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1.
Life Sci ; 244: 117342, 2020 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-31978450

RESUMEN

AIMS: Microvascular endothelial cell dysfunction is a leading cause of radiation-induced heart disease (RIHD). BRCA1 plays an important role in DNA damage repair. The study aims to explore the effect of BRCA1 in endothelial cells involved in RIHD. MATERIALS AND METHODS: BRCA1 and p21 expression were detected in human umbilical vein endothelial cells (HUVECs) and in mouse heart tissue after irradiation exposure. The effects of BRCA1 on cell proliferation, cell cycle and radiosensitivity were determined in HUVECs with overexpression and knockdown of BRCA1. A mouse model of RIHD was established. Heart damage was detected in C57BL/6J mice and endothelial cell specific knockout BRCA1 mice (EC-BRCA1-/-). KEY FINDINGS: BRCA1 and p21 expression was significantly increased both in vitro and vivo response to irradiation. BRCA1 overexpression in endothelial cells enhanced cell growth and G1/S phase arrest, and the opposite results were observed in BRCA1 knockdown endothelial cells. BRCA1 downregulated endothelial cell cycle-related genes cyclin A, cyclin D1, cyclin E and p-Rb through increasing p21 expression, and HUVECs with BRCA1 gene knockdown were more sensitive to radiation. In vivo, a decrease in cardiac microvascular density, as well as cardiomyocyte hypoxia and apoptosis were observed in a time-dependent manner. EC-BRCA1-/- mice were more prone to severe RIHD than EC-BRCA1+/- mice after 16Gy radiation exposure due to endothelial dysfunction caused by loss of BRCA1, and p21 was declined in EC-BRCA1-/- mice heart. SIGNIFICANCE: These findings indicate that BRCA1 plays a protective role in RIHD by regulating endothelial cell cycle arrest mediated by p21 signal.


Asunto(s)
Proteína BRCA1/metabolismo , Puntos de Control del Ciclo Celular , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Células Endoteliales de la Vena Umbilical Humana/citología , Neovascularización Patológica/prevención & control , Sustancias Protectoras/administración & dosificación , Tolerancia a Radiación , Animales , Proteína BRCA1/genética , Proteína BRCA1/fisiología , Proliferación Celular , Células Cultivadas , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/efectos de la radiación , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neovascularización Patológica/etiología , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Radiación Ionizante
2.
Toxicol Lett ; 318: 12-21, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31622651

RESUMEN

Maternal smoking during pregnancy and lactation is associated with increased fat mass in the offspring, but the mechanism by which this occurs is not fully understood. Our study focused on the relationships among maternal nicotine exposure, adipose angiogenesis and adipose tissue function in female offspring. Pregnant rats were randomly assigned to nicotine or control groups. Microvascular density, lipid metabolism and α7nAChR-Egr1-FGF2 signaling pathway genes/proteins were tested in 4-, 12- and 26-week female offspring. In vitro, nicotine concentration- and time-response experiments were conducted in 3T3-L1. Lipid metabolism and α7nAChR-Egr1-FGF2 signaling pathway genes/proteins were tested. The conditioned media of differentiated 3T3-L1 treated with nicotine were used to observe tube formation in human umbilical vein endothelial cells (HUVECs). Nicotine-exposed females presented higher adipose microvascular density. The gene expression of α7nAChR, Egr1 and FGF2 was significantly increased in gonadal white adipose tissue (gWAT) and inguinal subcutaneous WAT (igSWAT) of nicotine-exposed females at 4 weeks of age. The protein expression of α7nAChR, Egr1 and FGF2 was increased in gWAT and igSWAT of nicotine-exposed females at 4 weeks of age, and increased in gWAT at 26 weeks. In vitro, nicotine increased the expression of lipid metabolism and α7nAChR-Egr1-FGF2 signaling pathway genes/proteins in a concentration- and time-dependent manner. In the tube formation experiment, adipocytes affected by nicotine promoted HUVEC angiogenesis. Therefore, maternal nicotine exposure promoted the early angiogenesis of adipose tissue via the α7nAChR-Egr1-FGF2 signaling pathway, and this angiogenesis mechanism was associated with increased adipogenesis in adipose tissue of female offspring.


