Hypertrophy of the right ventricle by pulmonary artery banding in rats: a study of structural, functional, and transcriptomics alterations in the right and left ventricles.

Introduction: Right ventricular remodeling with subsequent functional impairment can occur in some clinical conditions in adults and children. The triggering factors, molecular mechanisms, and, especially, the evolution over time are still not well known. Left ventricular (LV) changes associated with right ventricular (RV) remodeling are also poorly understood. Objectives: The study aimed to evaluate RV morphological, functional, and gene expression parameters in rats submitted to pulmonary artery banding compared to control rats, with the temporal evolution of these parameters, and to analyze the influence of RV remodeling by pulmonary artery banding in rats and their controls over time on LV geometry, histology, gene expression, and functional performance. Methods: Healthy 6-week-old male Wistar-EPM rats weighing 170-200 g were included. One day after the echocardiogram, depending on the animals undergoing the pulmonary artery banding (PAB) procedure or not (control group), they were then randomly divided into subgroups according to the follow-up time: 72 h, or 2, 4, 6, or 8 weeks. In each subgroup, the following were conducted: a new echocardiogram, a hemodynamic study, the collection of material for morphological analysis (hypertrophy and fibrosis), and molecular biology (gene expression). The results were presented as the mean ± standard deviation of the mean. A two-way ANOVA and Tukey post-test compared the variables of the subgroups and evolution follow-up times. The adopted significance level was 5%. Results: There was no significant difference among the subgroups in the percentage of water in both the lungs and the liver (the percentage of water in the lungs ranged from 76% to 78% and that of the liver ranged from 67% to 71%). The weight of the right chambers was significantly higher in PAB animals in all subgroups (RV PAB weighed from 0.34 to 0.48 g, and control subjects, from 0.17 to 0.20 g; right atrium (RA) with PAB from 0.09 to 0.14 g; and control subjects from 0.02 to 0.03 g). In the RV of PAB animals, there was a significant increase in myocyte nuclear volume (97 µm3-183.6 µm3) compared to control subjects (34.2 µm3-57.2 µm3), which was more intense in subgroups with shorter PAB follow-up time, and the fibrosis percentage (5.9%-10.4% vs. 0.96%-1.18%) was higher as the PAB follow-up time was longer. In the echocardiography result, there was a significant increase in myocardial thickness in all PAB groups (0.09-0.11 cm compared to control subjects-0.04-0.05 cm), but there was no variation in RV diastolic diameter. From 2 to 8 weeks of PAB, the S-wave (S') (0.031 cm/s and 0.040 cm/s), and fractional area change (FAC) (51%-56%), RV systolic function parameters were significantly lower than those of the respective control subjects (0.040 cm/s to 0.050 cm/s and 61%-67%). Furthermore, higher expression of genes related to hypertrophy and extracellular matrix in the initial subgroups and apoptosis genes in the longer follow-up PAB subgroups were observed in RV. On the other hand, LV weight was not different between animals with and without PAB. The nuclear volume of the PAB animals was greater than that of the control subjects (74 µm3-136 µm3; 40.8 µm3-46.9 µm3), and the percentage of fibrosis was significantly higher in the 4- and 8-week PAB groups (1.2% and 2.2%) compared to the control subjects (0.4% and 0.7%). Echocardiography showed that the diastolic diameter and LV myocardial thickness were not different between PAB animals and control subjects. Measurements of isovolumetric relaxation time and E-wave deceleration time at the echocardiography were different between PAB animals and control subjects in all subgroups, but there were no changes in diastolic function in the hemodynamic study. There was also increased expression of genes related to various functions, particularly hypertrophy. Conclusion: 1) Rats submitted to pulmonary artery banding presented RV remodeling compatible with hypertrophy. Such alterations were mediated by increased gene expression and functional alterations, which coincide with the onset of fibrosis. 2) Structural changes of the RV, such as weight, myocardial thickness, myocyte nuclear volume, and degree of fibrosis, were modified according to the time of exposure to pulmonary artery banding and related to variations in gene expression, highlighting the change from an alpha to a beta pattern from early to late follow-up times. 3) The study suggests that the left ventricle developed histological alterations accompanied by gene expression modifications simultaneously with the alterations found in the right ventricle.

