RESUMO
Hypoadiponectinemia is associated with an increased risk of coronary artery disease. Although adiponectin replenishment mitigates neointimal hyperplasia and atherosclerosis in mouse models, adiponectin therapy has been hampered in a clinical setting due to its large molecular size. Recent studies demonstrate that AdipoRon (a small-molecule adiponectin receptor agonist) improves glycemic control in type 2 diabetic mice and attenuates postischemic cardiac injury in adiponectin-deficient mice, in part, through activation of AMP-activated protein kinase (AMPK). To date, it remains unknown as to whether AdipoRon regulates vascular smooth muscle cell (VSMC) proliferation, which plays a major role in neointima formation. In the present study, oral administration of AdipoRon (50mg/kg) in C57BL/6J mice significantly diminished arterial injury-induced neointima formation by â¼57%. Under in vitro conditions, AdipoRon treatment led to significant inhibition of platelet-derived growth factor (PDGF)-induced VSMC proliferation, DNA synthesis, and cyclin D1 expression. While AdipoRon induced a rapid and sustained activation of AMPK, it also diminished basal and PDGF-induced phosphorylation of mTOR and its downstream targets, including p70S6K/S6 and 4E-BP1. However, siRNA-mediated AMPK downregulation showed persistent inhibition of p70S6K/S6 and 4E-BP1 phosphorylation, indicating AMPK-independent effects for AdipoRon inhibition of mTOR signaling. In addition, AdipoRon treatment resulted in a sustained and transient decrease in PDGF-induced phosphorylation of Akt and ERK, respectively. Furthermore, PDGF receptor-ß tyrosine phosphorylation, which controls the phosphorylation state of Akt and ERK, was diminished upon AdipoRon treatment. Together, the present findings suggest that orally-administered AdipoRon has the potential to limit restenosis after angioplasty by targeting mTOR signaling independent of AMPK activation.
Assuntos
Proliferação de Células/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Neointima/tratamento farmacológico , Piperidinas/uso terapêutico , Fator de Crescimento Derivado de Plaquetas/metabolismo , Receptores de Adiponectina/agonistas , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Artérias/efeitos dos fármacos , Artérias/lesões , Artérias/metabolismo , Artérias/patologia , Linhagem Celular , Humanos , Masculino , Camundongos Endogâmicos C57BL , Músculo Liso Vascular/citologia , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Neointima/metabolismo , Neointima/patologia , Receptores de Adiponectina/metabolismo , Serina-Treonina Quinases TOR/metabolismoRESUMO
BACKGROUND/AIMS: Pioglitazone (PIO), an antidiabetic drug, has been shown to attenuate vascular smooth muscle cell (VSMC) proliferation, which is a major event in atherosclerosis and restenosis after angioplasty. Till date, the likely contributory role of AMP-activated protein kinase (AMPK) toward PIO inhibition of VSMC proliferation has not been examined in vivo. This study is aimed at determining whether pharmacological inhibition of AMPK would prevent the inhibitory effect of PIO on neointima formation in a mouse model of arterial injury. METHODS: Male CJ57BL/6J mice were subjected to femoral artery injury using guidewire. PIO (20 mg/kg/day) was administered orally 1 day before surgery and for 3 weeks until sacrifice in the absence or presence of compound C (an AMPK inhibitor). Injured femoral arteries were used for morphometric analysis of neointima formation. Aortic tissue lysates were used for immunoblot analysis of phosphorylated AMPK. RESULTS: PIO treatment resulted in a significant decrease in intima-to-media ratio by â¼50.3% (p < 0.05, compared with vehicle control; n = 6), which was accompanied by enhanced phosphorylation of AMPK by â¼85% in the vessel wall. Compound C treatment led to a marked reduction in PIO-mediated inhibition of neointima formation. CONCLUSION: PIO attenuates injury-induced neointima formation, in part, through the activation of AMPK.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Hipoglicemiantes/farmacologia , Neointima/prevenção & controle , Tiazolidinedionas/farmacologia , Administração Oral , Animais , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Artéria Femoral/efeitos dos fármacos , Artéria Femoral/lesões , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso Vascular/citologia , Miócitos de Músculo Liso/metabolismo , Fosforilação , PioglitazonaRESUMO
Tumor necrosis factor-α (TNFα) and thrombospondin-1 (TSP-1) are well-known mediators of inflammation. However, a causal relationship between TNFα stimuli and TSP-1 expression in endothelial cell stress, and the underlying mechanisms has not yet been investigated. In our study, human microvascular endothelial cells (hMEC) were treated with TNFα and analyzed for endothelial dysfunction, TSP-1 expression, and associated mechanisms. TNFα treatment induced a dose-dependent increase in TSP-1 expression in hMEC associated with increased endothelial permeability, apoptosis, and reduced proliferation. Whereas TNFα activated Akt, ERK, and P38 mitogen-activated protein kinase (P38 MAPK) simultaneously in hMEC, inhibitors of Akt and P38 MAPK, but not ERK blunted TNFα-induced TSP-1 expression. Silencing of NFκB gene had no significant effect on TNFα-induced TSP-1 expression. Our study demonstrates the novel role of TNFα in inducing inflammatory stress response in hMEC through Akt- and P38 MAPK-mediated expression of TSP-1, independent of NFκB signaling.
