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The study aimed to examine effects of (-)-epigallocatechin-3-gallate (EGCG) on energy metabolism and mitochondrial dynamics in mouse model of renal injury caused by doxorubicin (DOX). Here, mice were divided into Control group, EGCG-only treated group, DOX group, and three doses of EGCG plus DOX groups. Our results showed that EGCG behaved beneficial effects against kidney injury via attenuation of pathological changes in kidney tissue, which was confirmed by reducing serum creatinine (SCr), blood urea nitrogen (BUN), and apoptosis. Subsequently, changes in reactive oxygen species generation, malondialdehyde content, and activities of antioxidant enzymes were considerably ameliorated in EGCG + DOX groups when compared to DOX group. Furthermore, EGCG-evoked renal protection was associated with increases of mitochondrial membrane potential and decreases of mitochondrial fission protein Dynamin-related protein 1 (Drp1). Moreover, changing glycolysis into mitochondrial oxidative phosphorylation was observed, evidenced by controlling activities of malate dehydrogenase (MDH) and hexokinase (HK) in EGCG + DOX groups when compared to DOX group, indicating that reprogramming energy metabolism was linked to EGCG-induced renal protection in mice. Therefore, EGCG was demonstrated to have a protective effect against kidney injury by reducing oxidative damage, metabolic disorders, and mitochondrial dysfunction, suggesting that EGCG has potential as a feasible strategy to prevent kidney injury.
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AIM: Myocardial injury is a significant cause of death. This study investigated the role and underlying mechanism of interferon-regulatory factor-1 (IRF1) in bevacizumab (BVZ)-induced cardiomyocyte injury. METHODS AND RESULTS: HL-1 cells and C57BL/6 mice receiving BVZ treatment were used to establish in vitro and in vivo models of myocardial injury. The relationship between VEGFA and 14-3-3γ was verified through co-immunoprecipitation and Glutathione S Transferase (GST) pull-down assay. Cell viability and apoptosis were analysed by MTT, propidium iodide (PI) staining and flow cytometry. The release of lactate dehydrogenase (LDH), cardiac troponins T (cTnT), and creatine kinase MB (CK-MB) was measured using the enzyme linked immunosorbent assay. The effects of knocking down IRF1 on BVZ-induced mice were analysed in vivo. IRF1 levels were increased in BVZ-treated HL-1 cells. BVZ treatment induced apoptosis, inhibited cell viability, and promoted the release of LDH, cTnT, and CK-MB. IRF1 silencing suppressed BVZ-induced myocardial injury, whereas IRF1 overexpression had the opposite effect. IRF1 regulated VEGFA expression by binding to its promoter, with the depletion of VEGFA or 14-3-3γ reversing the effects of IRF1 knockdown on the cell viability and apoptosis of BVZ-treated HL-1 cells. 14-3-3γ overexpression promoted cell proliferation, inhibited apoptosis, and reduced the release of LDH, cTnT, and CK-MB, thereby alleviating BVZ-induced HL-1 cell damage. In vivo, IRF1 silencing alleviated BVZ-induced cardiomyocyte injury by regulating the VEGFA/14-3-3γ axis. CONCLUSION: The IRF1-mediated VEGFA/14-3-3γ signalling pathway promotes BVZ-induced myocardial injury. Our study provides evidence for potentially new target genes for the treatment of myocardial injury.
