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Background and Aims: The role of platelet autophagy in cirrhotic thrombocytopenia (CTP) remains unclear. This study aimed to investigate the impact of platelet autophagy in CTP and elucidate the regulatory mechanism of hydrogen sulfide (H2S) on platelet autophagy. Methods: Platelets from 56 cirrhotic patients and 56 healthy individuals were isolated for in vitro analyses. Autophagy markers (ATG7, BECN1, LC3, and SQSTM1) were quantified using enzyme-linked immunosorbent assay, while autophagosomes were visualized through electron microscopy. Western blotting was used to assess the autophagy-related proteins and the PDGFR/PI3K/Akt/mTOR pathway following treatment with NaHS (an H2S donor), hydroxocobalamin (an H2S scavenger), or AG 1295 (a selective PDGFR-α inhibitor). A carbon tetrachloride-induced cirrhotic BALB/c mouse model was established. Cirrhotic mice with thrombocytopenia were randomly treated with normal saline, NaHS, or hydroxocobalamin for 15 days. Changes in platelet count and aggregation rate were observed every three days. Results: Cirrhotic patients with thrombocytopenia exhibited significantly decreased platelet autophagy markers and endogenous H2S levels, alongside increased platelet aggregation, compared to healthy controls. In vitro, NaHS treatment of platelets from severe CTP patients elevated LC3-II levels, reduced SQSTM1 levels, and decreased platelet aggregation in a dose-dependent manner. H2S treatment inhibited PDGFR, PI3K, Akt, and mTOR phosphorylation. In vivo, NaHS significantly increased LC3-II and decreased SQSTM1 expressions in platelets of cirrhotic mice, reducing platelet aggregation without affecting the platelet count. Conclusions: Diminished platelet autophagy potentially contributes to thrombocytopenia in cirrhotic patients. H2S modulates platelet autophagy and functions possibly via the PDGFR-α/PI3K/Akt/mTOR signaling pathway.
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Soil sickness a severe problem in tobacco production, leading to soil-borne diseases and reduce in tobacco yield. This occurs as a result of the interaction between root exudates and rhizosphere microorganisms, which is however, little studied until now. By combining the field investigation and pot experiment, we found the output yield consistently decreased during the first 10 years of continuous cropping in a tobacco field, but increased at the 15th year (15Y). The root exudate and rhizosphere bacterial community was further analyzed to reveal the underlying mechanism of the suppressive soil formation. Root exudate of 15Y tobacco enriched in amino acids and derivatives, while depleted in the typical autotoxins including phenolic acids and alkaloids. This was correlated to the low microbial diversity in 15Y, but also the changes in community composition and topological properties of the co-occurrence network. Especially, the reduced autotoxins were associated with low Actinobacteria abundance, low network complexity and high network modularity, which significantly correlated with the recovered output yield in 15Y. This study revealed the coevolution of rhizosphere microbiota and root exudate as the soil domesticated by continuous cropping of tobacco, and indicated a potential role of the autotoxins and theirs effect on the microbial community in the formation of suppressive soil.
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Microbiota , Nicotiana , Raíces de Plantas , Rizosfera , Microbiología del Suelo , Nicotiana/microbiología , Nicotiana/crecimiento & desarrollo , Raíces de Plantas/microbiología , Raíces de Plantas/crecimiento & desarrollo , Exudados de Plantas/metabolismo , Suelo/químicaRESUMEN
AIM: Aberrant expression of ATPase sarcoplasmic/endoplasmic retic Ca2+ transporting 2 (ATP2A2) has attracted attention for its pathophysiologic role in pulmonary hypertension (PH). Several miRNAs, including miR-210-5p, have also been reported to be pathogenic factors in PH, but their exact mechanisms remain unknown. This study aimed to elucidate the potential mechanisms of miR-210-5p and ATP2A2 in MCT-induced PH. METHODS: Eighteen Sprague-Dawley rats were randomly divided into two groups-monoclonal (MCT) group and control group-and then administered MCT (60 mg/kg) and saline, respectively. mPAP, PVR, RVHI, WT%, and WA% were significantly increased in PH rats after 3 weeks, confirming that the modeling of PH rats was successful. Subsequently, we determined the expression of ATP2A2 and miR-210-5p in lung tissues using WB and qRT-PCR methods. We established an in vitro model using BMP4 and TGF-ß1 treatment of pulmonary artery smooth muscle cells (PASMCs) and examined the expression of ATP2A2 and miR-210-5p using the same method. To further elucidate the regulatory relationship between ATP2A2 and miR-210-5p, we altered the expression level of miR-210-5p and detected the corresponding changes in ATP2A2 levels. In addition, we demonstrated the relationship by dual luciferase experiments. Finally, the effect of silencing ATP2A2 could be confirmed by the level of cell membrane Ca2+ in PAMSCs. RESULTS: Up-regulation of miR-210-5p and down-regulation of ATP2A2 were observed in the MCT group compared with the control group, which was confirmed in the in vitro model. In addition, elevated miR-210-5p expression decreased the level of ATP2A2 while increasing the proliferation of PASMCs, and the results of the dual luciferase assay further confirmed that ATP2A2 is a downstream target of miR-210-5p. Additionally, silencing ATP2A2 resulted in increased cytoplasmic Ca2+ levels in PAMSCs. CONCLUSION: In MCT-induced PH, miR-210-5p promotes pulmonary vascular remodeling by inhibiting ATP2A2.
