RESUMEN
AIM: To investigate whether high-intensity interval training (HIIT) improves physical performance, systolic blood pressure, and markers of oxidative stress and inflammation in skeletal muscle of spontaneously hypertensive rats (SHR). METHODS: Nineteen male SHR rats were randomly assigned to two groups: sedentary (SHRC) and trained (SHR+T). The SHR+T group trained five times a week for eight weeks on a treadmill, while the SHR group remained without any exercise stimulus throughout the experimental period. Maximum physical performance and systolic blood pressure (SBP) were assessed before and after the training period. The following variables were measured in the tibialis anterior (TA) muscle: gene expression of the NADPH oxidase complex (NOX2, NOX4, p22phox, p47phox) and the NF-kB pathway (NF-kB and Ik-B), lipid peroxidation (malonaldehyde; MDA), protein carbonylation, hydrophilic antioxidant capacity (HAC) and pro-inflammatory cytokines (IL-6 and TNF-α). RESULTS: SHR+T rats showed higher physical performance and levels of IL-6, and lower SBP and protein carbonylation (p<0.05), compared with SHRC rats. No significant differences (p>0.05) were observed in the other variables. SIGNIFICANCE: Our results indicate that HIIT is an effective non-pharmacologic strategy to improve physical performance, reduce SBP, and modulate the skeletal muscle oxidative damage and inflammation in hypertensive rats.
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Presión Sanguínea , Entrenamiento de Intervalos de Alta Intensidad , Músculo Esquelético , Estrés Oxidativo , Condicionamiento Físico Animal , Ratas Endogámicas SHR , Animales , Masculino , Músculo Esquelético/metabolismo , Entrenamiento de Intervalos de Alta Intensidad/métodos , Ratas , Biomarcadores/metabolismo , Hipertensión/metabolismo , Hipertensión/fisiopatología , Inflamación/metabolismo , FN-kappa B/metabolismo , NADPH Oxidasas/metabolismo , Interleucina-6/metabolismoRESUMEN
Dysregulated renin-angiotensin system (RAS) signaling contributes to elevated blood pressure (BP), inflammation, and organ damage in systemic arterial hypertension (HTN). We have demonstrated that hypertensive humans and rats exhibit higher expression of classic RAS components and lower expression of counterregulatory RAS components in the lungs compared with normotensive counterparts. Here, we investigated whether BP control could restore the balance between classic [angiotensin I-converting enzyme 2 (ACE)/angiotensin II (Ang II)] and counterregulatory [angiotensin I-converting enzyme 2 (ACE2)/Ang (1-7)] RAS, thereby mitigating lung inflammation. Male spontaneously hypertensive rats (SHRs) were treated with either losartan or amlodipine, both of which effectively reduced BP. These interventions up-regulated lung Ace2 and down-regulated Ace gene expression. Pulmonary membrane ACE2 abundance and activity were higher in losartan- and amlodipine-treated SHRs than in vehicle-treated SHRs, whereas ACE protein and function remained unchanged. Drug-treated SHRs exhibited lower levels of lung Ang II and higher levels of Ang (1-7) than vehicle-treated SHRs. Rebalancing the pulmonary RAS remarkably reduced macrophage number and down-regulated pro-inflammatory genes in SHR lungs, with lower expression of lung pro-inflammatory genes correlating with lower circulating levels of ACE2. Serum analysis in healthy and hypertensive individuals supported these findings, showing higher ACE2 levels in uncontrolled compared with controlled hypertension and normotension. Collectively, these findings suggest that high blood pressure may induce lung inflammation via an ACE/ACE2 imbalance. BP control with either an RAS inhibitor or a calcium channel blocker rebalances RAS in SHR lungs and alleviates inflammation. Furthermore, this study provides a mechanistic link between inflammatory lung diseases (such as COVID-19) and hypertension as a major risk factor.
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Enzima Convertidora de Angiotensina 2 , Antihipertensivos , Presión Sanguínea , Hipertensión , Losartán , Pulmón , Ratas Endogámicas SHR , Sistema Renina-Angiotensina , Animales , Sistema Renina-Angiotensina/efectos de los fármacos , Masculino , Presión Sanguínea/efectos de los fármacos , Pulmón/metabolismo , Pulmón/efectos de los fármacos , Pulmón/fisiopatología , Enzima Convertidora de Angiotensina 2/metabolismo , Enzima Convertidora de Angiotensina 2/genética , Ratas , Hipertensión/fisiopatología , Hipertensión/tratamiento farmacológico , Hipertensión/metabolismo , Losartán/farmacología , Antihipertensivos/farmacología , Antihipertensivos/uso terapéutico , Amlodipino/farmacología , Amlodipino/uso terapéutico , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Humanos , Fragmentos de Péptidos/metabolismo , COVID-19/complicaciones , COVID-19/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Peptidil-Dipeptidasa A/genética , SARS-CoV-2RESUMEN
OBJECTIVE: This study evaluated the effect of Stemregen® nutritional supplement on inflammation and resorption in apical periodontitis using a rat model. METHODOLOGY: Rats were divided in three groups: negative control (n=7), positive control (n=10), and Stemregen® (Stem) (n=10). Apical periodontitis was induced in the positive control and Stem groups, and all rats were sacrificed on the 30th day. Serum phosphorus (P), calcium (Ca), and alkaline phosphatase (ALP) were analyzed. Histopathological assessments measured osteoblastic and osteoclastic activity, inflammation, fibrosis, and abscess density. Immunohistochemical analyses evaluated RANKL, TRAP, and OPG levels. RESULTS: Results showed significantly lower osteoblastic activity in the negative control compared to Stem and positive control groups (p=0.005). Osteoclastic activity was higher in the positive control (p=0.032). Inflammation and abscess formation were reduced in the Stem group compared to the positive control (p<0.001). OPG levels were lower in the negative control compared to the other groups (p=0.005). CONCLUSION: Stemregen® effectively reduced inflammation and bone destruction, suggesting potential benefits for apical periodontitis management, though further research is needed.