Improving Enzymatic Saccharification of Peach Palm (Bactris gasipaes) Wastes via Biological Pretreatment with Pleurotus ostreatus.

The white-rot fungus Pleurotus ostreatus was used for biological pretreatment of peach palm (Bactris gasipaes) lignocellulosic wastes. Non-treated and treated B. gasipaes inner sheaths and peel were submitted to hydrolysis using a commercial cellulase preparation from T. reesei. The amounts of total reducing sugars and glucose obtained from the 30 d-pretreated inner sheaths were seven and five times higher, respectively, than those obtained from the inner sheaths without pretreatment. No such improvement was found, however, in the pretreated B. gasipaes peels. Scanning electronic microscopy of the lignocellulosic fibers was performed to verify the structural changes caused by the biological pretreatments. Upon the biological pretreatment, the lignocellulosic structures of the inner sheaths were substantially modified, making them less ordered. The main features of the modifications were the detachment of the fibers, cell wall collapse and, in several cases, the formation of pores in the cell wall surfaces. The peel lignocellulosic fibers showed more ordered fibrils and no modification was observed after pre-treatment. In conclusion, a seven-fold increase in the enzymatic saccharification of the Bactris gasipaes inner sheath was observed after pre-treatment, while no improvement in enzymatic saccharification was observed in the B. gasipaes peel.

Increased Myocardial Retention of Mesenchymal Stem Cells Post-MI by Pre-Conditioning Exercise Training.

Stem cell (SC) therapy is a promising approach to improve post-myocardial infarction (MI) cardiac remodeling, but the proinflammatory microenvironment may lead to SC loss and, therefore, may have a negative impact on therapy. It appears that exercise training (ET) improves myocardial microenvironment for SC transplantation. Therefore, we tested the effect of ET on post-infarction retention of adipose-derived SCs (ADSCs) and its combined effects on the inflammatory microenvironment. Fischer-344 female rats were randomized to one of the following groups: Sham; sedentary coronary occlusion who did not receive ADSCs (sMI); sedentary coronary occlusion who received ADSCs; exercise coronary occlusion who received ADSCs. Rats were trained nine weeks prior to MI, followed by ADSCs transplantation. The MI led to left ventricle (LV) dilation and dysfunction, myocardial hypertrophy and fibrosis, and increased proinflammatory profile compared to Sham rats. Conversely, ADSCs transplanted rats exhibited, better morphological and functional LV parameters; inhibition of myocardial hypertrophy and fibrosis; and attenuation of proinflammatory cytokines (interleukins 1ß and 10, tumor necrosis factor α, and transforming growth factor ß) in the myocardium compared to sMI rats. Interestingly, ET enhanced the effect of ADSCs on interleukin 10 expression. There was a correlation between cytokine expression and myocardial ADSCs retention. The. ET enhanced the beneficial effects of ADSCs in infarcted myocardium, which was associated with higher ADSCs retention. These findings highlight the importance of ET in myocardial retention of ADSCs and attenuation of cardiac remodeling post-infarction. Cytokine analysis suggests improvement in ET-linked myocardial microenvironment based on its anti-inflammatory action.

Photobiomodulation therapy combined with carvedilol attenuates post-infarction heart failure by suppressing excessive inflammation and oxidative stress in rats.