Asunto(s)
Adipocitos/efectos de los fármacos , Tejido Adiposo Blanco/irrigación sanguínea , Neovascularización Fisiológica/efectos de los fármacos , Nicotina/toxicidad , Agonistas Nicotínicos/toxicidad , Efectos Tardíos de la Exposición Prenatal , Células 3T3-L1 , Adipocitos/metabolismo , Animales , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Proteína 1 de la Respuesta de Crecimiento Precoz/metabolismo , Femenino , Factor 2 de Crecimiento de Fibroblastos/genética , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , Exposición Materna , Ratones , Embarazo , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Receptor Nicotínico de Acetilcolina alfa 7/genética , Receptor Nicotínico de Acetilcolina alfa 7/metabolismo
3.
Int J Nanomedicine ; 14: 8973-8987, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31819413

RESUMEN

Background: Elevated levels of low density lipoprotein (LDL), "bad cholesterol", is not an accurate indicator of coronary disease. About 75% of patients with heart attacks have cholesterol levels that do not indicate a high risk for a cardiovascular event. LDL is comprised of three subclasses, with particles of different size and density. We used nanomedical systems to elucidate the noxious effects of LDL subclasses on endothelium. Experimental: Nanosensors were employed to measure the concentrations of nitric oxide (NO) and peroxynitrite (ONOO-) stimulated by LDL subclasses in HUVECs. N-LDL and ox-LDL (subclass A: 1.016-1.019 g/mL, subclass I: 1.024-1.029 g/mL, and subclass B: 1.034-1.053 g/mL) stimulated NO and ONOO- release. The concentrations ratio of (NO)/(ONOO-) was used to evaluate the noxious effects of the subclasses on endothelium. Results: In HUVECs, the (NO)/(ONOO-) ratio for normal endothelium is about 5, but shifts to 2.7±0.4, 0.5±0.1, and 0.9±0.1 for subclasses A, B, and I, respectively. Ratios below 1.0 indicate an imbalance between NO and ONOO-, affecting endothelial function. LDL of 50% B and 50% I produced the most severe imbalance (0.45±0.04), whereas LDL of 60% A, 20% B, and 20% I had the most favorable balance of 5.66±0.69. Subclass B significantly elevated the adhesion of molecules and monocytes. The noxious effect was significantly higher for ox-LDL than n-LDL. Conclusion: Subclass B of "bad cholesterol" is the most damaging to endothelial function and can contribute to the development of atherosclerosis. Contrary to the current national guidelines, this study suggests that it's not the total LDL, rather it is the concentration of subclass B in relation to subclasses A and/or I, that should be used for diagnosis of atherosclerosis and the risk of heart attack. By utilizing specific pharmacological therapy to address the concentration of subclass B, there is a potential to significantly reduce the risk of heart attack and atherosclerosis.


Asunto(s)
Enfermedades Cardiovasculares/metabolismo , Colesterol/metabolismo , Lipoproteínas LDL/metabolismo , Ácido Peroxinitroso/metabolismo , Enfermedades Cardiovasculares/patología , Adhesión Celular , Femenino , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Monocitos/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Células THP-1
4.
Int J Nanomedicine ; 14: 9285-9294, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31819430

RESUMEN

Purpose: Physicochemical properties play a crucial role in determining the toxicity of multi-walled carbon nanotubes (MWCNTs). Recently we found that MWCNTs with longer length and smaller diameters could induce toxicity to human umbilical vein endothelial cells (HUVECs) through the activation of endoplasmic reticulum (ER) stress. In this study, we further investigated the possible contribution of hydroxylation and carboxylation to the cytotoxicity of MWCNTs. Methods: The HUVECs were exposed to pristine (code XFM19), hydroxylated (code XFM20; content of hydroxyl groups 1.76 wt%) and carboxylated (code XFM21; content of carboxyl groups 1.23 wt%) MWCNTs, respectively. Then, the internalization, cytotoxicity, oxidative stress and activation of apoptosis-ER stress pathway were measured. Results: In consequence, all types of MWCNTs could be internalized into the HUVECs, and the cellular viability was significantly reduced to a similar level. Moreover, the MWCNTs increased intracellular reactive oxygen species (ROS) and decreased glutathione (GSH) to similar levels, indicating their capacity of inducing oxidative stress. The Western blot results showed that all types of MWCNTs reduced BCL-2 and increased caspase-3, caspase-8, cleaved caspase-3 and cleaved caspase-8. The expression of ER stress gene DNA damage-inducible transcript 3 (DDIT3) and protein level of chop were only significantly induced by XFM20 and XFM21, whereas protein level of p-chop was promoted by XFM19 and XFM21. In addition, the pro-survival gene XBP-1s was significantly down-regulated by all types of MWCNTs. Conclusion: These results suggested that MWCNTs could induce cytotoxicity to HUVECs via the induction of oxidative stress and apoptosis-ER stress, whereas a low degree of hydroxylation or carboxylation did not affect the toxicity of MWCNTs to HUVECs.