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Ferroptosis was reported to be involved in cerebral ischemia-reperfusion injury (CIRI), on which the effects of berberine (BBR) remain unclear. Moreover, based on the critical role of gut microbiota in pleiotropic actions of BBR, we hypothesized that BBR can suppress CIRI-induced ferroptosis by modulating the gut microbiota. In this study, the results showed that BBR obviously attenuated the behavioral deficits of CIRI mice, accompanied with the improved survival rate and neuron damages, as phenocopied by dirty cage experiment. The typical morphological changes in ferroptotic cells and biomarkers of ferroptosis were attenuated in BBR- and its fecal microbiota-treated mice, accompanied by reduced malondialdehyde and reactive oxygen species, and the increased glutathione (GSH). BBR was found to alter the gut microbiota of CIRI mice with decreased abundance of Muribaculaceae, Erysipelotrichaceae, Helicobacteraceae, Streptococcaceae and Tannerellaceae, but elevated Bacteroidaceae and Enterobacteriaceae. KEGG analysis based on the 16S rRNA results indicated that multiple metabolic pathways including ferroptosis and GSH metabolism, were altered by BBR. Oppositely, the antibiotics administration counteracted the protective properties of BBR. Summarily, this study revealed the therapeutic potential of BBR on CIRI via inhibiting neuronal ferroptosis, in which upregulated glutathione peroxidase 1 (GPX1) was possibly involved. Moreover, the BBR-modulated gut microbiota was shown to play the critical role in the underlying mechanism.
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Berberina , Ferroptosis , Microbioma Gastrointestinal , Ratones , Animales , Berberina/farmacología , Berberina/uso terapéutico , ARN Ribosómico 16S , Isquemia/tratamiento farmacológico , ReperfusiónRESUMEN
BACKGROUND: Ethanol-induced gastric mucosal lesions (EGML) is one of the most common digestive disorders for which current therapies have limited outcomes in clinical practice. Prevotella histicola (P. histicola) has shown probiotic efficacy against arthritis, multiple sclerosis and oestrogen deficiency-induced depression in mice; however, its role in EGML remains unclear in spite of its extensive colonisation of the stomach. Ferroptosis, which is characterised by lipid peroxidation, may be involved in EGML. Herein, we aimed to investigate the effects and underlying mechanism of action of P. histicola on EGML in the ferroptosis-dependent pathway. METHODS: P. histicola was intragastrically administered for a week, and deferoxamine (DFO), a ferroptosis inhibitor, was intraperitoneally injected prior to oral ethanol administration. The gastric mucosal lesions and ferroptosis were assessed via histopathological examinations, quantitative real-time PCR, Western blot, immunohistochemistry and immunofluorescence. RESULTS: P. histicola was originally found to attenuate EGML by reducing histopathological changes and lipid reactive oxygen species (ROS) accumulation. The pro-ferroptotic genes of Transferrin Receptor (TFR1), Solute Carrier Family 39 Member 14 (SLC39A14), Haem Oxygenase-1 (HMOX-1), Acyl-CoA Synthetase Long-chain Family Member 4 (ACSL4), Cyclooxygenase 2 (COX-2) and mitochondrial Voltage-dependent Anion Channels (VDACs) were up-regulated; the anti-ferroptotic System Xc-/Glutathione Peroxidase 4 (GPX4) axis was inhibited after ethanol administration. However, the changes of histopathology and ferroptosis-related parameters induced by ethanol were reversed by DFO. Furthermore, P. histicola treatment significantly downregulated the expression of ACSL4, HMOX-1 and COX-2, as well as TFR1 and SLC39A14, on mRNA or the protein level, while activating the System Xc-/GPX4 axis. CONCLUSIONS: We found that P. histicola reduces ferroptosis to attenuate EGML by inhibiting the ACSL4- and VDAC-dependent pro-ferroptotic pathways and activating the anti-ferroptotic System Xc-/GPX4 axis.