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Cardiotoxicidad , Factor A de Crecimiento Endotelial Vascular , Ratones , Animales , Bevacizumab/farmacología , Ratones Endogámicos C57BL , InterferonesRESUMEN
The viscosity distribution of micellar interiors from the very center to the outer surface is dramatically varied, which has been distinguished in theoretical models, yet it remains highly challenging to quantify this issue experimentally. Herein, a series of fluorophore-substituted surfactants DPAC-Fn (n = 3, 5, 7, 9, 11, 13, and 15) are developed by functionalizing the different alkyl-trimethylammonium bromides with the butterfly motion-based viscosity sensor, N,N'-diphenyl-dihydrodibenzo[a,c]phenazine (DPAC). The immersion depth of DPAC units of DPAC-Fn in cetrimonium bromide (C16TAB) micelles depends on the alkyl chain lengths n. From deep (n = 15) to shallow (n = 3), DPAC-Fn in C16TAB micelles exhibits efficient viscosity-sensitive dynamic multicolor emissions. With external standards for quantification, the viscosity distribution inside a C16TAB micelle with the size of â¼4 nm is changed seriously from high viscosity (â¼190 Pa s) in the core center to low viscosity (â¼1 Pa s) near the outer surface. This work provides a tailored approach for powerful micelle tools to explore the depth-dependent microviscosity of micellar interiors.
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A simple dynamic monitoring strategy for chiral self-assembly is achieved by confining the bent-to-planar evolution observed in N,N'-diphenyl-dihydrodibenzo[a,c]phenazine derivatives (DPAC-R/S-GLD). Besides, this approach provides a facile pathway to fabricate architectures with circularly polarized luminescence (CPL) properties.
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Myocardial ischemia/reperfusion injury (MIRI) is a prevalent condition associated with numerous critical clinical conditions. miR-322 has been implicated in MIRI through poorly understood mechanisms. Our preliminary analysis indicated potential interaction of CREB-binding protein (CBP), a transcriptional coactivator and acetyltransferase, with HIF-1α/ß-catenin, which might regulate miR-322 expression. We, therefore, hypothesized that CBP/HIF-1α/ß-catenin/miR-322 axis might play a role in MIRI. Rat cardiomyocytes subjected to oxygen-glucose deprivation /reperfusion (OGD/R) and Langendorff perfused heart model were used to model MIRI in vitro and in vivo, respectively. We used various techniques such as CCK-8 assay, transferase dUTP nick end labeling staining, western blotting, RT-qPCR, chromatin immunoprecipitation (ChIP), dual-luciferase assay, co-immunoprecipitation (Co-IP), hematoxylin and eosin staining, and TTC staining to assess cell viability, apoptosis, and the levels of CBP, HIF-1α, ß-catenin, miR-322, and acetylation. Our results indicate that OGD/R in cardiomyocytes decreased CBP/HIF-1α/ß-catenin/miR-322 expression, increased cell apoptosis and cytokines, and reduced cell viability. However, overexpression of CBP or miR-322 suppressed OGD/R-induced cell injury, while knockdown of HIF-1α/ß-catenin further exacerbated the damage. HIF-1α/ß-catenin bound to miR-322 promoter to promote its expression, while CBP acetylated HIF-1α/ß-catenin for stabilization. Overexpression of CBP attenuated MIRI in rats by acetylating HIF-1α/ß-catenin to stabilize their expression, resulting in stronger binding of HIF-1α/ß-catenin with the miR-322 promoter and subsequent increased miR-322 levels. Therefore, activating CBP/HIF-1α/ß-catenin/miR-322 signaling may be a potential approach to treat MIRI.
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MicroARNs , Daño por Reperfusión Miocárdica , Animales , Ratas , Apoptosis , beta Catenina/genética , beta Catenina/metabolismo , Proteína de Unión a CREB/genética , Proteína de Unión a CREB/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Daño por Reperfusión Miocárdica/genética , Daño por Reperfusión Miocárdica/metabolismo , Miocitos Cardíacos/metabolismoRESUMEN
A simple strategy is presented for the bifunctional detection of environmental organic vapor and temperature by utilizing H-bond interactions to trap a butterfly-vibration-based fluorophore (DPAC-OH) in a polyurethane (PU) matrix. The method opens up a new path for large-scale environmental inspections and the design of dual-response luminescent materials.