Asunto(s)
Apoptosis , Estrés del Retículo Endoplásmico , Nanotubos de Carbono/química , Supervivencia Celular , Endocitosis , Estrés del Retículo Endoplásmico/genética , Glutatión/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Hidrodinámica , Nanotubos de Carbono/ultraestructura , Estrés Oxidativo , Tamaño de la Partícula , Especies Reactivas de Oxígeno/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , Electricidad Estática
5.
Adv Clin Exp Med ; 28(11): 1441-1450, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31778596

RESUMEN

BACKGROUND: Human cytomegalovirus (HCMV) infection is one of the risk factors of cardiovascular disease; the most important pathological change is the change of vascular endothelial cell (VEC) function, but its mechanism is still unclear. Transforming growth factor ß1 (TGF-ß1) is an important cytokine associated with fibrosis; it can induce the occurrence of endothelial mesenchymal transition (EndMT) in VECs, which means endothelial cells acquire the characteristics and phenotypes of mesenchymal cells and secrete molecules associated with the deposition and remodeling of the extracellular matrix. Many in vivo and in vitro studies have shown that HCMV infection promotes the secretion and activation of TGF-ß1. OBJECTIVES: This study aims to observe the changes of endothelial cells after HCMV infection and EndMT occurrence induced by TGF-ß1 and to explore the possible mechanism of HCMV infection in the pathogenesis of cardiovascular disease. MATERIAL AND METHODS: Immunofluorescence staining, reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and immunoprecipitation methods were used in this study to analyze the changes in morphology and gene expression. RESULTS: We found that EndMT-related morphological and gene expression changes occurred in human umbilical vein endothelial cells (HUVECs) infected and uninfected with HCMV after treatment with TGF-ß1. Human umbilical vein endothelial cells infected with HCMV, which are treated with TGF-ß1, can activate the extracellular potential TGF-ß1 by activating matrix metalloproteinase 2 (MMP-2). CONCLUSIONS: Our findings provide a molecular basis for the association between HCMV infection, TGF-ß1 and cardiovascular disease.


Asunto(s)
Infecciones por Citomegalovirus/tratamiento farmacológico , Citomegalovirus , Transición Epitelial-Mesenquimal , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Metaloproteinasa 2 de la Matriz , Factor de Crecimiento Transformador beta1/metabolismo , Factor de Crecimiento Transformador beta1/farmacología , Venas Umbilicales/metabolismo , Venas Umbilicales/virología , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Infecciones por Citomegalovirus/diagnóstico , Transición Epitelial-Mesenquimal/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Transducción de Señal
6.
DNA Cell Biol ; 38(12): 1557-1563, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31580158