The post-myocardial infarction heart failure (HF) still carries a huge burden since current therapy is unsuccessful to abrogate poor prognosis. Thus, new approaches are needed, and photobiomodulation therapy (PBMt) may be a way. However, it is not known whether PBMt added to a standard HF therapy provides additional improvement in cardiac remodeling in infarcted rats. This study sought to determine the combined carvedilol-drug and PBMt with low-level laser therapy value in HF. Rats with large infarcts were treated for 30 days. The functional fitness was evaluated using a motorized treadmill. Echocardiography and hemodynamic measurements were used for functional evaluations of left ventricular (LV). ELISA, Western blot and biochemical assays were used to evaluate inflammation and oxidative stress in the myocardium. Carvedilol and PBMt had a similar action in normalizing pulmonary congestion and LV end-diastolic pressure, attenuating LV dilation, and improving LV systolic function. Moreover, the application of PBMt to carvedilol-treated rats inhibited myocardial hypertrophy and improved +dP/dt of LV. PBMt alone prevented inflammation with a superior effect than carvedilol. Carvedilol and PBMt normalized 4-hydroxynonenal (a lipoperoxidation marker) levels in the myocardium. However, importantly, the addition of PBMt to carvedilol attenuated oxidized protein content and triggered a high activity of the anti-oxidant catalase enzyme. In conclusion, these data show that the use of PBMt plus carvedilol therapy results in a significant additional improvement in HF in a rat model of myocardial infarction. These beneficial effects were observed to be due, at least in part, to decreased myocardial inflammation and oxidative stress.

Swimming Training Improves Myocardial Mechanics, Prevents Fibrosis, and Alters Expression of Ca2+ Handling Proteins in Older Rats.

Exercise training effects on the contractility of aged myocardium have been investigated for more than 20 years, but the data are still unclear. This study evaluated the hypothesis that a swimming training (ST) may improve myocardial inotropism in older rats. Male Wistar rats aged 4 (young)-and 21 (old)-months-old were divided into young untrained (YNT), old untrained (ONT), and old trained (OTR; 6 weeks of ST) groups. Echocardiography and hemodynamic were employed to assess left ventricular morphology and function. Myocardial mechanics was evaluated on papillary muscles. Histological and immunoblotting were carried out to evaluate fibrosis and proteins that modulate the myocardial function and calcium handling. We found that older rats did not show cardiac dysfunction, but ONT group showed lower physical performance during a swimming test (YNT: 5 ± 2; ONT: -16 ± 0.4; OTR: 51 ± 3; Δ%, sec). Moreover, ONT group showed worse myocardial inotropism, in which it was reversed by ST (Peak developed tension: YNT: 6.2 ± 0.7; ONT: 3.9 ± 0.3; OTR: 6.9 ± 0.9; g/mm2). The ST was associated with preserved collagen content (YNT: 0.38 ± 0.05; ONT: 0.78 ± 0.12; OTR: 0.34 ± 0.09; %). Exercise partially mitigated the effects of aging on intracellular Ca2+-regulating protein (eg, L-Ca2+ channel and phospholamban) and ß-isoform of myosin. Thus, we propose that these molecular alterations together with inhibition of collagen increase contribute to improved myocardial performance in older rats.

Thermotolerance does not reduce the size or remodeling of radiofrequency lesions in the rat myocardium.

PURPOSE: Late lesion extension may be involved in the genesis of delayed radiofrequency (RF) effects. Because RF lesion is thermally mediated, we hypothesized that induction of heat shock response (thermotolerance) would modulate lesion healing. We evaluated the effects of thermotolerance on the dimensions and remodeling of RF lesions in a rat model of heart failure. METHODS: Wistar rats (weight 300 g) subjected to heat stress (n = 22, internal temperature of 42 °C for 10 min) were compared to controls (n = 22, internal temperature of 37 °C for 10 min). After 48 h (peak of HSP70 myocardial concentration), a modified unipolar RF lesion (customized catheter, tip 4.5 mm in diameter; 12 W; 10 s) was created on the left ventricular free wall. Animals were sacrificed 2 h (n = 10 per group) and 4 weeks (n = 12 per group) after ablation for lesion analysis. An echocardiogram was obtained at 4 weeks. RESULTS: There was no difference between groups regarding the size of acute (controls 27 ± 2 vs. treated 27 ± 3 mm(2)) and chronic lesions (controls 17 ± 1 vs. treated 19 ± 1 mm(2)). Histology of lesions did not differ between groups. The echocardiogram revealed dilation of the cavities and moderate systolic dysfunction without difference between groups. Acute lesion dimensions were similar between control and treated animals over time (ablation undertaken 3, 12, 24, 48, and 72 h after hyperthermia) and also using a conventional ablation catheter (50 °C; 15 W; 10 s). CONCLUSION: Thermotolerance does not reduce the size or remodeling of RF lesions in the rat myocardium.

