RESUMO
Myocardial infarction (MI) is the leading cause of death worldwide, and oxidative stress is part of the process that causes MI. Calycosin, a naturally occurring substance with cardioprotective properties, is one of the major active constituents in Radix Astragali. In this study, effect of Calycosin was investigated in vivo and in vitro to determine whether it could alleviate oxidative stress and oxidative stress-induced cardiac apoptosis in neonatal cardiomyocytes (NCMs) via activation of aldehyde dehydrogenase 2 (ALDH2). Calycosin protected against oxidative stress and oxidative stress-induced apoptosis in NCMs. Molecular docking revealed that the ALDH2-Calycosin complex had a binding energy of -9.885 kcal/mol. In addition, molecular docking simulations demonstrated that the ALDH2-Calycosin complex was stable. Using BLI assays, we confirmed that Calycosin could interact with ALDH2 (KD = 1.9 × 10-4 M). Furthermore, an ALDH2 kinase activity test revealed that Calycosin increased ALDH2 activity, exhibiting an EC50 of 91.79 µM. Pre-incubation with ALDH2 inhibitor (CVT-10216 or disulfiram) reduced the cardio-protective properties Calycosin. In mice with MI, Calycosin therapy substantially reduced myocardial apoptosis, oxidative stress, and activated ALDH2. Collectively, our findings clearly suggest that Calycosin reduces oxidative stress and oxidative stress-induced apoptosis via the regulation of ALDH2 signaling, which supports potential therapeutic use in MI.
Assuntos
Infarto do Miocárdio , Miócitos Cardíacos , Camundongos , Animais , Aldeído-Desidrogenase Mitocondrial/metabolismo , Simulação de Acoplamento Molecular , Estresse Oxidativo , Apoptose , Aldeído Desidrogenase/metabolismoRESUMO
PURPOSE: We present a retrospective study of patients with multilevel contiguous tuberculous spondylitis of thoracic region that underwent single-stage posterolateral debridement and fusion and following posterior instrumentation. METHODS: From June 2000 to March 2009, 870 consecutive spinal tubercular patients including 36 patients who were diagnosed and treated as multilevel contiguous thoracic spinal tuberculosis in our institution. Apart from five patients being treated conservatively, the 31 cases received surgery by single-stage posterolateral debridement, fusion, following posterior instrumentation and postural drainage. The patients were evaluated based on the Frankel scoring system, kyphotic Cobb angle, and visual analog scale (VAS) pain score. RESULTS: The mean duration of postoperative follow-up was 79.2 ± 9.9 months (range 62-98 months). Neither mortalities nor any major complications were found. Solid bony fusion was achieved in all patients. No patients with neurological deficit deteriorated postoperatively. According to Frankel scoring system, 7 cases were rated as Grade D, 24 cases as Grade E at last follow-up. The average preoperative Cobb's angle was 32° (range 21°-39°). The average early postoperative Cobb's angle was 23° (range 15°-32°). The mean latest postoperative Cobb's angle was 26° (range 20°-32°), with a small loss of correction at last follow-up. Pre-op VAS was 8.8 ± 0.7 (range 7-10) and final follow-up was 1.8 ± 1.1. There was a significant difference of VAS between preoperation and the final follow-up. CONCLUSIONS: One-stage surgical treatment for multilevel contiguous spinal tuberculosis by posterolateral debridement, fusion, posterior instrumentation can be an effective and feasible treatment method.
Assuntos
Desbridamento/métodos , Drenagem Postural/métodos , Fusão Vertebral/métodos , Vértebras Torácicas/cirurgia , Tuberculose da Coluna Vertebral/cirurgia , Adulto , Idoso , Desbridamento/efeitos adversos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Medição da Dor , Período Pós-Operatório , Estudos Retrospectivos , Fusão Vertebral/efeitos adversos , Vértebras Torácicas/patologia , Resultado do TratamentoRESUMO
OBJECTIVE: The purpose of this study was to evaluate the outcomes of computed tomography (CT) guidance using percutaneous catheter with low-dose drainage local chemotherapy (modified PCD) and antituberculous therapy (ATT) for the treatment of spinal tuberculosis in children. METHODS: Twenty-seven children suffering from spinal tuberculosis were treated with modified PCD and ATT in our institute from 2002 to 2012. We describe our treatment, which involves CT-guided percutaneous puncture and local chemotherapy (continuous low-dose (20 mL) irrigation). The patients were evaluated based on the Frankel scoring system, the kyphotic Cobb angle, and the erythrocyte sedimentation rate (ESR). RESULTS: All patients were followed up for an average of 31.00 ± 13.94 months. No sinus formation was detected. All patients responded well to this treatment. The ESR values were decreased to normal at last follow-up. The neurological functions show significant improvement after operation. Preoperatively, the kyphotic angle was 22.89 ± 7.06°, and it was measured as 21.19 ± 8.73° at the last visit. CONCLUSIONS: Our results showed that percutaneous intubation and low-dose irrigation under CT guidance (modified percutaneous catheter drainage (MPCD)) and ATT are easy, safe, efficient, and less invasive methods for the treatment of spinal tuberculosis in children.