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Proteínas de Transporte de Catión , Ferroptosis , Animales , Ratones , Ciclooxigenasa 2 , Administración Oral , EtanolRESUMEN
OBJECTIVE: To develop prediction models for sarcopenia in older patients with hip fracture based on a specific set of serum biomarkers aimed at estimating appendicular skeletal muscle mass and diagnosing sarcopenia. METHODS: Older patients with hip fracture admitted to the First Affiliated Hospital of Wenzhou Medical University from January 2020 to June 2021 were recruited, screened for sarcopenia and tested for peripheral blood levels of specific serum biomarkers preoperatively. Participants were randomly divided into a training set and test set. Common factors were extracted from selected biomarkers through factor analysis, and regression models were established in the training set and verified in the test set. RESULTS: A total of 212 patients were enrolled, and the prevalence of sarcopenia was 22.8% in men and 19.5% in women. Significant differences in cystatin C, estimated glomerular filtration rate based on cystatin C, sarcopenia index, new sarcopenia index, haemoglobin and albumin were observed between patients with and without sarcopenia. Two regression models were developed in the training set. The validation of the test set confirmed that the linear regression model showed good consistency in predicting appendicular skeletal muscle mass index, while the logistic regression model showed high accuracy in predicting sarcopenia. CONCLUSIONS: Both prediction models exhibited potential clinical application value for estimating appendicular skeletal muscle mass and predicting sarcopenia in older patients with hip fracture, providing new insights into the serological diagnosis of sarcopenia.
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Fracturas de Cadera , Sarcopenia , Masculino , Humanos , Femenino , Anciano , Sarcopenia/diagnóstico , Sarcopenia/epidemiología , Cistatina C , Músculo Esquelético , Fracturas de Cadera/diagnóstico , BiomarcadoresRESUMEN
Macrophages, mainly divided into M1 pro-inflammatory and M2 anti-inflammatory types, play a key role in the transition from inflammation to repair after trauma. In chronic inflammation, such as diabetes and complex bone injury, or the process of certain inflammatory specific emergencies, the ratio of M1/M2 cell populations is imbalanced so that M1-macrophages cannot be converted into M2 macrophages in time, resulting in delayed trauma repair. Early and timely transformation of macrophages from the pro-inflammatory M1-type into the pro-reparative M2-type is an effective strategy to guide trauma repair and establish the original homeostasis. We prepared purified nano-platelet vesicles (NPVs) and assessed their effects on macrophage phenotype switching through transcriptome analysis. The results elucidate that NPVs promote pathways related to angiogenesis, collagen synthesis, cell adhesion, and migration in macrophages, and we speculate that these advantages may promote healing in traumatic diseases.
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Inflamación , Macrófagos , Plaquetas , Humanos , Inflamación/metabolismo , Macrófagos/metabolismo , Transducción de Señal , Factor de Crecimiento Transformador betaRESUMEN
BACKGROUND AND AIMS: There is accumulating evidence that gut microbiota plays a key role in cardiovascular diseases. Gut bacteria can transform dietary choline, l-carnitine, and trimethylamine N-oxide (TMAO) into trimethylamine, which can be oxidized into TMAO again in the liver. However, the alterations of the gut microbiota in large artery atherosclerotic (LAA) stroke and cardioembolic (CE) stroke have been less studied. METHODS AND RESULTS: We performed a case-control study in patients with LAA and CE types of strokes. We profiled the gut microbiome using Illumina sequencing of the 16S ribosomal RNA gene (V4-V5 regions), and TMAO was determined via liquid chromatography-tandem mass spectrometry. Our results showed that the TMAO levels in the plasma of patients with LAA and CE strokes were significantly higher than those in controls (LAA stroke, 2931 ± 456.4 ng/mL; CE stroke, 4220 ± 577.6 ng/mL; healthy control, 1663 ± 117.8 ng/mL; adjusted p < 0.05). The TMAO level in the plasma of patients with LAA stroke was positively correlated with the carotid plaque area (rho = 0.333, 95% CI = 0.08-0.55, p = 0.0093). Notably, the composition and the function of gut microbiota in the LAA stroke group were significantly different from those in the control group (FDR-adjusted p-value < 0.05). There was no significant association between gut microbiota and CE stroke in our study. CONCLUSION: This study provides evidence for significant compositional and functional alterations of the gut microbiome in patients with LAA stroke. Gut microbiota might serve as a potential biomarker for patients with LAA stroke.