RESUMEN

Activation of TLR4-MyD88-NF-κB signaling by lipopolysaccharide (LPS) evokes a proinflammatory immune response, and plays a pivotal role in initiation and progression of atherosclerosis (AS). ATP (adenosine 5'-trisphosphate), a powerful extracellular signal transduction molecule, functions to regulate immune inflammatory responses depending on the type of P2 receptors and cell lines. In this study, we first performed RT-PCR to detect the mRNA expression of monocyte chemoattractant protein-1 (MCP-1), IL-8, and IL-1ß induced by different concentrations of LPS in human umbilical vein endothelial cells (HUVECs). Protein level of TLR4 signaling including TLR4, myeloid differentiation factor (MyD88), and CD14 induced by LPS (1 µg/mL) at different times (0, 10, 30, 60, 120 min) was analyzed by Western blot. Then, RT-PCR was performed to detect the effect of different concentrations of ATP on mRNA expression of IL-1ß and MCP-1 induced by LPS (1 µg/mL) and the TLR4 signaling pathway. Western blot was performed to detect the effect of low concentrations of ATP on phosphorylation of p65 induced by 1 µg/mL LPS. Finally, we used P2Y receptor blocker Suramin to verify whether the role of ATP on LPS-induced inflammatory cytokine expression was through P2Y receptors. The results showed that LPS upregulated the expression of MCP-1, IL-8, and IL-1ß in a dose-dependent manner accompanied by the activation of TLR4-MyD88 signaling in HUVECs. Only low concentration ATP (1, 10 µM) inhibited LPS-induced mRNA expression of IL-1ß and MCP-1. ATP at low concentrations also downregulated the mRNA expression of TLR4, CD14, and MyD88 and inhibited LPS-induced phosphorylation of p65. Furthermore, Suramin, a nonspecific P2Y receptor antagonist, did not attenuate the inhibition of ATP on LPS-induced IL-1ß and MCP-1 expression. Taking this together, low concentration ATP inhibited LPS-induced inflammation in HUVECs by negatively regulating TLR4-MyD88 signaling, and P2Y receptors were not involved in this process, which might provide new ideas for prevention and treatment of inflammatory diseases such as AS.


Asunto(s)
Adenosina Trifosfato/farmacología , Antiinflamatorios/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Inflamación/tratamiento farmacológico , Lipopolisacáridos/farmacología , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Receptor Toll-Like 4/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Inflamación/inducido químicamente , Inflamación/metabolismo , Inflamación/patología , Factor 88 de Diferenciación Mieloide/genética , FN-kappa B/genética , Receptor Toll-Like 4/genética
7.
Int J Mol Sci ; 20(19)2019 Oct 05.
Artículo en Inglés | MEDLINE | ID: mdl-31590384

RESUMEN

Loss-of-function mutations of the gene encoding Krev interaction trapped protein 1 (KRIT1) are associated with the pathogenesis of Cerebral Cavernous Malformation (CCM), a major cerebrovascular disease characterized by abnormally enlarged and leaky capillaries and affecting 0.5% of the human population. However, growing evidence demonstrates that KRIT1 is implicated in the modulation of major redox-sensitive signaling pathways and mechanisms involved in adaptive responses to oxidative stress and inflammation, suggesting that its loss-of-function mutations may have pathological effects not limited to CCM disease. The aim of this study was to address whether KRIT1 loss-of-function predisposes to the development of pathological conditions associated with enhanced endothelial cell susceptibility to oxidative stress and inflammation, such as arterial endothelial dysfunction (ED) and atherosclerosis. Silencing of KRIT1 in human aortic endothelial cells (HAECs), coronary artery endothelial cells (HCAECs), and umbilical vein endothelial cells (HUVECs) resulted in increased expression of endothelial proinflammatory adhesion molecules vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) and in enhanced susceptibility to tumor necrosis factor alpha (TNF-α)-induced apoptosis. These effects were associated with a downregulation of Notch1 activation that could be rescued by antioxidant treatment, suggesting that they are consequent to altered intracellular redox homeostasis induced by KRIT1 loss-of-function. Furthermore, analysis of the aorta of heterozygous KRIT1+/- mice fed a high-fructose diet to induce systemic oxidative stress and inflammation demonstrated a 1.6-fold increased expression of VCAM-1 and an approximately 2-fold enhanced fat accumulation (7.5% vs 3.6%) in atherosclerosis-prone regions, including the aortic arch and aortic root, as compared to corresponding wild-type littermates. In conclusion, we found that KRIT1 deficiency promotes ED, suggesting that, besides CCM, KRIT1 may be implicated in genetic susceptibility to the development of atherosclerotic lesions.