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Myocardial ischemia/reperfusion injury (MIRI) is a major cause of heart failure after myocardial infarction. It has been reported that miR-322 is involved in MIRI progression, while the molecular mechanism of miR-322 in regulating MIRI progression needs to be further probed. MIRI cell model was established by oxygen-glucose deprivation/reoxygenation (OGD/R). Cell viability was assessed using MTS assay. Flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining were employed to analyze cell apoptosis. In addition, the interactions between miR-322, Smad7/Smurf2, hypoxia-inducible factor alpha (HIF-1α), and ß-catenin were verified by dual-luciferase reporter gene assay. Our results displayed that miR-322 was significantly downregulated in OGD/R-treated H9c2 cells, and its overexpression resulted in increased cell viability and reduced the apoptosis. Smurf2 and Smad7 were identified as the direct targets of miR-322. Smad7 knockdown or Smurf2 knockdown increased OGD/R-treated H9c2 cell viability and suppressed the apoptosis. Meanwhile, miR-322 mimics abolished the mitigating effect of Smad7 or Smurf2 overexpression on MIRI. In addition, the Smad3/ß-catenin pathway was identified as the downstream pathway of Smurf2/Smad7. Moreover, it was found that HIF-1α interacted with the miR-322 promoter, and ß-catenin interacted with the HIF-1α promoter to form a loop. HIF-1α-induced upregulated miR-322 activated the Smad3/ß-catenin pathway by targeting Smurf2 and Smad7 to improve MIRI; meanwhile, ß-catenin/HIF-1α formed a positive feedback loop to continuously improve MIRI.
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MicroARNs , Infarto del Miocardio , Daño por Reperfusión Miocárdica , Humanos , Apoptosis , beta Catenina/metabolismo , Retroalimentación , MicroARNs/metabolismo , Infarto del Miocardio/genética , Daño por Reperfusión Miocárdica/metabolismo , Miocitos Cardíacos/metabolismo , Proteína smad3/metabolismo , Proteína smad7/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismoRESUMEN
Pyroelectric infrared (PIR) sensors are low-cost, low-power, and highly reliable sensors that have been widely used in smart environments. Indoor localization systems may be wearable or non-wearable, where the latter are also known as device-free localization systems. Since binary PIR sensors detect only the presence of a subject's motion in their field of view (FOV) without other information about the actual location, information from overlapping FOVs of multiple sensors can be useful for localization. This study introduces the PIRILS (pyroelectric infrared indoor localization system), in which the sensing signal processing algorithms are augmented by deep learning algorithms that are designed based on the operational characteristics of the PIR sensor. Expanding to the detection of multiple targets, the PIRILS develops a quantized scheme that exploits the behavior of an artificial neural network (ANN) model to demonstrate localization performance in tracking multiple targets. To further improve the localization performance, the PIRILS incorporates a data augmentation strategy that enhances the training data diversity of the target's motion. Experimental results indicate system stability, improved positioning accuracy, and expanded applicability, thus providing an improved indoor multi-target localization framework.
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Algoritmos , Inteligencia Artificial , Redes Neurales de la Computación , Procesamiento de Señales Asistido por Computador , Movimiento (Física)RESUMEN
Single-molecule conductance measurements for 9,14-diphenyl-9,14-dihydrodibenzo[a,c]phenazine (DPAC) may offer unique insight into the bent-to-planar photocycle between the ground and excited states. Herein, we employ DPAC derivative DPAC-SMe as the molecular prototype to fabricate single-molecule junctions using the scanning tunneling microscope break junction technique and explore photoconductance dependence on the excited-state structural/electronic changes. We find up to â¼200% conductance enhancement of DPAC-SMe under continuous 340 nm light irradiation than that without irradiation, while photoconductance disappears in the case where structural evolution of the DPAC-SMe is halted through macrocyclization. The in situ conductance modulation as pulsed 340 nm light irradiation is monitored in the DPAC-SMe-based junctions alone, suggesting that the photoconductance of DPAC-SMe stems from photoinduced intramolecular planarization. Theoretical calculations reveal that the photoinduced structural evolution brings about a significant redistribution of the electron cloud density, which leads to the appearance of Fano resonance, resulting in enhanced conductance through the DPAC-SMe-fabricated junctions. This work provides evidence of bent-to-planar photocycle-induced conductance differences at the single-molecule level, offering a tailored approach for tuning the charge transport characteristics of organic photoelectronic devices.