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Microbioma Gastrointestinal , Accidente Cerebrovascular , Estudios de Casos y Controles , Microbioma Gastrointestinal/fisiología , Humanos , Accidente Cerebrovascular/microbiologíaRESUMEN
BACKGROUND: The gut microbiota (GM)-bone axis has emerged as a crucial mediator of bone homeostasis. Estrogen deficiency-induced bone loss is closely associated with an altered GM. However, the underlying mechanisms remain unclear. OBJECTIVES: We sought to explore the putative effects of GM on estrogen deficiency-induced bone loss and determine a potential mechanism. METHODS: Fecal samples collected from postmenopausal women with osteoporosis (PMO) and with normal bone mass (PMN) were examined by 16S ribosomal RNA (rRNA) gene sequencing and analysis. Prevotella histicola, a typical species of Prevotella, was orally given to female C57BL6/J mice after ovariectomy [ovariectomized (OVX)]. The primary outcomes were changes in bone microstructures as measured by micro-computed tomography scanning and bone histomorphometry analysis. Secondary outcomes included changes in osteoclast activity, the expression of osteoclastogenic cytokines, and gut permeability, which were measured by ELISA, qRT-PCR, western blot, and immunofluorescence. RESULTS: As demonstrated through 16S rRNA gene sequencing and analysis, the GM in the PMO group featured a significantly decreased proportion of the genus Prevotella in comparison with that in the PMN group (â¼60%, P < 0.05). In animal experiments, P. histicola-treated OVX mice maintained a relatively higher bone volume than OVX controls. Mechanistically, the protective effects of P. histicola on bone mass were found to be associated with its modulation of gut permeability as well as its inhibitory effects on osteoclast activity which function by attenuating osteoclastogenic cytokine expression. CONCLUSIONS: The GM diversity and composition between the PMN and PMO groups were significantly different. In particular, the proportion of the genus Prevotella was notably higher in the PMN group, demonstrating its potential bone-protective effects on osteoporosis. Further animal study using osteoporotic mice showed P. histicola could prevent estrogen deficiency-induced bone loss through the GM-bone axis. Thus, P. histicola may serve as a therapeutic agent or target for osteoporosis treatment.
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Estrógenos/deficiencia , Microbioma Gastrointestinal , Osteoporosis/etiología , Ovariectomía/efectos adversos , Posmenopausia , Prevotella , Animales , Citocinas/genética , Citocinas/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Osteoprotegerina/genética , Osteoprotegerina/metabolismo , Ligando RANK/genética , Ligando RANK/metabolismo , ARN Ribosómico 16S/genética , Distribución Aleatoria , Receptor Activador del Factor Nuclear kappa-B/genética , Receptor Activador del Factor Nuclear kappa-B/metabolismoRESUMEN
BACKGROUND: Estrogen deficiency-induced depression is closely associated with an imbalance in intestinal microbiota and inflammation. Prevotella histicola (P. histicola), an emerging probiotic, apparently improves inflammatory responses. This study aims to verify the antidepressant-like effects of P. histicola and clarify its potential mechanisms. METHODS: Mice were treated with P. histicola and cohousing after ovariectomy (OVX). The changes in depression-like behaviors among mice were examined by behavioral tasks, and alterations in the microbiota were detected through 16S rRNA sequencing. Changes in neuronal injury, protein synthesis, inflammatory factors, intestinal permeability, and nerve proliferation were observed by H&E, Nissl staining, qRT-PCR, western blotting, and immunofluorescence. RESULTS: P. histicola significantly reduces depression-like behaviors and neuronal damage induced by estrogen deficiency. Additionally, P. histicola significantly increases the abundance of intestinal flora, especially Lactobacillus and Akkermansia. Meanwhile, the cohoused mice also had a better emotional state and neutral structure compared with OVX mice. P. histicola was also found to upregulate tight junction proteins ZO-1, occludin, claudin-1, and MUC2 in the ileum and colon and reduce the levels of inflammatory factors VCAM, MCP-1, IL-6, IL-8, and TNF-α, mainly in the ileum, colon, and decrease the expression of COX-2, TLR4, Myd88, JNK, MCP-1, IL-6, IL-8, and TNF-α in the hippocampus. Moreover, significant downregulation of apoptosis (caspase-3 and caspase-8) and upregulation of neurotrophic factors (BDNF and Ki-67) were observed after P. histicola treatment. CONCLUSION: Our data show that P. histicola significantly mitigates depression of OVX mice through improvement in intestinal microbiota to repair intestinal leakage and inhibit central inflammation to promote the expression of BDNF for hippocampal neurogenesis. P. histicola may be therapeutically beneficial for PMD.
