in WT (6.1±0.4 versus 4.6±0.7 mg/g), and SCH79797 attenuated the increase to 5.2±0.1 mg/g (both p<0.05). The area of cardiac fibrosis evaluated by Masson-trichrome staining was greater in Ren-Tg than in WT (2.6±0.2 versus 1.4±0.3%), and SCH79797 attenuated it to 1.6±0.3% in Ren-Tg (both p<0.05). Cardiac mRNA expressions of tumor necrosis factor-α, transforming growth factor-β1, and β-myosin heavy chain were all greater in Ren-Tg than in WT, and SCH79797 attenuated the increases in Ren-Tg (all p<0.05). Conclusions Inhibition of PAR-1 signaling attenuates cardiac hypertrophy and fibrosis in Ren-Tg via inhibition of inflammatory cytokines production. These results support the involvement of PAR signaling in the development of heart failure induced by RAS, and may provide novel therapeutic insights for the treatment of hypertensive heart failure.

Effects of hypericum perforatum extract on the immunogenic myocardial fibrosis in mice and its mechanism

To study the effects of hypericum perforatum extract (HPE) on myocardial fibrosis of the experimental autoimmune myocarditis (MFEAM) in mice. MFEAM model of immunosuppressive mouse was established by porcine cardiac myosin. The mice in the MFEAM model group were randomly divided into: MFEAM model group (n=14), HPE 100 mg/kg group (n=13), HPE 40 mg/kg group (n=13) and captopril (Cap) 50 mg/kg control group (n=13). Drug in each group was dissolved in 0.4 ml of physiological saline each time, and administered by intragastric administration twice a day for 60 days; the normal control group (n=10) and the MFEAM model group was given normal saline at the same volume and course of treatment as above described way. The ratio of heart weight and spleen weight to body weight were calculated, the general conditions of mice were observed. The contents of procollagen I N-terminal peptide(PINP), procollagen III N-terminal peptide (PIIINP) and TGF-β1 in serum of mice were detected. The degree of myocardial fibrosis (MF) and collagen volume fraction (CVF)were measured under the microscope by Masson staining. The protein expression of TGF-β1 was detected by Western blot. The serum levels of TGF-β1, PINP and PIIINP in the model group were higher than those in the normal group significantly (P＜0.05 or P＜0.01). Meanwhile , the levels of MF and CVF were increased, and the protein expression of myocardial TGF-β1 was up-regulated. However, the serum levels of PINP, PIIINP and TGF-β1 in HPE40 mg/kg, HPE 100 mg/kg and Cap treated-groups were decreased significantly. HPE and Cap could improve or decrease the MF level and CVF of the MFEAM model mice, and down-regulated the expression of TGF-β1 protein in different degrees. Compared with the MFEAM model group, the difference was significant (P＜0.05 or P＜0.01). HPE has a therapeutic effect on MF, which may be related to its reduction of collagen deposition and inhibition of TGF-β1 signaling pathway.

Abstract P140: The Role of Pumpkin Flesh in Reducing Mean Arterial Pressure in a Genetic Rat Model for Hypertension