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Fosfatasa Alcalina , Modelos Animales de Enfermedad , Osteoprotegerina , Periodontitis Periapical , Ratas Wistar , Animales , Periodontitis Periapical/terapia , Osteoprotegerina/análisis , Masculino , Fosfatasa Alcalina/sangre , Fosfatasa Alcalina/análisis , Calcio/análisis , Calcio/sangre , Fósforo/sangre , Fósforo/análisis , Factores de Tiempo , Reproducibilidad de los Resultados , Resultado del Tratamiento , Ligando RANK/análisis , Inmunohistoquímica , Osteoclastos/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Distribución Aleatoria , Fosfatasa Ácida Tartratorresistente/análisis , Fosfatasa Ácida Tartratorresistente/sangre , RatasRESUMEN
Obesity, a global epidemic, is linked to adverse reproductive outcomes, including infertility and ovulation dysfunction. The cafeteria diet (CAF) serves as an animal model mirroring Western diet habit. Coenzyme Q10 (CoQ10), known for enhancing reproductive outcomes in various pathologies, is not fully understood for its effects on obesity treatment. Here, obesity was modeled using CAF-fed rats to assess CoQ10's impact on metabolic and ovarian disruptions caused by obesity. Wistar rats were divided into control (standard diet) and obese (CAF diet) groups. After 75 days, half of each group received oral CoQ10 (5 mg/kg) for 13 days, while the rest received a vehicle. Animals were euthanized during the estrus phase, and blood and ovaries were collected for analysis. CAF caused increased body weight gain (p < 0.01) associated with hyperglycemia, hypertriglyceridemia, and hypercholesterolemia (p < 0.05). Moreover, it caused a reduction in the number of AMH + follicles (p < 0.001), increasing follicular atresia (p < 0.05) and serum estradiol levels (p < 0.05). Obesity also altered the estrous cycle and reduced the ovulation rate (p < 0.05). CoQ10 administration showed beneficial effects on all ovarian disruptions but had no effect on the metabolic alterations induced by obesity. In summary, CoQ10 could be an additional treatment for obesity-related infertility in patients with normal metabolic profiles. While CoQ10 does not affect metabolic parameters influenced by obesity, crucial for reproductive issues and offspring health, it is recommended as part of a treatment plan that includes a balanced diet and increased physical activity for obese individuals with metabolic alterations seeking pregnancy.
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Suplementos Dietéticos , Obesidad , Ratas Wistar , Reproducción , Ubiquinona , Animales , Ubiquinona/análogos & derivados , Ubiquinona/farmacología , Ubiquinona/uso terapéutico , Femenino , Obesidad/tratamiento farmacológico , Obesidad/metabolismo , Ratas , Reproducción/efectos de los fármacos , Ovario/efectos de los fármacos , Ovario/metabolismoRESUMEN
OBJECTIVES: To carry out physicomechanical characterization of the HA/DCPA/TiO2 and to evaluate the tissue repair in rat calvaria. METHODS: Two bone defects were made in the calvaria of 36 Wistar rats, divided into groups: HA/DCPA, HA/DCPA/TiO2 and sham (blood clot). The animals were euthanized at 30, 60 and 90 days and calvaria slides were processed with hematoxylin/eosin. The newly formed bone, connective tissue, biomaterial remnant, and total tissue repair percentages were calculated in relation to the total defect area. The HA/DCPA/TiO2 was characterized structurally by scanning electron microscopy (SEM), and chemically by energy-dispersive X-ray spectroscopy (EDS) and X-ray diffraction (XRD). It was submitted to apparent density (AD), apparent porosity (AP), water absorption (WA) and compressive strength (CS) physical tests. The ANOVA test was applied, followed by Turkey's test and Student's t-test (p ≤ 0,05). RESULTS: The SEM showed biomaterials inside the bone defects and newly formed bone. EDS identified oxygen, calcium, phosphorus, and titanium in the sample. The HA/DCPA/TiO2 and HA/DCPA groups presented a total tissue repair area that was larger than the sham group (p < 0.001). CONCLUSIONS: The physical-mechanical assays showed that HA/DCPA/TiO2 has AD and CS properties within the limits of trabecular bone and with values higher than HA/DCPA.HA/DCPA/TiO2 presented higher densification and compressive strength rates than HA/DCPA. CLINICAL RELEVANCE: Potential as a scaffold for bone.