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OBJECTIVE: To develop anthropometric prediction equations for estimating appendicular skeletal muscle (ASM) in Chinese knee osteoarthritis patients. METHODS: Subjects were divided into the model development group (MD group: 104 cases, 47 men and 57 women) and cross-validation group (CV group: 69 cases, 38 men and 31 women). Stepwise multiple linear regression analyses were undertaken in the MD group to identify the best equations. Agreement between the estimated ASM and ASM measured by dual-energy X-ray absorptiometry (DXA) was tested in the CV group. RESULTS: Two models were developed in the MD group. Validation in the CV group showed that our models (R2 = 0.83 and R2 = 0.90) had a high coefficient of determination. The mean bias of ASM estimated by the two models from the ASM measured by DXA in the CV group showed no significant difference (P > 0.05). CONCLUSION: These models could be useful for older Chinese patients with knee osteoarthritis to estimate ASM.
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Absorciometría de Fotón/métodos , Composición Corporal/fisiología , Músculo Esquelético/fisiopatología , Osteoartritis de la Rodilla/diagnóstico , Sarcopenia/etnología , Factores de Edad , Anciano , Anciano de 80 o más Años , Antropometría , China , Estudios de Cohortes , Femenino , Evaluación Geriátrica , Humanos , Modelos Lineales , Masculino , Análisis Multivariante , Osteoartritis de la Rodilla/epidemiología , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Medición de Riesgo , Sarcopenia/diagnóstico por imagen , Sarcopenia/fisiopatología , Factores SexualesRESUMEN
Tendon to bone (enthesis) rupture, which may cause disability and persistent pain, shows high rate of re-rupture after surgical repair. Tendon or enthesis scaffolds have been widely studied, but few of these materials can recapitulate the tissue continuity. Thus, this study is conducted to prepare a triphasic decellularized bone-fibrocartilage-tendon (D-BFT) composite scaffold. The D-BFT scaffold is developed using a combination of physical, chemical, and enzymatic treatments using liquid nitrogen, Triton-X 100, sodium-dodecyl sulfate, and DNase I, which effectively removes the cell components while preserving the biological composite and microstructure. Moreover, the mechanical properties of D-BFT are highly preserved and similar to those of the human Achilles tendon. Additionally, in vitro, mesenchymal stem cells (MSCs) adhered, proliferated, and infiltrated into the D-BFT scaffold, and MSC differentiation is confirmed by up-regulation of osteogenic-related and tenogenic-related genes. The repair outcomes are explored by applying the D-BFT scaffold in the model of femur-tibia defects in vivo, which shows good repair results. Thus, the D-BFT scaffold developed in this study is a promising graft for enthesis regeneration.
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Tendón Calcáneo/fisiología , Huesos/fisiología , Matriz Extracelular/química , Fibrocartílago/fisiología , Regeneración , Andamios del Tejido/química , Animales , Adhesión Celular , Diferenciación Celular , Proliferación Celular , Colágeno/química , Humanos , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos C57BL , Nitrógeno/química , Osteogénesis , Medicina Regenerativa/instrumentación , Medicina Regenerativa/métodos , Estrés Mecánico , Ingeniería de Tejidos/métodos , Microtomografía por Rayos XRESUMEN
Critical-sized bone defects are incapable of self-healing and are commonly seen in clinical practice. The authors explore a new treatment for this, decellularized periosteum is applied to chitosan globules (chitosan-DP globules) as a hybrid material. The efficacy of chitosan-DP globules on rabbit femoral condyle bone defects is assessed with biocompatibility, biomechanics, and osteogenic efficiency measurements, and compared with the results of chitosan globules and empty control. No difference in cytotoxicity is observed among chitosan-DP globules, chitosan globules, and the empty control. Chitosan-DP globules possesse a better surface for cell adhesion than did chitosan globules. Chitosan-DP globules demonstrate superior efficiency for osteogenesis in the defect area compared to chitosan globules as per microcomputed tomography examination and push-out testing, with relatively minor histological differences. Both chitosan globule groups show more satisfactory results than those for the empty control. The results implicate chitosan-DP globules as a promising solution for bone defects.