Objective: ACE inhibitors (ACEi) remain the most commonly prescribed medication; lowering blood pressure, increasing bradykinin levels and decreasing cardiac hypertrophy. A demographic-specific bradykinin sensitivity renders African-American patients on ACEi with elevated risk of angioedema. This increased risk has led a search for natural ACEi from functional foods or natural resources, one of which being pumpkin. In Sprague Dawley rats with L-NAME induced hypertension, pumpkin seed oil significantly reduced systolic blood pressure. As a promising source of β-carotene, phenolics and a beneficial fatty acid profile, this study aimed to examine the potential of pumpkin flesh and seed in regulating blood pressure management in a genetic model for hypertension. Methods: Spontaneously Hypertensive Rats (SHR) and Wistar-Kyoto rats (WKY), aged 6 weeks, were fed one of the following diets for 8 weeks: 4% pumpkin flesh (PF), 4% pumpkin seed (PS), control diet (Ctrl), and an SHR-Captopril treated group (Cap). Non-invasive blood pressure monitoring was employed for weekly measuring of mean arterial pressure (MAP). Upon 8 weeks of treatment, animals were euthanized for structural analysis of cardiac hypertrophy (Heart weight to tibial length ratio). Left ventricular mRNA expression of ANP and BNP was quantified using quantitative RT-PCR techniques. Results: At 8 weeks, MAP in SHR-PF and SHR-PS groups was significantly reduced by 23% (p=0.0135) and 27% (p=0.0018) respectively, compared to SHR-Ctrl. Heart weight ratios showed no significant difference between SHR-Ctrl and SHR-PF or SHR-PS, while SHR-Cap heart weight significantly decreased by 15% (p=0.0028) in comparison to SHR-Ctrl. Additionally, increased mRNA expression of both ANP and BNP was observed in the SHR-PF group in comparison to WKY-Ctrl. Conclusions: The reduction of blood pressure observed in response to pumpkin-based diets suggests an alternative pathway of blood pressure control independent of ACE inhibition. This potential new pathway elucidates the health promoting capabilities of pumpkin as an alternative blood pressure treatment for all demographics.

Abstract 265: Thyrotoxic Pericarditis in a Mouse Model of Graves' Disease

Objectives: To investigate the cardiac pathological changes of hyperthyroid heart disease in mice. Methods: A transient model for human Graves’ disease was successfully established in mice using three immunizations with recombinant adenovirus expressing the extracellular A subunit of the human thyrotropin receptor (Ad-TSHR). We induced the hyperthyroid heart mouse model by prolonging the regular injections of Graves’ disease mouse model to nine months. Heart tissues were weighed and stained with hematoxylin-eosin (HE) and MASSON Trichrome staining to check and define the pathological changes. Results: We observed that mice in the Graves’ disease hyperthyroid heart group (GH) showed enhanced cardiac hypertrophy and fibrosis. The HW/BW (ratio of heart weight to body weight) of GH group (5.528±0.4014, n=6) was significantly higher than control group (4.343±0.03913, n=6) (P<0.05). In GH group, 2 of 6 hearts showed typical cor villosum (Image 1). The MASSON Trichrome staining exhibited aggravated cardiac fibrosis on the surface of the heart in GH group. Conclusions: Pericarditis as a complication of thyrotoxicosis has been reported several times in clinical work. This is the first reported case of pericarditis in the mouse model of Graves’ disease, suggesting a possible link between pericarditis and thyrotoxicosis.

Total Astragalus saponins attenuates CVB3-induced viral myocarditis through inhibiting expression of tumor necrosis factor α and Fas ligand.

To investigate the therapeutic effect of total Astragalus saponins (AST) against viral myocarditis in animal and cell models. Primary myocardiocytes (PMCs) were stimulated by the coxsackie B (CVB) 3 virus to prepare the cell model of viral myocarditis. Cell viability, apoptosis and the mRNA expression of C-Myc, tumor necrosis factor (TNF)-α and Fas were detected to evaluate the protective effects of AST on CVB3-induced PMC damage. AST could significantly increase survival and decrease the ratio of heart weight: body weight (P<0.05). The level of myocardial fibrosis in the AST group was significantly lower than that in the CVB3 group. Compared with the CVB3 group, the ejection fraction was increased significantly in the AST group. Levels of lactate dehydrogenase and creatine kinase-MB in the peripheral blood of the AST group were significantly lower than those in the control group. In vitro, AST could significantly decrease CVB3-induced PMC apoptosis. Expression of C-Myc, TNF-α, Fas in the AST group was significantly lower than that in the CVB3 group. It is demonstrated that AST was protective against CVB3-induced viral myocarditis, which may be associated with a decrease in CVB3-induced apoptosis and down-regulation of expression of C-Myc, TNF-α and Fas.