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Electrónica , NanotecnologíaRESUMEN
Pyroelectric Infrared (PIR) sensors are low-cost, low-power, and highly reliable sensors that have been widely used in smart environments. Indoor localization systems can be categorized as wearable and non-wearable systems, where the latter are also known as device-free localization systems. Since the binary PIR sensor detects only the presence of a human motion in its field of view (FOV) without any other information about the actual location, utilizing the information of overlapping FOV of multiple sensors can be useful for localization. In this study, a PIR detector and sensing signal processing algorithms were designed based on the characteristics of the PIR sensor. We applied the designed PIR detector as a sensor node to create a non-wearable cooperative indoor human localization system. To improve the system performance, signal processing algorithms and refinement schemes (i.e., the Kalman filter, a Transferable Belief Model, and a TBM-based hybrid approach (TBM + Kalman filter)) were applied and compared. Experimental results indicated system stability and improved positioning accuracy, thus providing an indoor cooperative localization framework for PIR sensor networks.
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Algoritmos , Procesamiento de Señales Asistido por Computador , Humanos , Movimiento (Física)RESUMEN
Herein, a molecular pixel system for full-color luminescence reproduction is achieved by adjusting the colorless mixtures of two matching fluorophores, i.e., polarity-insensitive 9,14-diphenyl-9,14-dihydrodibenzo[a,c]phenazine (DPAC) as the fixed red primary color and polarity-sensitive dansylamide (DSA) as dynamic blue to green primary colors. DPAC and DSA possess independent emission properties free from electron and energy transfer crosstalk between them because of their close frontier molecular orbitals as well as similar absorptions below 400 nm. According to the additive color theory, under diverse mixing ratios and various polarities, a smooth emission color change is realized in the triangle surrounded by the luminophores in the chromaticity diagram with accurate prediction and expedient reproduction. The principle of this system may open an innovative route for the development of powerful full-color luminescent materials, for example, ratiometric fluorescent polarity sensors and invisible fluorescent crayons.
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Controllable aggregation-induced emission luminogens (AIEgens) by photoexcitation can be conducted within a single solvent, thus opening new opportunities for preparing and processing smart materials. However, undesired side-reactions like photooxidation that can easily occur in the organic phase remain, limiting their applications. To enhance the operability of photoexcitation-controlled AIEgens (to specifically produce a phosphorescence characteristic) in the organic phase, in this work, we employ a typical prototype, hexathiobenzene, usually as the specific phosphorescent group, and investigate a series of physical and chemical factors, such as light intensity, dissolved oxygen content, and solvent polarity, to explore ways to control the photoexcitation-controllable AIEgens against the impurities from side-reactions. An organogel strategy was also developed to minimize interference factors and improve the practical application ability. We believe that the presented results provide new insights into the further development of the photoexcitation-based functional materials and the promotion of their practical usage.
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Long-standing radical species have raised noteworthy concerns in organic functional chemistry and materials. However, there remains a substantial challenge to produce long-standing radicals by light, because of the structural dilemmas between photoproduction and stabilization. Herein, we present a pyrrole and chloride assisted photochromic structure to address this issue. In this well-selected system, production and stabilization of a radical species were simultaneously found accompanied by a photochemical process in chloroform. Theoretical study and mechanism construction indicate that the designed π-system provides a superior spin-delocalization effect and a large steric effect, mostly avoiding possible consumptions and making the radical stable for hours even under an oxygen-saturated condition. Moreover, this radical system can be applied for a visualized and quantitative detection towards peroxides, such as 2,2,6,6-tetramethylpiperidine-1-oxyl, hydrogen peroxide, and ozone. As the detection relies on a radical capturing mechanism, a higher sensing rate was achieved compared to traditional redox techniques for peroxide detection.