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Materiales Biocompatibles/farmacología , Regeneración Ósea/efectos de los fármacos , Quitosano/farmacología , Fémur/patología , Periostio , Animales , Materiales Biocompatibles/química , Fenómenos Biomecánicos , Regeneración Ósea/fisiología , Adhesión Celular/efectos de los fármacos , Células Cultivadas , Quitosano/farmacocinética , Fémur/lesiones , Células Madre Mesenquimatosas/efectos de los fármacos , Peso Molecular , Conejos , Propiedades de Superficie , Microtomografía por Rayos XRESUMEN
To explore the effects of captopril on calpainmediated apoptosis of myocardial cells and cardiac function in diabetic rats, 30 adult male SpragueDawley rats were randomly divided into three groups: Negative control (NC group), untreated diabetic rats (DM group) and diabetic rats treated with captopril (Cap group). Diabetes was induced by streptozotocin injection. Captopril was intragastrically administered at a daily dose of 50 mg/kg for 12 weeks; the NC and DM groups received an equivalent volume of saline. After 12 weeks of treatment, left ventricular systolic pressure (LVSP), left ventricular enddiastolic pressure (LVDEP), maximal rate of left ventricular pressure increase (+dp/dtmax), maximal rate of left ventricular pressure decrease (dp/dtmax) and left ventricular mass index (LVMI) were measured. The levels of calpain1, calpain2, Bcell lymphoma (Bcl)2, Bcl2 associated protein X (Bax) and total caspase3 were detected in cardiac tissue by western blot analysis. The apoptotic index (AI) was assessed with a terminal deoxynucleotidyl transferasemediated dUTP nickend labeling assay. The ultrastructure of cardiac tissue was determined by transmission electron microscopy. Compared with the NC group, LVDEP and LVMI were increased, whereas LVSP, +dp/dtmax and dp/dtmax were decreased in the DM group. In the Cap group, LVDEP and LVMI were decreased, whereas LVSP, +dp/dtmax and dp/dtmax were increased compared with the DM group. Bcl2 protein expression was decreased, whereas the levels of calpain1, calpain2, Bax and total caspase3 protein were increased in the DM group, compared with the NC group. Cap treatment increased Bcl2 protein expression and decreased calpain1, calpain2, Bax and total caspase3 protein expression compared with the DM group. Additionally, the AI was increased in the DM group compared with the NC group, and decreased in the Cap group compared with the DM group. Furthermore, ultrastructural examination demonstrated that myocardial cell injury was reduced in the Cap group compared with the DM group. Therefore, captopril improved myocardial structure and ventricular function, by inhibiting calpain1 and calpain2 activation, increasing Bcl2 expression, reducing Bax expression and subsequently inhibiting caspase3dependent apoptosis.
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Captopril/administración & dosificación , Diabetes Mellitus Experimental/tratamiento farmacológico , Corazón/efectos de los fármacos , Disfunción Ventricular Izquierda/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Calpaína/efectos adversos , Calpaína/genética , Caspasa 3/genética , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Corazón/fisiopatología , Humanos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Ratas , Disfunción Ventricular Izquierda/genética , Disfunción Ventricular Izquierda/patología , Proteína X Asociada a bcl-2/genéticaRESUMEN
The occurrence and the subsequent development of pulmonary arterial hypertension (PAH) involve complicated mechanisms. Of these, the proliferation of pulmonary artery smooth muscle cells (PASMCs) has been indicated to be closely associated with its progression. Therefore, therapeutic methods targeting PASMCs to inhibit proliferation is an effective method for alleviating PAH. The present study was designed to determine the role of the adenosine A(2A) receptor (A2A receptor) in hypoxiainduced rat PASMC (RPASMC) proliferation. Primary RPASMCs were isolated from the pulmonary artery of adult male SD rats, cultured and used for the following experiments. The mRNA level and protein expression of CXCR4 were measured by reverse transcriptionquantitative polymerase chain reaction and western blot analysis, respectively. The cell proliferation of RPASMCs was measured using a cell proliferation assay kit. In the present study, it was demonstrated that the proliferation of RPASMCs was partially mediated by activation of the stromal cellderived factor 1 (SDF1)CXC chemokine receptor 4 (CXCR4) axis under hypoxic conditions. In addition, SDF1α alone upregulated the mRNA and protein expression levels of CXCR4, and stimulated the proliferation of RPASMCs. The protein expression of CXCR4 and the cell proliferation were markedly inhibited by application of A2A receptor agonist CGS21680 or cyclic adenosine monophosphate (cAMP) under hypoxic conditions or treatment with SDF1α and was reversed by the A2A receptor antagonist SCH58261 or 8bromoadenosine3',5'cyclic monophosphorothioate. These results demonstrated that the inhibition of SDF1CXC4 signaling by the activation of A2A receptor and subsequent increase in the level of cAMP may be a potential method to ameliorate PAH.