Klotho inhibits angiotensin II-induced cardiac hypertrophy, fibrosis, and dysfunction in mice through suppression of transforming growth factor-β1 signaling pathway

Recent studies have revealed critical roles of transforming growth factor-β1 (TGF-β1) and microRNA-132 (miR-132), a downstream mediator of TGF-β1, in the pathogenesis of cardiac remodeling. In this study, we tested whether the antiaging protein klotho modifies angiotensin II (Ang II)-induced cardiac remodeling through regulating TGF-β1-miR-132 axis. We found that both klotho and the TGF-β1 inhibitor LY364947 significantly inhibited cardiac hypertrophy, fibrosis, and dysfunction in Ang II-infused mice, as evidenced by the ratios of heart weight to body weight (HW/BW), heart weight to tibial length (HW/TL), cardiomyocyte cross-sectional area, fibrotic area, and expression of prohypertrophic genes (ANP, β-MHC) and fibrotic marker genes (α-SMA, collagen I), echocardiographic parameters. Meanwhile, klotho also significantly inhibited Ang II-induced protein expression of TGF-β1 and phosphorylated Smad2/3 in the heart tissues and cultured cardiomyocytes and cardiac fibroblasts. In vitro experiments demonstrated that Ang II-induced cardiomyocyte hypertrophy and proliferation and activation of cardiac fibroblasts were markedly inhibited by klotho, LY364947 or the miR-132 inhibitor anti-miR-132. Both klotho and the TGF-β1 inhibitor LY364947 downregulated the miR-132 expression. Additionally, klotho decreased Ang II-induced protein expressions of cardiac fibroblast growth factor (FGF)23 in vivo and in vitro. The decreased protein levels of klotho in serum and renal tissues of Ang II-infused mice were elevated by klotho. Klotho downregulated the protein levels of TGF-β1 in renal tissues of Ang II-infused mice. In conclusion, our results suggest that klotho prevents Ang II-induced cardiac remodeling and dysfunction through modifying the TGF-β1-miR-132 axis, providing an experimental basis for clinical treatment on cardiac remodeling.

Loss of microRNA-27a induces cardiac dysfunction through activating FoxO1.

To elucidate how microRNA-27a and FoxO1 regulate cardiac dysfunction in mice. Expression levels of ANP, BNP, β-MHC, α-SMA, Fn1, and Periostin in myocardial tissues of 2-month-old and 8-month-old microRNA-27a-KO mice and age-matched wild-type mice were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. Dual-luciferase reporter gene assay was conducted in H9C2 cells to verify the binding condition between microRNA-27a and FoxO1. By transfection of microRNA-27a mimics or inhibitor, FoxO1 expression in H9C2 cells was determined at the mRNA and protein levels. HW/BW [ratio of heart weight (mg) and body weight (mg)], HW/TL [ratio of heart weight (mg) and tibial length (mm)], LVPWDT [left ventricular posterior wall diastolic thickness (mm)], LVEDD [left ventricular end-diastolic dimension (mm)], and FS (fractional shortening) in mice treated with or without FoxO1 inhibitor AS1842856 were accessed through echocardiography. MicroRNA-27a-KO mice had larger LVEDD, HW/BW, and HW/TL, but lower FS and LVPWDT than those of age-matched wild-type mice. Besides, higher levels of ANP, BNP, β-MHC, α-SMA, Fn1, and Periostin were observed in myocardial tissues of microRNA-27a-KO mice compared with those of age-matched wild-type mice. Dual-luciferase reporter gene assay revealed lower luciferase activity in H9C2 cells co-transfected with microRNA-27a mimics and wild-type FoxO1 than that of controls. The expression level of FoxO1 was negatively regulated by microRNA-27a in H9C2 cells at the mRNA and protein levels. After AS1842856 injection, HW/BW, HW/TL, and LVEDD in microRNA-27a-KO mice markedly decreased, whereas FS and LVPWDT elevated. By comparison, AS1842856 injection did not influence cardiac development in wild-type mice. MicroRNA-27a knockout could induce cardiac dysfunction in mice through upregulating FoxO1 expression.

Acupuncture at PC6 prevents cardiac hypertrophy in isoproterenol-treated mice.