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Microscopía Electrónica de Rastreo , Ratas Wistar , Cráneo , Titanio , Animales , Titanio/química , Ratas , Cráneo/cirugía , Difracción de Rayos X , Cáscara de Huevo/química , Espectrometría por Rayos X , Regeneración Ósea/efectos de los fármacos , Ensayo de Materiales , Materiales Biocompatibles/farmacología , Materiales Biocompatibles/química , Masculino , Fuerza Compresiva , Bioingeniería , Fosfatos de Calcio/química , Fosfatos de Calcio/farmacología , Propiedades de Superficie , Porosidad , Durapatita/farmacología , Durapatita/químicaRESUMEN
Sirtuins (SIRTs) are key regulators of cellular metabolism, involved in a wide range of physiological and pathological processes. However, there is scarce knowledge about the effect of sugar consumption and physical activity on SIRTs in kidney disorders. Here, we evaluated the impact of prolonged consumption of an isocaloric high-sugar diet (HSD) and physical training on the modulation of renal Sirts and the link between these alterations and possible obesity-associated kidney damage. Newly weaned male Wistar rats were fed a standard chow diet (STD) or HSD ad libitum and then subjected or not to regular workload swimming training for 18 weeks. Morphometric and biochemical parameters were analyzed, and the kidneys were removed for lipid quantification, histological analysis, and for Sirts1-7 expression. HSD led to the development of obesity, increased serum triglyceride levels, and glucose intolerance, regardless of higher caloric consumption. However, training was able to partially inhibit the HSD-induced obesogenic effect. No changes were identified in kidney mass, lipid content, histology, and creatinine clearance among the groups; these results were associated with a decrease in the renal expression of Sirt2-3 and Sirt7; however, training was able to reverse this modulation. The interaction between HSD and training led to an increase in Sirt4-7. However, Sirt1 remained constant among experimental groups. In conclusion, our results indicated that the transcriptional modulation of Sirts precedes HSD-induced damage and loss of kidney function, as well as a possible protective adaptive response of physical exercise on long-term Sirts expression.
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Condicionamiento Físico Animal , Ratas Wistar , Sirtuinas , Natación , Animales , Masculino , Natación/fisiología , Condicionamiento Físico Animal/fisiología , Sirtuinas/metabolismo , Ratas , Obesidad/metabolismo , Riñón/metabolismoRESUMEN
Alpinia zerumbet, a plant native to East Asia, is widely found on the Brazilian coast, where it is used in folk medicine as an antihypertensive, diuretic, and anxiolytic. This study investigated the effects of the hydroalcoholic extract obtained from Alpinia zerumbet leaves (AZE) on cardiovascular changes and oxidative status in spontaneously hypertensive rats (SHR). SHR and Wistar-Kyoto male rats, 90 days old, treated or not with AZE (50 mg/kg/day in drinking water) for six weeks, were used in this study. Blood pressure (BP) was assessed weekly by tail plethysmography. At the end of treatment, the animals were anesthetized with thiopental (70 mg/kg, ip), blood was collected through abdominal aorta puncture, the thoracic aorta and left ventricle were isolated for morphometric analysis and immunostaining of NOX-4, SOD-2, 8-isoprostane, and angiotensin II AT1 receptors (AT1R), and the mesenteric arterial bed (MAB) was isolated for the assessment of vascular function. Oxidative damage in lipids and proteins and the enzymatic antioxidant activity were evaluated in plasma samples by spectrophotometry. AZE normalized BP in SHR. Although the treatment did not improve the MAB vascular dysfunction, it reversed the cardiovascular remodeling in the aorta and left ventricle. In addition, AZE improved antioxidant activity in plasma and SOD-2 immunostaining in the thoracic aorta and left ventricle, decreased protein carbonylation in plasma, and reduced 8-isoprostane, NOX-4, and AT1R immunostaining in the cardiovascular system. The results suggested that AZE reversed hypertension and cardiovascular remodeling in SHR, which was associated with lower oxidative stress and AT1R.
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Alpinia , Antihipertensivos , Hipertensión , Extractos Vegetales , Hojas de la Planta , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Animales , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Hipertensión/tratamiento farmacológico , Hipertensión/fisiopatología , Masculino , Alpinia/química , Ratas , Hojas de la Planta/química , Antihipertensivos/farmacología , Presión Sanguínea/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Ventrículos Cardíacos/efectos de los fármacos , Ventrículos Cardíacos/fisiopatología , Remodelación Ventricular/efectos de los fármacosRESUMEN
Necrotizing enterocolitis (NEC) is a severe intestinal disease of multifactorial origin that primarily affects premature infants. Approximately 27% of NEC babies develop short gut (SG) secondary to extensive intestinal resection, and 10% will have chronic dependence on total parenteral nutrition. We evaluated the Botox treatment in SG model rats. Twenty-day-old weanling male rats (weight range 38-70 g, n=72) were divided into four groups (n=18 each): 1) Control (fed a regular liquid diet); 2) Botox (Control submitted to laparotomy and intestinal injection of Botox®); 3) SG (short gut); and 4) SG and Botox (SG+Botox®). After seven post-operative days, samples were collected for biometrics [body weight (BW), intestine weight (IW) and IW/BW ratio (IBR), and intestine length (IL) and height (IH)], histometric analysis [villous height (VH), crypt depth (CD), muscular thickness (MT), and PCNA index)], and intestinal transit time (ITT). BW, IW, and IL decreased in SG (P<0.05). IH, VH, and PCNA index increased in Botox groups [Control = SG < Botox and SG+Botox (P<0.05)], CD increased in Botox, SG, and SG+Botox (P<0.005), and MT was higher in SG and SG+Botox. Botox groups had lower ITT (P<0.05). Botox provided dilatation and histological changes in SG. These findings suggested that Botox improved adaptation and might be applied in SG with promising results.