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Quimiocina CXCL12/metabolismo , AMP Cíclico/metabolismo , Miocitos del Músculo Liso/metabolismo , Arteria Pulmonar/patología , Receptor de Adenosina A2A/metabolismo , Receptores CXCR4/metabolismo , Transducción de Señal , Animales , Hipoxia de la Célula , Proliferación Celular , Masculino , Ratas Sprague-Dawley , Regulación hacia ArribaRESUMEN
AIMS/INTRODUCTION: In previous studies, hydrogen gas (H2) administration has clearly shown effectiveness in inhibiting diabetes. Here, we evaluated whether subcutaneous injection of H2 shows enhanced efficacy against type 2 diabetes mellitus induced in mice by a high-fat diet and low-dose streptozotocin treatment. MATERIAL AND METHODS: H2 was injected subcutaneously at a dose of 1 mL/mouse/week for 4 weeks. Type 2 diabetes mellitus-associated parameters were then evaluated to determine the effectiveness of subcutaneous H2 administration. RESULTS: The bodyweight of H2 -treated mice did not change over the course of the experiment. Compared with the untreated control animals, glucose, insulin, low-density lipoprotein and triglyceride levels in the serum were significantly lower in treated mice, whereas high-density lipoprotein cholesterol in the serum was significantly higher. Glucose tolerance and insulin sensitivity were both improved in H2 -treated mice. Diabetic nephropathy analysis showed significant reductions in urine volume, urinary total protein and ß2-microglobulin, kidney/bodyweight ratio, and kidney fibrosis associated with subcutaneous injection of H2 . CONCLUSIONS: Subcutaneous injection of H2 significantly improves type 2 diabetes mellitus and diabetic nephropathy-related outcomes in a mouse model, supporting further consideration of subcutaneous injection as a novel and effective route of clinical H2 administration.
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Diabetes Mellitus Tipo 2/terapia , Hidrógeno/administración & dosificación , Animales , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/metabolismo , Nefropatías Diabéticas/patología , Nefropatías Diabéticas/terapia , Dieta Alta en Grasa , Inyecciones Subcutáneas , Masculino , Ratones Endogámicos C57BL , Estrés Oxidativo , EstreptozocinaRESUMEN
Ischemia/reperfusion (I/R) injury plays an essential role in renal transplantation, and represents a crucial risk factor for allograft dysfunction and acute renal failure. Modulation of oxidative stress is an effective therapeutic strategy for I/R injury. Perillyl alcohol (POH), a dietary monoterpene with antioxidant activity is found in a variety of plants. The study was carried out to investigate whether treatment of POH could reduce hypoxia/reoxygenation (H/R)-induced injury. H/R induced significant injury in HK-2 cells. H/R caused an increase in ROS level, apoptosis and ER stress. Meanwhile H/R also inhibited the cell viability and PI3K/Akt/eNOS signaling pathway. Pretreatment with POH prior to H/R improved cell viability, reduce ROS level, ER stress and apoptosis. Moreover, POH could also activate the PI3K/Akt/eNOS pathway. Therefore, POH may possess protective effects in H/R-induced cellular damage.