To investigate the effect of acupuncture at PC6 on cardiac hypertrophy in isoproterenol (ISO)-treated mice. 48 male C57BL/6 mice underwent subcutaneous injection of ISO for 14 days and were randomly divided into four groups (n=12 each) that remained untreated (ISO group), received verum manual acupuncture (MA) treatment at PC6 (ISO+MA(PC6) group), sham MA at location on the tail not corresponding to any traditional acupuncture point (ISO+MA(tail) group), or propranolol (ISO+PR group). An additional 12 mice were given an injection of phosphate-buffered saline (PBS) and formed a healthy control (Normal) group. After performing echocardiography and measuring the ratio of heart weight (HW)/tibia length (TL) at 14 days, all mice were euthanased. Morphological examination was performed following haematoxylin and eosin and Masson's staining of heart tissues. Ultrastructural changes were observed by electron microscopy. Cardiac protein expression of atrial natriuretic peptide (ANP) and tumour necrosis factor α (TNFα) were measured by immunohistochemical (IHC) staining and Western blotting. Compared with the untreated model group, acupuncture at PC6 lowered the heart rate, reduced the ratio of HW/TL, improved the left ventricular (LV) anterior wall thickness (LVAWd), LV end-diastolic anterior wall thickness (LVAWs), LV end-systolic posterior wall thickness (LVPWd), LV end-diastolic posterior wall thickness (LVPWs), and fractional shortening (FS) as observed by echocardiography (ISO+MA(PC6) vs. ISO groups: P<0.05). Moreover, evidence from morphological studies demonstrated that acupuncture at PC6 inhibited myocardial hypertrophy and collagen deposition, and normalised the ultrastructural changes. In addition, ANP and TNFα expression were attenuated in the verum acupuncture group compared with the untreated model group (ISO+MA(PC6) vs. ISO groups: P<0.05). The results demonstrated that acupuncture at PC6 attenuates sympathetic overactivity. Additionally, it may improve cardiac performance by reversing adverse cardiac remodelling. Acupuncture has potential as a treatment for sympathetic hypertension.

Magnesium supplementation improves diabetic mitochondrial and cardiac diastolic function.

In heart failure and type 2 diabetes mellitus (DM), the majority of patients have hypomagnesemia, and magnesium (Mg) supplementation has improved cardiac function and insulin resistance. Recently, we have shown that DM can cause cardiac diastolic dysfunction (DD). Therefore, we hypothesized that Mg supplementation would improve diastolic function in DM. High-fat diet-induced diabetic mouse hearts showed increased cardiac DD and hypertrophy. Mice with DM showed a significantly increased E/e' ratio (the ratio of transmitral Doppler early filling velocity [E] to tissue Doppler early diastolic mitral annular velocity [e']) in the echocardiogram, left ventricular end diastolic volume (LVEDV), incidence of DD, left ventricular posterior wall thickness in diastole (PWTd), and ratio of heart weight to tibia length (HW/TL) when compared with controls. DM mice also had hypomagnesemia. Ventricular cardiomyocytes isolated from DM mice exhibited decreased mitochondrial ATP production, a 1.7- ± 0.2-fold increase of mitochondrial ROS, depolarization of the mitochondrial membrane potential, and mitochondrial Ca2+ overload. Dietary Mg administration (50 mg/ml in the drinking water) for 6 weeks increased plasma Mg concentration and improved cardiac function. At the cellular level, Mg improved mitochondrial function with increased ATP, decreased mitochondrial ROS and Ca2+ overload, and repolarized mitochondrial membrane potential. In conclusion, Mg supplementation improved mitochondrial function, reduced oxidative stress, and prevented DD in DM.

Maternal high-fat diet alters angiotensin II receptors and causes changes in fetal and neonatal rats†.