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Adaptación Fisiológica , Síndrome del Intestino Corto , Animales , Masculino , Síndrome del Intestino Corto/tratamiento farmacológico , Adaptación Fisiológica/efectos de los fármacos , Ratas , Modelos Animales de Enfermedad , Ratas Wistar , Toxinas Botulínicas Tipo A/farmacología , Toxinas Botulínicas Tipo A/administración & dosificación , Enterocolitis Necrotizante/tratamiento farmacológico , Animales Recién Nacidos , Destete , Intestinos/efectos de los fármacosRESUMEN
Cancer is the second leading cause of death worldwide. Cancer cachexia is a multifactorial catabolic syndrome responsible for almost one third of cancer-related deaths. Drug repurposing has been used in oncological research and drugs like clenbuterol and metformin seem to be reasonable candidates in the context of cancer cachexia, because the former is a ß2-agonist that stimulates muscle gain and the latter has anti-inflammatory properties. The aim of this study was to assess the effects of a short-term treatment with metformin and clenbuterol, isolated or combined, on tumor growth and cancer cachexia parameters in Walker 256 tumor-bearing rats, a model of cancer cachexia. To this end, Wistar rats were separated into 8 groups and 4 of them were injected with Walker 256 tumor cells (W groups). Control (C) and W groups received the following treatments: metformin (M), clenbuterol (Cb), or metformin combined with clenbuterol (MCb). Body and tumor weight, metabolic parameters, and oxidative damage in the tumor were assessed. Compared to the C group, the W group showed body weight loss, hypoglycemia, hyperlactatemia, and hypertriacylglycerolemia. None of the treatments could reverse body weight loss, although they reversed the alterations of the assessed plasma metabolic parameters. Surprisingly, only clenbuterol alone reduced tumor weight. Hydrogen peroxide production and lipid peroxidation in tumor tissue was increased in this group. In conclusion, metformin and clenbuterol ameliorated metabolic cachexia parameters in Walker tumor-bearing rats, but only clenbuterol reduced the tumor weight, probably, through a lipid peroxidation-dependent cell death.
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Caquexia , Carcinoma 256 de Walker , Clenbuterol , Peroxidación de Lípido , Metformina , Ratas Wistar , Animales , Caquexia/tratamiento farmacológico , Caquexia/etiología , Metformina/farmacología , Metformina/uso terapéutico , Clenbuterol/farmacología , Clenbuterol/uso terapéutico , Carcinoma 256 de Walker/tratamiento farmacológico , Carcinoma 256 de Walker/complicaciones , Masculino , Peroxidación de Lípido/efectos de los fármacos , Ratas , Hipoglucemiantes/farmacología , Hipoglucemiantes/uso terapéutico , Modelos Animales de Enfermedad , Agonistas Adrenérgicos beta/farmacologíaRESUMEN
Melatonin is a pineal hormone synthesized exclusively at night, in several organisms. Its action on sperm is of particular interest, since they transfer genetic and epigenetic information to the offspring, including microRNAs, configuring a mechanism of paternal epigenetic inheritance. MicroRNAs are known to participate in a wide variety of mechanisms in basically all cells and tissues, including the brain and the sperm cells, which are known, respectively, to present 70% of all identified microRNAs and to transfer these molecules to the embryo. MicroRNAs from sperm have been associated with modulation of embryonic development and inheritance of psychiatric symptoms, including autism. Given that microRNAs and melatonin are ubiquitous molecules with important roles in the organism, the aim of this study was to investigate the expression of specific microRNAs in sperm, brain and cerebellum of pinealectomized rats. For this study, Wistar rats had their pineal gland removed at 60 post-partum. Part of these rats received exogenous melatonin until the day of the euthanasia. The control group did not receive any treatment or manipulation. The sperm, hypothalamus, prefrontal cortex and cerebellum were collected for analysis of microRNA expression by RT-qPCR. The results suggest that melatonin absence caused by pinealectomy increases the expression of the target microRNAs in the sperm. Although the data suggest an alteration (increase or decrease depending on the region and microRNA) of expression levels of some microRNAs in the brain and cerebellum of pinealectomized rats, the differences were not statistically significant. This seems to be a consequence of the intragroup variation. Melatonin administration restored the levels of the target microRNAs in the sperm. Additional studies are needed to understand the impact of the alterations of microRNA expression to the pinealectomized rats as well as to their descendants.
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Cerebelo , Hipotálamo , Melatonina , MicroARNs , Corteza Prefrontal , Ratas Wistar , Espermatozoides , Animales , Melatonina/farmacología , MicroARNs/genética , MicroARNs/metabolismo , Masculino , Ratas , Corteza Prefrontal/metabolismo , Corteza Prefrontal/efectos de los fármacos , Cerebelo/metabolismo , Cerebelo/efectos de los fármacos , Espermatozoides/metabolismo , Espermatozoides/efectos de los fármacos , Hipotálamo/metabolismo , Hipotálamo/efectos de los fármacos , Glándula Pineal/metabolismo , Glándula Pineal/cirugía , Pinealectomía , Regulación de la Expresión Génica/efectos de los fármacosRESUMEN
Partial stenosis of the renal artery causes renovascular hypertension (RVH) and is accompanied by chronic renal ischemia, resulting in irreversible kidney damage. Revascularization constitutes the most efficient therapy for normalizing blood pressure (BP) and has significant benefits for renal function; however, the tissue damage caused by chronic hypoxia is not fully reversed. Mesenchymal stem cells (MSCs) have produced discrete results in minimizing RVH and renal tissue and functional improvements since the obstruction persists. This study aimed to evaluate the effects of administration of MSCs in combination with renal artery revascularization in rats subjected to RVH. The following groups were evaluated: control (SHAM), hypertensive (2K-1C), hypertensive treated with MSCs (MSC), hypertensive subjected to revascularization (REV), and hypertensive subjected to revascularization and treatment with MSCs (REV + MSC). The animals were followed up for 10 weeks. The animals in the MSC group received cell infusions at the 3rd, 5th, 7th and 9th weeks. In the REV and REV + MSC groups, the clip was removed by the 6th week (revascularization), and in the REV + MSC group, MSCs infusion was performed at the 6th and 8th weeks. Tail systolic blood pressure (SBP) was measured weekly, and histological parameters and renal function were evaluated at the end of the protocol. The clipped animals developed RVH, deterioration of total renal function (50% decrease in creatinine clearance), and significant proteinuria (15x increase). Treatment with MSCs had no detectable beneficial effects on kidney function or SBP. REV resulted in normalization of BP and a significant but partial reduction in proteinuria (80% vs. 2K-1C), but areas with renal fibrosis persisted. The combination of the two treatments was effective at normalizing all renal parameters as well as reversing proteinuria, reducing the number of ischemic glomeruli and atrophic tubules, indicating an improvement of the renal parenchyma. The results suggest that therapy with MSCs associated with revascularization can potentially help in the full recovery of renal function in the long term in patients with RVH.