Assuntos
Proteínas Proto-Oncogênicas c-akt/metabolismo , Trombospondina 1/metabolismo , Fator de Necrose Tumoral alfa/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Apoptose , Permeabilidade Capilar , Proliferação de Células , Sobrevivência Celular , Células Endoteliais , Endotélio Vascular , Expressão Gênica , Humanos , Sistema de Sinalização das MAP Quinases , Microvasos , Fosforilação , Processamento de Proteína Pós-Traducional , Estresse Fisiológico/imunologia , Trombospondina 1/genética , Ativação TranscricionalRESUMO
AIMS/HYPOTHESIS: Obesity and hypertension, known pro-inflammatory states, are identified determinants for increased retinal microvascular abnormalities. However, the molecular link between inflammation and microvascular degeneration remains elusive. Thioredoxin-interacting protein (TXNIP) is recognised as an activator of the NOD-like receptor pyrin domain containing-3 (NLRP3) inflammasome. This study aims to examine TXNIP expression and elucidate its role in endothelial inflammasome activation and retinal lesions. METHODS: Spontaneously hypertensive (SHR) and control Wistar (W) rats were compared with groups fed a high-fat diet (HFD) (W+F and SHR+F) for 8-10 weeks. RESULTS: Compared with W controls, HFD alone or in combination with hypertension significantly induced formation of acellular capillaries, a hallmark of retinal ischaemic lesions. These effects were accompanied by significant increases in lipid peroxidation, nitrotyrosine and expression of TXNIP, nuclear factor κB, TNF-α and IL-1ß. HFD significantly increased interaction of TXNIP-NLRP3 and expression of cleaved caspase-1 and cleaved IL-1ß. Immunolocalisation studies identified TXNIP expression within astrocytes and Müller cells surrounding retinal endothelial cells. To model HFD in vitro, human retinal endothelial (HRE) cells were stimulated with 400 µmol/l palmitate coupled to BSA (Pal-BSA). Pal-BSA triggered expression of TXNIP and its interaction with NLRP3, resulting in activation of caspase-1 and IL-1ß in HRE cells. Silencing Txnip expression in HRE cells abolished Pal-BSA-mediated cleaved IL-1ß release into medium and cell death, evident by decreases in cleaved caspase-3 expression and the proportion of live to dead cells. CONCLUSIONS/INTERPRETATION: These findings provide the first evidence for enhanced TXNIP expression in hypertension and HFD-induced retinal oxidative/inflammatory response and suggest that TXNIP is required for HFD-mediated activation of the NLRP3 inflammasome and the release of IL-1ß in endothelial cells.
Assuntos
Proteínas de Transporte/metabolismo , Inibidores de Caspase/metabolismo , Células Endoteliais/metabolismo , Oftalmopatias/metabolismo , Inflamassomos/metabolismo , Obesidade/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Retina/metabolismo , Animais , Proteínas de Transporte/biossíntese , Proteínas de Ciclo Celular , Morte Celular , Dieta Hiperlipídica , Oftalmopatias/patologia , Inflamação/metabolismo , Masculino , Microcirculação , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR , Obesidade/complicações , Estresse Oxidativo , Ratos , Ratos Wistar , Retina/patologiaRESUMO
BACKGROUND: The learning performance and overall health of students might be impacted by excessive academic stress. While the right amount of stress can help improve learning and performance, too much stress can harm one's mental and physical health as well as academic performance. This research aims to assess the prevalence and use of the beta-blocker propranolol among healthcare students in Saudi Arabia. METHODS: A cross-sectional study was conducted among healthcare students at Umm Al-Qura University in Makkah, Saudi Arabia. The participants were sent an electronic questionnaire at random over three months, from June 10 to September 10, 2023. The data were analyzed using RStudio (version 4.2.2), and the categorized data were presented in frequencies and percentages. Fisher's exact test was used to determine the factors associated with propranolol use. The results were reported as odds ratios (OR) and 95% confidence intervals (CI). A p-value of < 0.05 was considered statistically significant. RESULTS: The study comprised 582 participants, of whom (51.7%, n=301) fell within the age range of 24 to 26 years, (63.1%, n=367) were male, and (59.3%, n=345) were enrolled in the College of Medicine and Surgery. The majority of respondents (73.7%, n=28) reported that educational materials such as medical books were their primary source of information regarding the impact of beta-blockers on anxiety. Among those who used propranolol, over two-thirds (68.4%, n=26) had taken it before the Objective Structured Clinical Examination (OSCE). About a quarter of the participants (26.1%, n=151) believed that propranolol was being misused by healthcare students, and (21.3%, n=123) believed that the drug could enhance academic performance. CONCLUSION: The primary motives for taking propranolol were to alleviate anxiety before OSCEs and enhance performance during presentations. The participants showed some understanding of the impact of propranolol. Nevertheless, it is imperative to impart knowledge to them about the potential hazards linked to the misuse of beta-blockers.