Maternal high-fat diet (HFD) during pregnancy is linked to cardiovascular diseases in postnatal life. The current study tested the hypothesis that maternal HFD causes myocardial changes through angiotensin II receptor (AGTR) expression modulation in fetal and neonatal rat hearts. The control group of pregnant rats was fed a normal diet and the treatment group of pregnant rats was on a HFD (60% kcal fat). Hearts were isolated from embryonic day 21 fetuses (E21) and postnatal day 7 pups (PD7). Maternal HFD decreased the body weight of the offspring in both E21 and PD7. The ratio of heart weight to body weight was increased in E21, but not PD7, when compared to the control group. Transmission electron microscopy revealed disorganized myofibrils and effacement of mitochondria cristae in the treatment group. Maternal HFD decreased S-phase and increased G1-phase of the cellular cycle for fetal and neonatal cardiac cells. Molecular markers of cardiac hypertrophy, such as Nppa and Myh7, were found to be increased in the treatment group. There was an associated increase in Agtr2 mRNA and protein, whereas Agtr1a mRNA and AGTR1 protein were decreased in HFD fetal and neonatal hearts. Furthermore, maternal HFD decreased glucocorticoid receptors (GRs) binding to glucocorticoid response elements at the Agtr1a and Agtr2 promoter, which correlated with downregulation of GR in fetal and neonatal hearts. These findings suggest that maternal HFD may promote premature termination of fetal and neonatal cardiomyocyte proliferation and compensatory hypertrophy through intrauterine modulation of AGTR1 and AGTR2 expression via GR dependent mechanism.

Rapid Communication: A comparison of cardiac lesions and heart weights from market pigs that did and did not die during transport to one Ontario abattoir.

In-transit losses of market hogs represent a small proportion of all market-weight pigs shipped in a year. This suggests that individual pig factors may be a significant cause of in-transit losses along with more traditionally considered environmental and transport factors. An investigation was performed to determine whether cardiac pathology and heart weights were associated with pigs that did or did not die during transport to an abattoir. The hearts from 70 pigs that died in-transit to one Ontario abattoir and 388 pigs that arrived alive were collected and examined. Hearts from pigs that died during transport demonstrated greater frequencies of cardiac lesions (P < 0.05). These included hypertrophy of ventricle walls (Left: 97% vs. 64%; Right: 86% vs. 57%), dilation of ventricle chambers (Left: 79% vs. 0.5%; Right: 100% vs. 5%), and dilation of the pulmonary artery and aorta (59% vs. 1.5%). Total heart weight to body weight ratios were increased (3.6 vs. 3.3 g/kg) and left ventricle plus septum weight over right ventricle weight ratio was decreased in pigs that died during transport over non–in-transit loss pigs (2.5 vs. 2.8; P < 0.05). This may indicate reduced cardiac function in hogs that died during transport. Pigs with reduced cardiac function would have exercise intolerance and be more susceptible to death during transport due to the increased cardiac workload required during sorting, loading, and transport of the pigs to the abattoir. Further research to quantify cardiac function in pigs with cardiac lesions or abnormal heart weight ratios is warranted.

Downregulated microRNA-224 aggravates vulnerable atherosclerotic plaques and vascular remodeling in acute coronary syndrome through activation of the TGF-β/Smad pathway.

Recent studies have shown that circulating microRNAs (miRNA) play a critical role in diagnosing acute coronary syndrome (ACS). This study aims to investigate the effect of miR-224 on atherosclerotic plaques forming and vascular remodeling in ACS and its relationship with TGF-β/Smad pathway. Myocardial infarction (MI) rat model was established and lentivirus vector of miR-224 inhibitor was prepared for investigating the effect of downregulated miR-224 on the contents of nitric oxide (NO) and endothelin-1 (ET-1), blood lipid levels and inflammatory factor levels in serum as well as the TGF-β/Smad pathway. The rats suffering from MI had decreased survival rates and exhibited reduced levels of NO, high-density lipoprotein cholesterol, and lumen diameter, and Smad7 messenger RNA (mRNA) and protein expression; while had significantly increased ratio of heart weight or body weight, levels of ET-1, inflammatory factors, blood lipid indexes, vascular remodeling indexes, collagen volume fraction, vulnerable atherosclerotic plaque area, VCAM-1 and MMP-2 protein expression, TGF-β, Smad2, Smad3, and Smad4 mRNA and protein expression. After inhibiting the TGF-β/Smad pathway, the rats suffering from MI showed notably opposite trend. In conclusion, downregulation of miR-224 expression promotes the formation of vulnerable atherosclerotic plaques and vascular remodeling in ACS through activation of the TGF-β/Smad pathway. Therefore, this study provides a new therapeutic target for ACS.