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Modelos Animales de Enfermedad , Hipertensión Renovascular , Riñón , Trasplante de Células Madre Mesenquimatosas , Arteria Renal , Animales , Hipertensión Renovascular/terapia , Hipertensión Renovascular/fisiopatología , Hipertensión Renovascular/cirugía , Ratas , Trasplante de Células Madre Mesenquimatosas/métodos , Masculino , Riñón/patología , Riñón/fisiopatología , Arteria Renal/cirugía , Presión Sanguínea , Células Madre Mesenquimatosas/citología , Ratas WistarRESUMEN
BACKGROUND: Cavernous nerve injury-induced erectile dysfunction (CNI-ED) is a common complication following radical prostatectomy and severely affects patients' quality of life. The mitochondrial impairment in corpus cavernosum smooth muscle cells (CCSMCs) may be an important pathological mechanism of CNI-ED. Previous studies have shown that transplantation of human adipose derived stem cells (ADSC) can alleviate CNI-ED in a rat model. However, little is known about the effect of human umbilical cord mesenchymal stem cells (hUC-MSC) on CNI-ED. It remains unclear whether hUC-MSC can ameliorate mitochondrial damage in CCSMCs. In this study, we aimed to investigate the impacts of hUC-MSC on the mitochondrial mass and function of CCSMCs, as well as elucidate its underlying molecular mechanism. METHODS: The CNI-ED rat model was established by bilaterally crushing cavernous nerves. Subsequently, hUC-MSC were transplanted into the cavernosum and ADSC were injected as a positive control group. Erectile function evaluation and histological detection were performed 4 weeks after cell transplantation. In vitro, CCSMCs underwent hypoxia and were then co-cultured with ADSC or hUC-MSC using a transwell system. The mitochondrial mass and function, as well as signaling pathways, were investigated. To explore the role of the SIRT1/PGC-1α/TFAM pathway in regulating mitochondrial biogenesis of CCSMCs, we knocked down SIRT1 by siRNA. RESULTS: The administration of hUC-MSC significantly improved erectile function of CNI-ED rats and reduced the ratio of collagen to smooth muscle. Specifically, hUC-MSC treatment restored mitochondrial mass and function in CCSMCs injured by CNI or hypoxia, and inhibited the apoptosis of CCSMCs. Mechanistically, the application of hUC-MSC activated SIRT1/PGC-1α/TFAM pathway both in rat penile tissues and CCSMCs. In addition, knockdown of SIRT1 in CCSMCs abolished the protective effects of hUC-MSC on mitochondrial mass and function, while leading to an increase in cellular apoptosis. CONCLUSIONS: hUC-MSC contribute to the recovery of erectile function in CNI-ED rats by restoring mitochondrial mass and function of CCSMCs through the SIRT1/PGC-1α/TFAM pathway. Our present study offers new insights into the role and molecular mechanisms of hUC-MSC in regulating mitochondrial homeostasis, thereby facilitating the restoration of the erectile function in CNI-ED.
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Modelos Animales de Enfermedad , Disfunción Eréctil , Mitocondrias , Pene , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma , Ratas Sprague-Dawley , Transducción de Señal , Sirtuina 1 , Masculino , Animales , Sirtuina 1/metabolismo , Disfunción Eréctil/etiología , Disfunción Eréctil/terapia , Ratas , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Pene/inervación , Miocitos del Músculo Liso , Humanos , Factores de Transcripción/metabolismo , Células Madre Mesenquimatosas , Erección Peniana/fisiología , Traumatismos de los Nervios Periféricos/terapia , Trasplante de Células Madre Mesenquimatosas/métodos , Proteínas Mitocondriales/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Argemone mexicana L. (Papaveraceae), a weed that thrives in the tropical and subtropical areas of South and Central America, Mexico, Caribbean Islands and India. In India, it has been used traditionally to treat vesicular calculus, inflammatory conditions, and hepatobiliary disorders. AIM OF THE STUDY: The present study was aimed to investigate the hepatoprotective efficacy of A. Mexicana roots in paracetamol (PCM)-induced toxicity rat. MATERIALS AND METHODS: The methanol extract of A. mexicana (MEAM) root was analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) analysis to identify its compounds. Molecular docking analysis of the compounds was conducted against TGF-ß and PPAR-α. The hepatoprotective activity of MEAM (200, 400 mg/kg) was evaluated in PCM (3000 mg/kg) intoxicated rats by measuring serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), total bilirubin (TB), total protein (TP), albumin (ALB), globulin (GLB), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). Silymarin (100 mg/kg) was used as reference drug. Oxidative stress biomarkers such as superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and lipid peroxidation (LPO) were investigated using liver homogenate. Additionally, the levels of pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and anti-inflammatory cytokines interleukin-4 (IL-4) and interleukin-10 (IL-10) were studied. The results of the study were supported by histopathological examination. RESULTS: GC-MS analysis revealed 163 compounds, from which eleven compounds were selected based on their docking scores against TGF-ß and PPAR-α. MEAM (400 mg/kg) demonstrated a remarkable reduction in ALT, AST, ALP, GGT, and LDH in contrast to the PCM intoxicated group. A remarkable decline in TB and GLB, along with an increase in TP and ALB, was observed in the MEAM (400 mg/kg) group compared to the untreated PCM group. Rats receiving MEAM (400 mg/kg) exhibited a noticeable decrease in TC, TG, and LDL-C, along with an increase in HDL-C levels compared to PCM-induced untreated rats. The higher dose of MEAM also resulted in a significant decrease in TNF-α, IL-1ß, and IL-6, and an increase in IL-4 and IL-10. Similarly, a notable elevation in SOD, CAT, and GSH, along with a decrease in MDA content, was observed in the group receiving MEAM (400 mg/kg). The histopathological result showed reduction of sinusoidal space and vesicular nuclei, with improvement of hepatocytes at the dose of MEAM (400 mg/kg). In molecular docking study, Eupatilin exhibited the highest docking scores of -10.4 kcal/mol and -9.1 kcal/mol against TGF-ß and PPAR-α, respectively. CONCLUSIONS: MEAM root at dose of 400 mg/kg exhibited hepatoprotective effect against PCM-induced toxicity rat. Eupatilin might be considered as a potential candidate for the hepatoprotective effect of A. mexicana root.