Impact of prophylactic corticosteroids on systemic inflammation after extensive atrial ablation in pigs.

AIMS: Prophylactic corticosteroids have been reported to attenuate the increase in C-reactive protein (CRP) and the incidence of atrial fibrillation (AF) both after heart surgery and AF ablation. We tested the impact of a single prophylactic corticosteroid dose on ultrasensitive CRP 24 h and 14 days after extensive linear atrial ablation (8 mm or 3.5 irrigated tip) guided by electroanatomical mapping (NavX) in pigs with normal hearts. METHODS AND RESULTS: Pigs (n = 19; 35 kg) were divided into three groups: corticoid (n = 7), atrial ablation with administration of 500 mg methylprednisolone intravenous at anaesthetic induction; control (n = 7), atrial ablation only; and sham (n = 5), surgical procedure without ablation. Troponin and CRP were measured before, 24 h and 14 days after the procedure. After sacrifice, lesions were analysed macroscopically and histologically. Linear lesions were created in the right (n = 23) and left (n = 21) atrium of 14 animals, with no difference between groups. In all groups there was elevation of troponin and CRP 24 h after ablation, with a return to baseline values after 14 days. However, CRP levels of the control, corticoid, and sham groups were similar at all three time points analysed (baseline P = 0.52, 24 h P = 0.21, 14 days P = 0.66). Histological analysis did not show any difference between corticoid and control groups. CONCLUSION: In this model, extensive biatrial RF ablation, per se, does not promote systemic inflammation. The use of a prophylactic single corticoid dose before ablation did not prevent systemic inflammation or alter the healing of the lesions.

Impact of corticosteroids on late growth of radiofrequency lesions in infant pigs: histopathological and electroanatomical findings.

AIMS: Corticosteroids attenuate late growth of radiofrequency (RF) lesions in the thigh muscle of infant rats. We sought to assess the impact of these drugs on the late growth of RF lesions in immature swine myocardium and to determine the electroanatomical mapping (EAM) characteristics of these lesions. METHODS AND RESULTS: Radiofrequency (60°C; 60 s) lesions were created in the right atrium (n = 2) and ventricle (n = 2) of 14 piglets (age 65 days; weight 5 kg) and 3 adults. Piglets were divided into: controls (n = 7) and treated (n = 7), receiving hydrocortisone (10 mg/kg iv after RF) and prednisone (1 mg/kg/day) for 29 days. After 8 months, animals were sacrificed for histological analysis. In four piglets, endocardial and epicardial voltage EAM were performed. In infant groups, the dimensions of atrial (11 ± 5 vs. 13 ± 7 mm) and ventricular (12 ± 3 vs. 11 ± 3 mm) lesions were similar. In adults, atrial (6 ± 1 mm) and ventricular (6 ± 1 mm) lesions were smaller. In controls, ventricular lesions depicted dense fibrosis and multiple strands of fibrous tissue extending from the lesion into normal muscle. Treated piglets revealed scars exhibiting less dense fibrosis with predominance of fibroadipose tissue and less collagen proliferation. Large atrial and ventricular low-voltage areas corresponding to the macroscopic lesions were identified in all animals. CONCLUSION: Radiofrequency lesions in infant pigs reveal late growth and invasion of normal muscle by intense collagen proliferation. Corticosteroids do not prevent late enlargement of the lesions but modulate the fibrotic proliferation. The expressive growth of the lesion may generate low-voltage areas detectable by EAM.