Abstract 409: Ablation of Endothelial Sirt3 Exacerbates Pressure Overload-induced Heart Failure

Cardiovascular disease leads to cardiac dysfunction and eventually heart failure which is the leading cause of death in the United States. Our previous data indicates that global ablation of SIRT3 exacerbates acute injury after myocardial ischemia and that deletion of endothelial SIRT3 (SIRT3 ECKO) leads to development of cardiac dysfunction. In the present study, we demonstrates that loss of SIRT3 in endothelial cells results in upregulation of TIGAR, but downregulation of HIF-1α, HIF-2α, glucose transport GLUT-1, glycolytic enzymes PFK-1, and PFKFB3. To further investigate the role of endothelial SIRT3 on cardiac hypertrophy and heart failure during chronic stress, SIRT3 ECKO and their control SIRT3 LoxP mice were subjected to pressure overload-induced by transverse aortic constriction (TAC) for 7 weeks. Our results demonstrate that the ratio of heart weight to tibia length was increased in both strains of mice, in which SIRT3 ECKO + TAC mice exhibited more severe cardiac hypertrophy. In addition, systolic function was significantly decreased in SIRT3 ECKO + TAC mice compared to SIRT3 LoxP + TAC mice, as evidenced by lower EF and FS at week 7. In contrast, diastolic function decreased after TAC surgery in a similar manner in both strains of mice, despite that pre-existing diastolic dysfunction was found in SIRT3 ECKO mice. Similarly, there was no difference in coronary flow reserve between both strains of mice subjected to TAC. Moreover, pressure overload-induced upregulation of Apelin, Angiopoietin-1, GLUT-4, and ERK phosphorylation were decreased in SIRT3 ECKO + TAC mice, whereas the expression of pro-inflammatory molecules, ICAM-1 and VCAM-1, was significantly increased in SIRT3 ECKO + TAC mice. These results implicate a critical role of SIRT3 in regulating glucose uptake and glycolytic function in endothelial cells. Ablation of endothelial SIRT3 exacerbates pressure overload-induced cardiac hypertrophy and heart failure via impaired glucose metabolism and increased pro-inflammatory molecules.

The involvement of AQP1 in myocardial edema induced by pressure overload in mice.

To investigate the effect of aquaporin-1 (AQP1) on heart edema induced by transverse aortic constriction (TAC) in mice, and to explore whether inhibiting the expression of AQP1 could attenuate myocardial edema and improve cardiac function. The C57BL/6 mice were divided into four groups: (1) the sham group; 2) the sham + acetazolamide group: mice were orally gavaged with acetazolamide (20 mg/kg/day) after sham operation; (3) the TAC group: a mouse model of pressure overload induced by TAC for two weeks; (4) the TAC + acetazolamide group: mice were orally gavaged with acetazolamide (20 mg/kg/day) after TAC. Cardiac function was detected by echocardiography after 2 weeks' TAC. The ratio of heart weight to body weight (HW/BW) and myocardial water content were calculated. The mRNA and protein expressions of AQP1 were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot, respectively. Significant myocardial hypertrophy and dysfunction were found in TAC mice. The ratio of HW/BW, myocardial water content, and the mRNA and protein expression of AQP1 of the TAC group were markedly higher than those of the sham group. By contrast, acetazolamide administration reduced the ratio of HW/BW and myocardial water content, whereas improved cardiac dysfunction induced by TAC. Moreover, acetazolamide reduced the mRNA and protein expression of AQP1 in TAC mice. The expression of AQP1 was closely related to myocardial edema induced by TAC. The inhibition of AQP1 could reduce myocardial edema and improve cardiac dysfunction.