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In the development of modern high-performance photoelectric materials, the gated photochromic materials have attracted wide attention. However, the integration of varying signal regulations into gated photochromism to construct efficient photochromic materials is still an urgent necessity. Herein, we designed and synthesized a new gated photoswitching DTEP based on a Schiff base with a diarylethene core. The photochromic properties of compound DTEP can be regulated to different degrees by multiple stimuli, including UV/visible light, Cu2+ and Ni2+. The compound DTEP showed different response abilities to Cu2+ and Ni2+, due to the diverse complexation modes between DTEP and Cu2+ as well as Ni2+. The photochromic properties of compound DTEP could be inhibited completely by the introduction of Cu2+ to form a 1 : 1 complexation, while the weak gated photochromism could be found from the DTEP-Ni2+ complex in a 1 : 2 stoichiometry. Relying on such varying degrees of gated photochromic properties, a new molecular logic circuit was constructed to undertake complicated logical operations.
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Gated photochromism is of interest for the operation and control of modern high-tech optofunctional materials. For further advancing this topic towards the achievement of multifunctional molecular switching, however, it remains a great challenge to incorporate multiple fluorescence regulation into gated photochromism in one unimolecular system. Herein, it is reported that a dithienylethene derivative DTEN with a Schiff base connection can be facilely synthesized by one-step coupling, and it enabled distinct color and spectral changes upon different stimuli, including ultraviolet, visible light, Ni2+ , and Al3+ . Relying on hydrazine and hydroxy units in this molecule, compound DTEN exhibited novel Ni2+ -locked photochromic characteristics originating from complexation of the compound with Ni2+ in a 2:1 stoichiometry. On the other hand, a 1:1 complexation between compound DTEN and Al3+ could allow both of the initial and photostationary states of DTEN to display fluorescent enhancement and a redshift, realizing a dual-fluorescence "turn-on" sensing of Al3+ by light. On this basis, it is argued that the switching of the coordination mode between DTEN and Ni2+ or Al3+ brings up the possibility of tunable photoswitching by multiple stimuli, which offers a novel way for future development of multifunctional switching materials with different input and output signals, as exemplified by the construction of a delicate molecular circuit.
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OBJECTIVES: This study was designed to investigate the effects and the mechanism of Tanshinone IIA (TIIA) on endotoxic shock-induced lung injury in a mouse model. METHODS: Mice were administered intraperitoneally with TIIA (10 mg/kg) 0.5 h before lipopolysaccharide (LPS) challenge and then received additional injections every 24 h during the 3-day experimental period. The physiological indexes, the survival rate and the parameters for lung injury were examined. The protein levels of Sirt1, and the acetylation and activation of NF-κB p65 were determined. The expression and secretion of pro-inflammatory factors were evaluated, respectively. KEY FINDINGS: Treatment with TIIA significantly improved physiological indexes and increased the survival rate of mice in response to LPS challenge. TIIA treatment displayed an obvious up-regulation of Sirt1 protein, in accompany with reduced acetylation and activation of NF-κB p65 following LPS stimulation. In addition, TIIA attenuated LPS-induced lung injury and prevented the expression and secretion of pro-inflammatory factors. However, the protective effects of TIIA were abolished by Sirt1 inhibitor. CONCLUSIONS: Tanshinone IIA prevents LPS-induced secretion of pro-inflammatory cytokines thus exerts protective effects against acute lung injury, probably via modulation of Sirt1/NF-κB signalling pathway.