Assuntos
Abscesso , Antituberculosos/uso terapêutico , Cateterismo/métodos , Tuberculose da Coluna Vertebral , Abscesso/tratamento farmacológico , Abscesso/etiologia , Abscesso/cirurgia , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Masculino , Estudos Retrospectivos , Tomógrafos Computadorizados , Tuberculose da Coluna Vertebral/complicações , Tuberculose da Coluna Vertebral/tratamento farmacológico , Tuberculose da Coluna Vertebral/cirurgiaRESUMO
BACKGROUND: Endothelial dysfunction (ED), characterized by markedly reduced nitric oxide (NO) bioavailability, vasoconstriction, and a shift toward a proinflammatory and prothrombotic state, is an important contributor to hypertension, atherosclerosis, and other cardiovascular diseases. Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) is widely involved in cardiovascular development. Przewaquinone A (PA), a lipophilic diterpene quinone extracted from Salvia przewalskii Maxim, inhibits vascular contraction. PURPOSE: Herein, the goal was to explore the protective effect of PA on ED in vivo and in vitro, as well as the underlying mechanisms. METHODS: A human umbilical vein endothelial cell (HUVEC) model of ED induced by angiotensin II (AngII) was used for in vitro observations. Levels of AMPK, endothelial nitric oxide synthase (eNOS), vascular cell adhesion molecule-1 (VCAM-1), nitric oxide (NO), and endothelin-1 (ET-1) were detected by western blotting and ELISA. A mouse model of hypertension was established by continuous infusion of AngII (1000 ng/kg/min) for 4 weeks using osmotic pumps. Following PA and/or valsartan administration, NO and ET-1 levels were measured. The levels of AMPK signaling-related proteins in the thoracic aorta were evaluated by immunohistochemistry. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) were measured using the tail cuff method. Isolated aortic vascular tone measurements were used to evaluate the vasodilatory function in mice. Molecular docking, molecular dynamics, and surface plasmon resonance imaging (SPRi) were used to confirm AMPK and PA interactions. RESULTS: PA inhibited AngII-induced vasoconstriction and vascular adhesion as well as activated AMPK signaling in a dose-dependent manner. Moreover, PA markedly suppressed blood pressure, activated vasodilation in mice following AngII stimulation, and promoted the activation of AMPK signaling. Furthermore, molecular simulations and SPRi revealed that PA directly targeted AMPK. AMPK inhibition partly abolished the protective effects of PA against endothelial dysfunction. CONCLUSION: PA activates AMPK and ameliorates endothelial dysfunction during hypertension.