Prorenin independently causes hypertension and renal and cardiac fibrosis in cyp1a1-prorenin transgenic rats

Plasma prorenin is commonly elevated in diabetic patients and appears to predict the development of diabetic nephropathy. However, the pathological role of prorenin is unclear. In the present study, a transgenic, inducible, hepatic prorenin-overexpressing rat model was generated and the effect of prorenin in organ injury was examined. Four groups of rats (cyp1a1 prorenin transgenic male and female rats and non-transgenic littermates) were assigned to receive a diet containing 0.3% of the transgene inducer indole-3-carbinol (I3C) for 4 weeks. Plasma prorenin concentration was increased and mean arterial pressure (MAP) increased from 80 ± 18 to 138 ± 17 (mmHg), whereas renal prorenin/renin protein expression was unchanged, in transgenic rats fed with I3C diet. The intact prorenin, not renin, in plasma and urine samples was further observed by Western blot analysis. Importantly, transgenic rats with high levels of prorenin developed albuminuria, glomerular and tubulointerstitial fibrosis associated with increased expression of transforming growth factor β (TGFβ) 1 (TGFβ1), plasminogen activator inhibitor-1 (PAI-1), collagen, and fibronectin (FN). These rats also exhibited cardiac hypertrophy determined by echocardiography, with elevated ratio of heart weight to body weight (HW/BW). Cardiac collagen in interstitial and perivascular regions was prominent, accompanied by the increase in mRNA contents of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), β-myosin heavy chain (β-MHC), TGFβ1, PAI-1, and collagen in the heart tissue. Furthermore, renal protein levels of p-NF-κB-p65 and monocyte chemoattractant protein-1 (MCP-1), NAPDH oxidases, malondialdehyde (MDA) and 8-isoprostane (8-IP), p-ERK, p-β-catenin, and p-Akt were dramatically increased in prorenin overexpressing rats. These results indicate that prorenin, without being converted into renin, causes hypertension, renal and cardiac fibrosis via the induction of inflammation, oxidative stress and the ERK, β-catenin, and Akt-mediated signals.

Seven Years of Experiences of Preclinical Experiments of Xeno-Heart Transplantation of Pig to Non-Human Primate (Cynomolgus Monkey)

BackgroundThe absolute shortage of donors compared with patients requiring transplantation is currently an unsolved problem, and the only possible solution may be xenotransplantation. To establish a successful clinical trial, a preclinical study using nonhuman primates is essential. Starting in November 2011, our team initiated heterotopic abdominal heart xenotransplantation, the first in the Republic of Korea. We present here the initial 7-year results. MethodsA total of 22 xenotransplantation procedures have been performed since 2011. Single transgenic pig (alpha-galactosidase transferase knockout [GalT KO], n = 16), double transgenic pig (GalT KO + CD46, n = 3, and GalT KO + CD39, n = 2), and triple transgenic pig (GalT KO + CD46 + CD70, n = 1) models were used. Our baseline regimen of immunosuppressants comprised CD154 ab, rituximab, anti-thymocyte globulin, tacrolimus, mycophenolate mofetil, and steroids. ResultsThe mean graft survival was 16 ± 16.27 days, and the mean graft survival was significantly longer in cases performed since 2014 (7.5 ± 8.03 days vs 24.67 ± 17.50; P = .01). Although the donor heart ischemic time was decreased per annum, no correlations could be found between ischemic time and survival days of the graft. Double or triple genetic manipulated hearts exhibited significantly better survival (11.63 ± 11.29 days vs 30.83 ± 20.34 days; P = .03). When the ratio of heart weight (grams) to nonhuman primate weight (kilograms) was lower, the results tended to be better (P < .05). The rate of immediate postoperative bleeding (9%, n = 2) causing death was relatively high in the earlier period, but there have been no serious surgical complications affecting graft survival since 2013. ConclusionsInvestigation of effective and optimal target genes for each organ to further progression toward better results is important. In addition, the immunosuppressive regimen needs to be further studied and constantly refined.