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Abietanos/farmacología , Lesión Pulmonar Aguda/tratamiento farmacológico , Sirtuina 1/metabolismo , Lesión Pulmonar Aguda/patología , Animales , Carbazoles/farmacología , Citocinas/metabolismo , Lipopolisacáridos , Pulmón/efectos de los fármacos , Pulmón/patología , Ratones , Ratones Endogámicos C57BL , Choque Séptico/tratamiento farmacológico , Transducción de Señal/efectos de los fármacos , Factor de Transcripción ReIA/metabolismoRESUMEN
Myocardial ischemia-reperfusion (I/R) is a common and lethal disease that threatens people's life worldwide. The underlying mechanisms are under intensive study and yet remain unclear. Here, we explored the function of miR-322/503 in myocardial I/R injury. We used isolated rat perfused heart as an in vivo model and H9c2 cells subjected with the oxygen and glucose deprivation followed by reperfusion as in vitro model to study myocardial I/R injury. 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to measure the infarct size, and terminal deoxynucleotidyl transferase dUTP-mediated nick-end label (TUNEL) staining was used to examine apoptosis. Quantitative RT-PCR and Western blot were used to determine expression levels of miR-322/503, Smad ubiquitin regulatory factor 2 (Smurf2), enhancer of zeste homolog 2 (EZH2), p-Akt, and p-GSK3ß. Overexpression of miR-322/503 decreased infarct size, inhibited cell apoptosis, and promoted cell proliferation through upregualtion of p-Akt and p-GSK3ß. Thus the expression of miR-322/503 was reduced during I/R process. On the molecular level, miR-322/503 directly bound Smurf2 mRNA and suppressed its translation. Smurf2 ubiquitinated EZH2 and degraded EZH2, which could activate Akt/GSK3ß signaling. Our study demonstrates that miR-322/503 plays a beneficial role in myocardial I/R injury. By inhibition of Smurf2 translation, miR-322/503 induces EZH2 expression and activates Akt/GSK3ß pathway, thereby protecting cells from ischemia reperfusion injury.
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Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , MicroARNs/metabolismo , Infarto del Miocardio/prevención & control , Daño por Reperfusión Miocárdica/prevención & control , Miocitos Cardíacos/enzimología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Animales , Apoptosis , Sitios de Unión , Hipoxia de la Célula , Línea Celular , Proliferación Celular , Modelos Animales de Enfermedad , Glucosa/deficiencia , Preparación de Corazón Aislado , Masculino , MicroARNs/genética , Infarto del Miocardio/enzimología , Infarto del Miocardio/genética , Infarto del Miocardio/patología , Daño por Reperfusión Miocárdica/enzimología , Daño por Reperfusión Miocárdica/genética , Daño por Reperfusión Miocárdica/patología , Miocitos Cardíacos/patología , Fosforilación , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Sprague-Dawley , Transducción de Señal , Ubiquitina-Proteína Ligasas/genéticaRESUMEN
Cardiotoxicity limits the clinical applications of doxorubicin (Dox), which mechanism might be excess generation of intracellular ROS. Quercetin (Que) is a flavonoid that possesses anti-oxidative activities, exerts myocardial protection. We hypothesized that the cardioprotection against Dox injury of Que involved 14-3-3γ, and mitochondria. To investigate the hypothesis, we treated primary cardiomyocytes with Dox and determined the effects of Que pretreatment with or without 14-3-3γ knockdown. We analyzed various cellular and molecular indexes. Our data showed that Que attenuated Dox-induced toxicity in cardiomyocytes by upregulating 14-3-3γ expression. Que pretreatment increased cell viability, SOD, catalase, and GPx activities, GSH levels, MMP and the GSH/GSSG ratio; decreased LDH and caspase-3 activities, MDA and ROS levels, mPTP opening and the percentage of apoptotic cells. However, Que's cardioprotection were attenuated by knocking down 14-3-3γ expression using pAD/14-3-3γ-shRNA. In conclusion, Que protects cardiomyocytes against Dox injury by suppressing oxidative stress and improving mitochondrial function via 14-3-3γ.