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Acetaminofén , Enfermedad Hepática Inducida por Sustancias y Drogas , Simulación del Acoplamiento Molecular , Extractos Vegetales , Raíces de Plantas , Ratas Wistar , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Acetaminofén/toxicidad , Extractos Vegetales/farmacología , Masculino , Raíces de Plantas/química , Ratas , Hígado/efectos de los fármacos , Hígado/patología , Hígado/metabolismo , Estrés Oxidativo/efectos de los fármacos , Modelos Animales de Enfermedad , Antioxidantes/farmacología , Sustancias Protectoras/farmacologíaRESUMEN
BACKGROUND: Intestinal transplantation (ITx) represents the only curative option for patients with irreversible intestinal failure. Nevertheless, its rejection rate surpasses that of other solid organ transplants due to the heightened immunological load of the gut. Regulatory T-cells (Tregs) are key players in the induction and maintenance of peripheral tolerance, suggesting their potential involvement in modulating host vs. graft responses after ITx. Thus, we investigated the association of Tregs with allograft outcomes in pediatric patients and in an experimental model of small bowel transplantation. METHODS: Treg frequency in human samples was analyzed by Flow cytometry (CD4+CD25highCD127-, blood samples) and immunohistochemistry (FoxP3, graft samples). Experimental allogenic-heterotopic small bowel transplantation was performed in rats and animals divided into 3 groups: non-immunosuppressant treatment, rapamycin (2 mg/kg), and tacrolimus (0.6 mg/kg) treatment. Acute cellular rejection (ACR) was diagnosed based on clinical and histological findings, graft gene expression of pro- and anti-inflammatory mediators assessed by RT-qPCR, serum IL-6 and IL-10 levels by Luminex, and Treg frequency analyzed by flow cytometry (CD4+CD25highFoxP3+). RESULTS: Blood samples from patients undergoing ACR exhibited a significant reduction in the Treg number compared to those with normo-functional grafts. Similarly, a diminished number of FoxP3+ cells was observed in mucosa samples with ACR. In the experimental model, rapamycin-treated animals displayed clinical and histological findings resembling those not receiving immunosuppression treatment. Notably, ACR correlated with a high CD8/CD4 ratio, loss of T-cell chimerism, mRNA upregulation of pro-inflammatory genes and diminished graft Treg frequency. In contrast, tacrolimus treatment prevented ACR and facilitate blood and graft Treg expansion. Remarkably, recipients who achieved Treg expansion within the graft remained free of ACR even after discontinuation of the immunosuppressant treatment and this phenomenon was associated with increased levels of serum IL-10. CONCLUSION: Our clinical and experimental findings underscore the association between Treg frequency and graft rejection after ITx, advocating for strategies that promote their expansion within the gut mucosa to enhance long-term outcomes.
Asunto(s)
Rechazo de Injerto , Intestino Delgado , Linfocitos T Reguladores , Linfocitos T Reguladores/inmunología , Rechazo de Injerto/inmunología , Animales , Intestino Delgado/trasplante , Intestino Delgado/inmunología , Humanos , Masculino , Ratas , Femenino , Niño , Adolescente , Preescolar , Inmunosupresores/uso terapéutico , Tacrolimus/uso terapéutico , Tacrolimus/farmacología , Sirolimus/farmacología , Sirolimus/uso terapéuticoRESUMEN
Childhood obesity increases the risk of developing metabolic diseases in adulthood, since environmental stimuli during critical windows of development can impact on adult metabolic health. Studies demonstrating the effect of prepubertal diet on adult metabolic disease risk are still limited. We hypothesized that a prepubertal control diet (CD) protects the adult metabolic phenotype from diet-induced obesity (DIO), while a high-fat diet (HFD) would predispose to adult metabolic alterations. Sprague-Dawley male rats were fed either a CD or a HFD during the prepubertal period (day 30-40 of age) and subsequently a chronic HFD or CD, respectively, until adulthood (day 220 of age). As controls, rats aged 30 days were exclusively fed a CD or a HFD until adulthood. Body weight and composition, metabolic rate, biochemical and hormonal plasma measurements, hepatic gene expression and methylation and hydroxymethylation levels were analyzed at ages 30, 40 and 220 days. The prepubertal CD prevented fat mass accumulation, lean mass loss and metabolic inflexibility, showed lower insulin, leptin and cholesterol concentrations in adulthood despite the chronic HFD. Notably, the prepubertal CD led to higher hepatic Lxrα expression, lower hepatic global DNA methylation and higher hydroxymethylation in adulthood despite a chronic HFD. Conversely, a prepubertal HFD decreased adult metabolic flexibility, increased serum cholesterol, and decreased Lxrα expression and global DNA hydroxymethylation, while also increasing DNA methylation levels despite a chronic CD. In summary, a prepubertal CD protected the adult metabolic phenotype from high cholesterol concentrations associated with increased hepatic Lxrα expression and lower hepatic global DNA methylation in adulthood, despite exposure to a chronic HFD. Conversely, a prepubertal HFD altered the adult metabolic phenotype.