RESUMO
Objectives: Worldwide, diabetic neuropathy (DN) is a major complication of diabetes mellitus. The direct renin inhibitor aliskiren is recognized as a treatment for cardiovascular disease in diabetic patients, but little is known about its potential benefits in cases of diabetic neuropathy. Accordingly, we investigated the effects of aliskiren (ALIS) and gliclazide (GLZ) and their combination therapy on peripheral neuropathy in streptozotocin-induced diabetic rats. Methods: In total, 112 adult Sprague-Dawley rats were used for this study. Diabetes was induced using streptozotocin (STZ), whereas the control group was treated with an equal volume of citrate buffer. The diabetic rats were divided randomly into six groups according to the proposed treatment regime: diabetic control (DC), gliclazide (GLZ), aliskiren (ALIS), ramipril (RAM), (GLZ + ALIS) and (GLZ + RAM). Behavioural responses to thermal (hot-plate) and mechanical (tail-pinch) pain were evaluated. After eight weeks of daily treatments, the animals were fasted and sacrificed. The blood samples were collected, with the serum separated and subjected to various biochemical and enzyme analyses so as to assess the effect of the treatments on diabetic peripheral neuropathy. Results: After 8 weeks, aliskiren alone and in combination with gliclazide therapy had a significant effect (P < .001) in reducing blood glucose levels and showed increased hot-plate and tail-flick latencies compared with the diabetic control group. The threshold of mechanical hyperalgesia was also significantly elevated (P < .001). Conclusions/Interpretations: These data suggest that either aliskerin alone or in combination with gliclazide can protect against the development and progression of diabetic neuropathy.
Assuntos
Amidas/administração & dosagem , Diabetes Mellitus Experimental/complicações , Neuropatias Diabéticas/etiologia , Neuropatias Diabéticas/prevenção & controle , Fumaratos/administração & dosagem , Neuralgia/etiologia , Neuralgia/prevenção & controle , Animais , Progressão da Doença , Quimioterapia Combinada , Gliclazida/administração & dosagem , Masculino , Ramipril/administração & dosagem , Ratos Sprague-Dawley , Estreptozocina , Resultado do TratamentoRESUMO
Hypoxia is known to promote vasodilation of coronary vessels through several mediators including cardiac-derived adenosine and endothelium-derived prostanoids and nitric oxide. To date, the impact of endogenous glycogen depletion in vascular smooth muscle and the resultant alterations in cellular energy state (e.g., AMP-activated protein kinase, AMPK) on the contractile response to G protein-coupled receptor agonists (e.g., serotonin, 5-HT) has not yet been studied. In the present study, ex vivo exposure of endothelium-denuded human saphenous vein rings to hypoxic and glucose-deprived conditions during KCl-induced contractions for 30 min resulted in a marked depletion of endogenous glycogen by â¼80% (from â¼1.78 µmol/g under normoxia to â¼0.36 µmol/g under hypoxia). Importantly, glycogen-depleted HSV rings, which were maintained under hypoxia/reoxygenation and glucose-deprived conditions, exhibited significant increases in basal AMPK phosphorylation (â¼6-fold ↑) and 5-HT-induced AMPK phosphorylation (â¼19-fold ↑) with an accompanying suppression of 5-HT-induced maximal contractile response (â¼68% ↓), compared with respective controls. Exposure of glycogen-depleted HSV rings to exogenous D-glucose, but not the inactive glucose analogs, prevented the exaggerated increase in 5-HT-induced AMPK phosphorylation and restored 5-HT-induced maximal contractile response. In addition, the ability of exogenous D-glucose to rescue cellular stress and impaired contractile function occurred through GLUT1-mediated but insulin/GLUT4-independent mechanisms. Together, the present findings from clinically-relevant human saphenous vein suggest that the loss of endogenous glycogen in vascular smooth muscle and the resultant accentuation of AMPK phosphorylation by GPCR agonists may constitute a yet another mechanism of metabolic vasodilation of coronary vessels in ischemic heart disease.