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Proteínas 14-3-3/metabolismo , Antioxidantes/farmacología , Doxorrubicina/toxicidad , Mitocondrias Cardíacas/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Quercetina/farmacología , Proteínas 14-3-3/genética , Animales , Apoptosis/efectos de los fármacos , Células Cultivadas , Técnicas de Silenciamiento del Gen , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias Cardíacas/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Poro de Transición de la Permeabilidad Mitocondrial , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Objectives: In this study, we aim to determine how CUG-expansion and the abundance of Celf1 regulates normal myocyte differentiation and reveal the role ofmiR-206 in myotonic dystrohy and explore a possible gene therapy vector. Methods: we set up CUG-expansion and Celf1 overexpression C2C12 cell models to imitate the myocyte differentiation defects of DM1, then transfected AdvmiR-206 into cell models, tested the level of myogenic markers MyoD, MyoG, Mef2c, Celf1 by RT-PCRand Western Blotting, detected myotube formation by myosin heavy chain immunostaining. Result: 3'-UTR CUG-expansion leads to myotube defects and impaired myoblasts differentiation. Overexpression of Celf1 inhibits myoblast differentiation and impairs differentiation. Knockdown of Celf1 partially rescues differentiation defects of myoblasts harboring CUG-expansion. miR-206 incompletely reverses myoblast differentiation inhibition induced by CUG-expansion and partially recuses myoblast differentiation defects induced by Celf1 overexpression. Conclusions: Ectopic miR-206 mimicking the endogenous temporal patterns specifically drives a myocyte program that boosts myoblast lineages, likely by promoting the expression of MyoD to rectify the myogenic deficiency by stimulating the accumulation of Celf1. Abbreviations: DMPK: (dystrophia myotonica protein kinase); 3'-UTR: (3'-untranslated region); MBNL1: (muscleblind-like [Drosophila]); DM1: (myotonic dystrophy type 1); GFP: (green fluorescent protein); RT-PCR: (quantitative reverse transcriptase-polymerase chain reaction); shRNA: (short hairpin RNA).
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Proteínas CELF1/antagonistas & inhibidores , MicroARNs/metabolismo , Desarrollo de Músculos/fisiología , Mioblastos/metabolismo , Distrofia Miotónica/metabolismo , Animales , Diferenciación Celular/fisiología , Línea Celular , Ratones , Proteína Quinasa de Distrofia Miotónica/genética , Proteína Quinasa de Distrofia Miotónica/metabolismoRESUMEN
Bevacizumab (BVZ) is the first recombinant humanized monoclonal antibody against vascular endothelial growth factor (VEGFA) approved by the FDA for the treatment of different kinds of cancers, especially colorectal cancer. Although the anti-tumor effects have been verified, the side effects of BVZ are also noteworthy, among which, cardiotoxicity may be the most serious side effect of BVZ. However, the exact mechanisms of cardiotoxicity induced by BVZ have been little explored. This study was conducted in vitro in a human cardiac myocyte (HCM) model. MTT assay was conducted to determine BVZ-stimulated cell viability. For testing the function and mechanism, the cells were transfected with miR-140-5p mimics, miR-140-5p inhibitor and/or VEGFA small interfering RNA (si-VEGFA). Then, apoptosis of the cells was detected via annexin V/propidium iodide (AV-PI) staining followed by flow cytometry. qRT-PCR and western blot assays were applied to measure gene expression (i.e. mRNA) and protein levels, respectively. The CK, LDH, SOD, CAT and GSH-Px activities and MDA level were determined using commercial kits. ROS levels were determined by DCFH-DA assay. Mitochondrial membrane potential was measured by JC-1 assay. Dual-luciferase reporter assay was used to detect the interaction between miR-140-5p and VEGFA. BVZ could inhibit HCM proliferation and induce apoptosis. miR-140-5p was upregulated in response to BVZ treatment and miR-140-5p restraint could alleviate HCM damage caused by BVZ treatment. In contrast, VEGFA and 14-3-3γ expressions were down-regulated by BVZ, and miR-140-5p could inhibit the expression of 14-3-3γ by directly targeting VEGFA. Moreover, VEGFA suppression enhanced HCM injury stimulated by BVZ and partially reversed the functional role of the miR-140-5p inhibitor in BVZ-treated cells. Taken together, miR-140-5p promoted BVZ-treated cardiomyocyte toxicity by targeting the VEGFA/14-3-3γ signal pathway. Collectively, miR-140-5p mediated the BVZ-induced cytotoxicity to cardiomyocytes by targeting the VEGFA/14-3-3γ signal pathway, indicating that miR-140-5p may be a novel target for treating BVZ-induced cardiotoxicity.