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Colesterol , Metilación de ADN , Dieta Alta en Grasa , Receptores X del Hígado , Hígado , Ratas Sprague-Dawley , Animales , Masculino , Metilación de ADN/efectos de los fármacos , Receptores X del Hígado/metabolismo , Receptores X del Hígado/genética , Hígado/metabolismo , Ratas , Colesterol/sangre , Colesterol/metabolismo , Dieta Alta en Grasa/efectos adversos , Receptores Nucleares Huérfanos/metabolismo , Receptores Nucleares Huérfanos/genética , Obesidad/metabolismo , Obesidad/etiología , Peso Corporal/efectos de los fármacosRESUMEN
Epilepsy is a chronic neurological disorder that affects nearly 50 million people worldwide. Experimental evidence suggests that epileptic neurons are linked to the endocannabinoid system and that inhibition of the FAAH enzyme could have neuroprotective effects by increasing the levels of endogenous endocannabinoid anandamide. In this context, the use of macamides as therapeutic agents in neurological diseases has increased in recent years. With a similar structure to anandamide, several theories point to the FAAH-macamide interaction as a possible cause of FAAH enzymatic inhibition. In this work, we used in silico and in vivo techniques to analyze the potential therapeutic effect of three synthetic macamides in the treatment of epilepsy: N-3-methoxybenzyl-oleamide (3-MBO), N-3-methoxybenzyl-linoleamide (3-MBL), and N-3-methoxybenzyl-linolenamide (3-MBN). In the first stage, an in silico analysis was conducted to explore the energetic affinity of these macamides with rFAAH and their potential inhibitory effect. MD simulations, molecular docking, and MM/PBSA calculations were used for these purposes. Based on our results, we selected the two best macamides and performed an in vivo study to analyze their therapeutic effect in male Sprague Dawley rat models. Rats were subjected to an in vivo induction of epileptic status by the intraperitoneal injection of pilocarpine and analyzed according to the Racine scale. In silico results showed an energetic affinity of three macamides and a possible "plugging" effect of the membrane access channel to the active site as a potential cause of FAAH inhibition. On the other hand, the in vivo results showed an anticonvulsant effect of both macamides, with 3-MBL being the most active, resulting in a higher survival probability in the rats. This work represents one of the first studies on the use of macamides for the treatment of epilepsy.
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Anticonvulsivantes , Epilepsia , Simulación del Acoplamiento Molecular , Animales , Anticonvulsivantes/farmacología , Anticonvulsivantes/química , Ratas , Epilepsia/tratamiento farmacológico , Epilepsia/inducido químicamente , Masculino , Simulación de Dinámica Molecular , Amidohidrolasas/antagonistas & inhibidores , Amidohidrolasas/metabolismo , Simulación por Computador , Alcamidas Poliinsaturadas/farmacología , Alcamidas Poliinsaturadas/química , Modelos Animales de Enfermedad , Ratas Sprague-DawleyRESUMEN
Diabetes mellitus is a metabolic syndrome that has grown globally to become a significant public health challenge. Hypothesizing that the plasma membrane protein, transient receptor potential ankyrin-1, is a pivotal target in insulin resistance, we investigated the mechanism of action of cinnamaldehyde (CIN), an electrophilic TRPA1 agonist, in skeletal muscle, a primary insulin target. Specifically, we evaluated the effect of CIN on insulin resistance, hepatic glycogen accumulation and muscle and adipose tissue glucose uptake. Furthermore, the in vitro role of CIN in glucose uptake and intracellular signaling was determined in insulin-resistant rats whose calcium influx was analyzed. Moreover, the serum lipid profile was assessed following short-term CIN treatment in rats, and lipid tolerance was analyzed. The effects of CIN on insulin resistance were mediated by TRPA1, with downstream signaling involving the activation of PI3-K, MAPK, PKC, as well as extracellular calcium and calcium release from intracellular stores. Additionally, cytoskeleton integrity was required for the complete action of CIN on glucose uptake in muscle. CIN also ameliorated the serum lipid profile and improved triglyceride tolerance following acute vivo exposure.