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Estrés del Retículo Endoplásmico/efectos de los fármacos , Células Epiteliales/patología , Túbulos Renales/patología , Monoterpenos/farmacología , Oxígeno/toxicidad , Sustancias Protectoras/farmacología , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacos , Hipoxia de la Célula/efectos de los fármacos , Línea Celular , Citoprotección/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Óxido Nítrico Sintasa de Tipo III/metabolismo , Estrés Oxidativo/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismoRESUMEN
Osteonecrosis of the femoral head(ONFH), a refractory disease characterized by death of the osteocytes and the bone marrow due to inadequate blood supply caused by various mechanisms, usually leads to the collapse of the femoral head and malfunction of the hip joint. The crux is to diagnose ONFH early in the precollapse stage and prevent subsequent progression of collapsing through early interventions, thus delaying or avoiding the replacement of the hip joint. A number of joint salvaging operation treatments for early stage ONFH are available. However, there has been no consensus with regard to the ideal treatment. The main trend now is to unite core decompression with bone-grafting, tantalum rod, bone marrow mesenchymal stem cell (BMSC) and other treatments. Also there are ways of osteotomy altering the angle of the femoral neck to relocate necrotic tissue from the weight-bearing segment. The implanting of tantalum rod remains controversial and the advent of bone marrow mesenchymal stem cell (BMSC) holds huge potential.
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Necrosis de la Cabeza Femoral/diagnóstico , Necrosis de la Cabeza Femoral/prevención & control , Terapia Recuperativa/métodos , Trasplante Óseo , Descompresión Quirúrgica , Progresión de la Enfermedad , Cuello Femoral , Humanos , Trasplante de Células Madre Mesenquimatosas , TantalioRESUMEN
Hypericin (Hyp) is traditionally used as an antidepressant and antiviral agent. It selectively accumulates in spheroids and is also used as a photosensitizer in the photodynamic therapy of cancer. The present study aimed to investigate the cytotoxic effect of Hypmediated photodynamic therapy (HypPDT) on cell growth and apoptosis of K562 leukemia cells, and to examine the underlying mechanisms. HypPDT was performed with different light intensities (0.1, 0.3 and 0.5 mW/cm2), different concentrations of Hyp (0, 0.2, 0.4 and 0.8 µg/ml) and different durations of irradiation (0, 2, 4 and 8 min) in order to select the optimal conditions for subsequent experiments. A concentration of 0.4 µg/ml Hyp with a 5 h druglight interval and 4 min irradiation at 0.3 mW/cm2 light intensity was selected as the optimal conditions. The effects of HypPDT on apoptosis were determined by detecting morphological changes under microscopy and by performing western blot analysis. The results revealed that HypPDT suppressed cell viability in a light intensity, dose and irradiation durationdependent manner. The expression levels of cleaved caspase9, cleaved caspase3 and phosphorylatedCJun N terminal kinase (JNK)l were significantly upregulated following HypPDT. These results indicated that HypPDT decreased cell viability and induced mitochondriacaspasedependent apoptosis in the K562 cells through regulation of the JNK pathway. These findings suggest that Hyp-PDT may be developed as an effective treatment for leukemia.
Asunto(s)
Apoptosis/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica , Perileno/análogos & derivados , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Antracenos , Caspasa 3/genética , Caspasa 3/metabolismo , Caspasa 9/genética , Caspasa 9/metabolismo , Supervivencia Celular , Humanos , Células K562 , Luz , MAP Quinasa Quinasa 4/genética , MAP Quinasa Quinasa 4/metabolismo , Perileno/farmacología , Transducción de SeñalRESUMEN
Bone grafting or bone substitute is typically used to bridge a bone defect that has been caused by trauma, tumor resection, pathological degeneration, or congenital deformations. However, bone graft healing and remodeling is always a major concern of orthopedic surgeons. Because the periosteum has a remarkable regenerative capacity and is widely recognized to be essential for the initiation of bone graft healing and remodeling, the present study aimed to produce a rabbit decellularized periosteum (D-periosteum) to be used as a biologic scaffold for future bone tissue engineering. We obtained the D-periosteum by employing a combination of commonly used decellularization processes, which include physical methods as well as chemical and enzymatic solutions. The cellular components were effectively removed, and this removal was demonstrated using current decellularization criteria (H&E staining, DAPI staining, DNA quantification and agarose gel electrophoresis); however, there were no significant alterations of the native extracellular matrix (ECM) properties (collagen, glycosaminoglycan (GAG), microarchitecture and mechanical properties). Periosteum-derived cells (PDCs) could adhere, proliferate and infiltrate into the D-periosteum in vitro. The allogenic D-periosteum was implanted subcutaneously into the backs of rabbits over 28 days to study the biocompatibility in vivo. The D-periosteum did not elicit a severe immunogenic response. In summary, a biologic scaffold composed of ECM from periosteum has been successfully developed. The D-periosteum maintains biocompatibility in vitro and in vivo and, therefore, can provide a naturally compatible scaffold for use in future bone tissue engineering.