Effect of Shikonin on Cardiac Wound Healing in Mice

Shikonin has been shown to have properties of improving open wound healing, but it is not known whether shikonin is effective in promoting cardiac repair after myocardial infarction (MI). We hypothesize that shikonin can improve wound healing in infarcted hearts. We created MI models by ligating the coronary artery in mice. We treated the infarcted mice with 5 mg/kg of shikonin a day through oral gavage, or the same volume of distilled water was given to the control group. After seven days of treatment, we documented the heart weight and body weight, and the hearts were photographed and the infarct size was measured. The ratio of heart weight to body weight was calculated.We found that the infarct area in treated mice was significantly smaller than in the control group. While there was no significant difference in body weight between the two groups, but the ratio of heart weight to body weight in the treated group was much smaller than in the control group. In conclusion, shikonin was effective in helping wound healing in the heart after MI. Future studies will be directed to further study how shikonin functions at the cellular level and whether it improves cardiac function.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effects of Supplemental Epigallocatechin Gallate in the Diet of Broilers Exposed to Fluoride Intoxication.

We evaluated the effects of dietary epigallocatechin gallate (EGCG) on the performance, biochemical parameters, and liver histopathology of fluoride-intoxicated broiler chickens. In total, 160 1-day-old male broiler chicks (Ross PM3 strain) were collected and assigned to four groups (40 animals each), with four replicates. The control group received a basal diet; the F group received 800mg/kg fluoride; the EGCG group received 400mg/kg EGCG; and the EGCG+F group received 400mg/kg EGCG and 800mg/kg fluoride. The live weight (LW) of F-treated chicks was significantly lower than that of the controls. In the F-treated groups, feed intake (FI) and LW values were lower, but the feed conversion ratio (FCR) was higher than those of the controls. The ratio of heart weight to LW was found to be the highest in the F-treated groups. Alkaline phosphatase (ALP), aspartate aminotransferase (AST), and total oxidant status (TOS) levels in the F-treated groups were significantly higher, whereas the increase in total cholesterol levels was insignificant than those in the control group. In the EGCG+F group, AST, total cholesterol, and TOS levels decreased to a level comparable to those in the control group. Histopathological evaluation revealed that there were mild changes in the portal region in the EGCG+F group; additionally, there was an improvement in liver morphology in the EGCG+F group compared to that in the F group. Thus, EGCG has potent antioxidant and regenerative effects that can ameliorate the detrimental effects of fluoride toxicity on blood parameters and the liver.

Gambogic acid exerts cardioprotective effects in a rat model of acute myocardial infarction through inhibition of inflammation, iNOS and NF-κB/p38 pathway.

Gamboge, the dried resin secreted by Garcinia maingayii (gambogic tree), was previously demonstrated to exert anti-inflammatory effects. The present study examined the effects of gambogic acid, the major active constituent of gamboge, on myocardial infarction (MI) and inflammation in a rat model and explored the possible underlying mechanisms. The results demonstrated that gambogic acid inhibited the ratio of heart weight to body weight and myocardial damage (via lactate dehydrogenase and cardiac troponin T) in rats with MI. Gambogic acid suppressed the activation of interleukin (IL)-6 and tumor necrosis factor-α, and increased IL-10 levels in MI rats. Furthermore, gambogic acid reduced inducible nitric oxide synthase (iNOS), matrix metalloproteinase (MMP)-2, MMP-9, intercellular adhesion molecule-1 (ICAM-1), nuclear factor (NF)-κB/p65 and phosphorylated p38 protein in ischemic myocardial tissue of MI rats. In conclusion, gambogic acid exerted anti-inflammatory effects in MI rats by targeting the iNOS, MMPs, ICAM-1, NF-κB and p38 pathways. Gambogic acid may protect against MI-induced inflammation in rats, which may be associated with the activation of the NF-κB/p38 pathway.