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Acroleína , Glucosa , Homeostasis , Resistencia a la Insulina , Metabolismo de los Lípidos , Músculo Esquelético , Canal Catiónico TRPA1 , Animales , Acroleína/análogos & derivados , Acroleína/farmacología , Masculino , Homeostasis/efectos de los fármacos , Ratas , Músculo Esquelético/metabolismo , Músculo Esquelético/efectos de los fármacos , Glucosa/metabolismo , Metabolismo de los Lípidos/efectos de los fármacos , Canal Catiónico TRPA1/metabolismo , Calcio/metabolismo , Transducción de Señal/efectos de los fármacos , Ratas Wistar , Lípidos/sangre , Hígado/metabolismo , Hígado/efectos de los fármacosRESUMEN
Doxorubicin-induced cardiomyopathy (DOX-IC) is a significant and common complication in patients undergoing chemotherapy, leading to cardiac remodeling and reduced heart function. We hypothesized that the intrapericardial injection of hydrogels derived from the cardiac decellularized extracellular matrix (dECM) loaded with adipose tissue-derived stromal cells (ASC) and their secretome dampens or reverses the progression of DOX-IC. DOX-IC was induced in Wistar male rats through ten weekly intra-peritoneal injections of doxorubicin (cumulative dose: 18 mg/kg). We performed intrapericardial treatment in week five with dECM hydrogel loaded with ASC and their conditioned medium (CMed). The volume of intrapericardial injection was 2 ml/kg, the ASC density was 20 million/mL, while the hydrogel contained 100-fold concentrated CMed. Interstitial myocardial fibrosis was assessed by PicroSirius Red staining and hemodynamics parameters in pressure-volume loops. Compared to saline controls, interstitial myocardial fibrosis was reduced in ASC/CMed-loaded hydrogels treated animals (p = 0.0139). Ejection fraction and cardiac work efficiency improved in the ASC/CMed-treated rats compared to saline treatment (p = 0.0151 and p = 0.0655, respectively). The intrapericardial injection of dECM hydrogels loaded with ASC and their secretome warrants a novel therapeutic modality to improve ventricular hemodynamics and reduce cardiac remodeling in DOX-IC.
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Cardiomiopatía Dilatada , Doxorrubicina , Hidrogeles , Ratas Wistar , Animales , Hidrogeles/química , Masculino , Ratas , Doxorrubicina/administración & dosificación , Cardiomiopatía Dilatada/tratamiento farmacológico , Cardiomiopatía Dilatada/patología , Cardiomiopatía Dilatada/metabolismo , Secretoma , Tejido Adiposo/metabolismo , Matriz Extracelular/metabolismo , Fibrosis , Células del Estroma/metabolismo , Miocardio/metabolismo , Miocardio/patología , Modelos Animales de Enfermedad , Pericardio/efectos de los fármacosRESUMEN
This study evaluated the osteogenic potential of the bioactive glasses SinGlass (45S5) and SinGlass High (F18) in regenerating critical bone defects in rat calvaria. Both biomaterials promoted new bone formation around the particles, with the SinGlass High (F18) group exhibiting a higher rate of bone maturation. Histomorphological and birefringence analyses revealed better organization of the newly formed bone in the biomaterial-treated groups, and immunohistochemistry indicated the expression of osteogenic markers such as osteocalcin, immunostaining for bone morphogenetic protein 2 (BMP 2), and immunostaining for bone morphogenetic protein 4 (BMP 4). Microtomography computadorized (Micro-CT) revealed centripetal bone formation in both groups, with greater integration of the particles into the surrounding bone tissue. The superior performance of SinGlass High (F18) was attributed to its higher potassium and magnesium content, which enhance osteoconductivity. After 42 days, the SinGlass High (F18) group showed the highest percentage of new bone formation, in line with previous studies. Although our results are promising, the limited follow-up period and use of a single animal model highlight the need for further research to validate clinical applicability. SinGlass High (F18) appears to be a viable alternative to autografts in bone repair, with potential to improve tissue integration and accelerate recovery.
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Regeneración Ósea , Vidrio , Osteogénesis , Animales , Vidrio/química , Ratas , Regeneración Ósea/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Masculino , Cerámica/química , Cerámica/farmacología , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Microtomografía por Rayos X , Cráneo/efectos de los fármacos , Cráneo/metabolismo , Cráneo/patología , Ratas WistarRESUMEN
Hypothyroidism causes ovarian dysfunction and infertility in women and animals and impairs the hypothalamic expression of kisspeptin (Kp). However, kisspeptin is also expressed in the genital system, and the lack of the Kp receptor (Kiss1r) in the uterus is linked to reduced implantation rates. This study investigated the impact of hypothyroidism on the uterine expression of Kp and Kiss1r in female rats throughout the estrous cycle and the associated changes in uterine activity modulators. Hypothyroidism was induced through daily administration of propylthiouracil (PTU) over a period of 14 days. Plasma levels of LH, E2, and P4, cyclicity, body and uterine weight, uterine histomorphometry, and the gene and/or protein expression of Kiss1, Kiss1r, estrogen receptor α (ERα), progesterone receptor (PR), and thyroid hormone receptor α (TRα) were assessed. Additionally, proliferative activity (CDC-47) and the gene expression of uterine receptivity mediators (SMO, WNT4, BMP2, HAND2, MUC1, and LIF) were evaluated. Hypothyroidism prolonged the diestrus and increased progesterone levels during this phase, while decreasing luteinizing hormone and estradiol on proestrus. In the uterus, hypothyroidism reduced Kp immunostaining on diestrus and KISS1R mRNA levels on proestrus. These changes were accompanied by reduced endometrial glands, reduced uterine proliferative activity, and reduced ERα gene and protein expression. Additionally, hypothyroidism led to reduced uterine gene expression of LIF, BMP2, WNT4, and HAND2. On the other hand, thyroid hypofunction increased uterine PR and TRα immunostaining, while it reduced PGR gene expression on diestrus. These findings demonstrate that hypothyroidism reduces the expression of Kiss1/Kiss1r system in the uterus, which is associated with disrupted uterine estrogen and progesterone signaling and reduced expression of uterine receptivity mediators across the rat estrous cycle.