Assuntos
Proteínas Quinases Ativadas por AMP , Angiotensina II , Endotélio Vascular , Células Endoteliais da Veia Umbilical Humana , Hipertensão , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo III , Óxido Nítrico , Angiotensina II/farmacologia , Animais , Humanos , Proteínas Quinases Ativadas por AMP/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Masculino , Óxido Nítrico Sintase Tipo III/metabolismo , Hipertensão/tratamento farmacológico , Endotélio Vascular/efeitos dos fármacos , Óxido Nítrico/metabolismo , Camundongos , Salvia/química , Endotelina-1/metabolismo , Molécula 1 de Adesão de Célula Vascular/metabolismo , Quinonas/farmacologia , Simulação de Acoplamento Molecular , Pressão Sanguínea/efeitos dos fármacos , Modelos Animais de DoençasRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Radix Glycyrrhizae (GL), a herbal medicine that is widely available, has shown advantages for a variety of inflammatory diseases. Toll like receptor 4 (TLR4) pathway has been shown to play a key role in the progression of inflammation. AIM OF THE STUDY: The purpose of this study was to investigate the involvement of TLR4 in the anti-inflammatory mechanism of GL extract and its active constituent on acute lung injury (ALI). MATERIALS AND METHODS: A model of inflammation produced by lipopolysaccharide (LPS) was established in C57BL/6 mice and macrophages derived from THP-1. To screen the active components of GL, molecular docking was used. Molecular dynamics and surface plasmon resonance imaging (SPRi) were used to study the interaction of a specific drug with the TLR4-MD2 complex. TLR4 was overexpressed by adenovirus to confirm TLR4 involvement in the anti-inflammatory activities of GL and the chosen chemical. RESULTS: We observed that GL extract significantly reduced both LPS-induced ALI and the production of pro-inflammatory factors including TNF-α, IL-6 and IL-1ß. Additionally, GL inhibited the binding of Alexa 488-labeled LPS (LPS-488) to the membrane of THP-1 derived macrophages. GL drastically reduce on the expression of TLR4 and the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor-B (NF-κB). Furthermore, molecular docking revealed that Licochalcone A (LicoA) docked into the LPS binding site of TLR4-MD2 complex. MD2-LicoA binding conformation was found to be stable using molecular dynamic simulations. SPRi indicated that LicoA bound to TLR4-MD2 recombinant protein with a KD of 3.87 × 10-7 M. LicoA dose-dependently reduced LPS-488 binding to the cell membrane. LicoA was found to significantly inhibit LPS-induced lung damage and inflammation. Furthermore, LicoA inhibited TLR4 expression, MAPK and NF-κB activation in a dose-dependent manner. The inhibitory effects of GL and LicoA on LPS-induced inflammation and TLR4 signaling activation were partly eliminated by TLR4 overexpression. CONCLUSION: Our findings imply that GL and LicoA exert inhibitory effects on inflammation by targeting the TLR4 directly.
Assuntos
Lesão Pulmonar Aguda , Receptor 4 Toll-Like , Camundongos , Animais , Receptor 4 Toll-Like/metabolismo , Lipopolissacarídeos/toxicidade , NF-kappa B/metabolismo , Simulação de Acoplamento Molecular , Camundongos Endogâmicos C57BL , Antígeno 96 de Linfócito/metabolismo , Anti-Inflamatórios/efeitos adversos , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/tratamento farmacológico , Lesão Pulmonar Aguda/metabolismo , Inflamação/induzido quimicamenteRESUMO
BACKGROUND: With an increasing number of myocardial infarction (MI) patients, myocardial fibrosis is becoming a widespread health concern. It's becoming more and more urgent to conduct additional research and investigations into efficient treatments. Ethyl ferulate (EF) is a naturally occurring substance with cardioprotective properties. However, the extent of its impact and the underlying mechanism of its treatment for myocardial fibrosis after MI remain unknown. PURPOSE: The goal of this study was to look into how EF affected the signaling of the TGF-receptor 1 (TGFBR1) in myocardial fibrosis after MI. METHODS: Echocardiography, hematoxylin-eosin (HE) and Masson trichrome staining were employed to assess the impact of EF on heart structure and function in MI-affected mice in vivo. Cell proliferation assay (MTS), 5-Ethynyl-2'-deoxyuridine (EdU), and western blot techniques were employed to examine the influence of EF on native cardiac fibroblast (CFs) proliferation and collagen deposition. Molecular simulation and surface plasmon resonance imaging (SPRi) were utilized to explore TGFBR1 and EF interaction. Cardiac-specific Tgfbr1 knockout mice (Tgfbr1ΔMCK) were utilized to testify to the impact of EF. RESULTS: In vivo experiments revealed that EF alleviated myocardial fibrosis, improved cardiac dysfunction after MI and downregulated the TGFBR1 signaling in a dose-dependent manner. Moreover, in vitro experiments revealed that EF significantly inhibited CFs proliferation, collagen deposition and TGFBR1 signaling followed by TGF-ß1 stimulation. More specifically, molecular simulation, molecular dynamics, and SPRi collectively showed that EF directly targeted TGFBR1. Lastly, knocking down of Tgfbr1 partially reversed the inhibitory activity of EF on myocardial fibrosis in MI mice. CONCLUSION: EF attenuated myocardial fibrosis post-MI by directly suppressing TGFBR1 and its downstream signaling pathway.