Asunto(s)
Aorta/metabolismo , Aterosclerosis/genética , Endotelio Vascular/metabolismo , Proteína KRIT1/genética , Mutación con Pérdida de Función , Animales , Aorta/patología , Apoptosis , Aterosclerosis/metabolismo , Endotelio Vascular/patología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Proteína KRIT1/deficiencia , Proteína KRIT1/metabolismo , Metabolismo de los Lípidos , Ratones , Ratones Endogámicos C57BL , Estrés Oxidativo , Receptor Notch1/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Molécula 1 de Adhesión Celular Vascular/metabolismo
8.
Int J Nanomedicine ; 14: 7399-7417, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31571858

RESUMEN

Purpose: We studied the effects of silver nanoparticles (AgNPs) on human blood platelet function. We hypothesized that AgNPs, a known antimicrobial agent, can be used as blood-compatible, "ideal material'' in medical devices or as a drug delivery system. Therefore, the aim of the current study was to investigate if functionalized AgNPs affect platelet function and platelets as well as endothelial cell viability in vitro. Methods: AgNPs, functionalized with reduced glutathione (GSH), polyethylene glycol (PEG) and lipoic acid (LA) were synthesized. Quartz crystal microbalance with dissipation was used to measure the effect of AgNPs on platelet aggregation. Platelet aggregation was measured by changes in frequency and dissipation, and the presence of platelets on the sensor surface was confirmed and imaged by phase contrast microscopy. Flow cytometry was used to detect surface abundance of platelet receptors. Lactate dehydrogenase test was used to assess the potential cytotoxicity of AgNPs on human blood platelets, endothelial cells, and fibroblasts. Commercially available ELISA tests were used to measure the levels of thromboxane B2 and metalloproteinases (MMP-1, MMP-2) released by platelets as markers of platelet activation. Results: 2 nm AgNPs-GSH, 3.7 nm AgNPs-PEG both at 50 and 100 µg/mL, and 2.5 nm AgNPs-LA at 100 µg/mL reduced platelet aggregation, inhibited collagen-mediated increase in total P-selectin and GPIIb/IIIa, TXB2 formation, MMP-1, and MMP-2 release. The tested AgNPs concentrations were not cytotoxic as they did not affect, platelet, endothelial cell, or fibroblast viability. Conclusion: All tested functionalized AgNPs inhibited platelet aggregation at nontoxic concentrations. Therefore, functionalized AgNPs can be used as an antiplatelet agent or in design and manufacturing of blood-facing medical devices, such as vascular grafts, stents, heart valves, and catheters.


Asunto(s)
Plaquetas/efectos de los fármacos , Nanopartículas del Metal/química , Agregación Plaquetaria/efectos de los fármacos , Plata/farmacología , Colágeno/metabolismo , Fibroblastos/efectos de los fármacos , Citometría de Flujo , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , L-Lactato Deshidrogenasa/metabolismo , Ligandos , Metaloproteinasas de la Matriz/metabolismo , Nanopartículas del Metal/ultraestructura , Selectina-P/metabolismo , Tamaño de la Partícula , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Polietilenglicoles/química , Tecnicas de Microbalanza del Cristal de Cuarzo , Espectroscopía Infrarroja por Transformada de Fourier , Tromboxano B2/metabolismo
9.
Mol Immunol ; 116: 73-79, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31630078

RESUMEN

Atherosclerosis is a common comorbidity of type II diabetes and a leading cause of death worldwide. The presence of oxidized low-density lipoprotein (ox-LDL) drives atherogenesis by inducing oxidative stress, mitochondrial dysfunction, expression of proinflammatory cytokines and chemokines including interleukin (IL)-1ß, IL-6, and monocyte chemoattractant protein 1 (MCP-1), adhesion molecules including vascular cellular adhesion molecule 1 (VCAM-1) and E-selectin, and downregulating expression of the Krüppel-like factor 2 (KLF2) transcription factor. Importantly, ox-LDL induced the attachment of THP-1 monocytes to endothelial cells. In the present study, we demonstrate for the first time that the specific glucagon-like peptide 1 receptor (GLP-1R) agonist dulaglutide may prevent these atherosclerotic effects of ox-LDL by preventing suppression of KLF2 by p53 protein in human aortic endothelial cells. KLF2 has been shown to play a major role in protecting vascular endothelial cells from damage induced by ox-LDL and oscillatory shear, and therefore, therapies capable of mediating KLF2 signaling may be an attractive treatment option for preventing the development and progression of atherosclerosis.


Asunto(s)
Aterosclerosis/tratamiento farmacológico , Receptor del Péptido 1 Similar al Glucagón/metabolismo , Péptidos Similares al Glucagón/análogos & derivados , Fragmentos Fc de Inmunoglobulinas/farmacología , Lipoproteínas LDL/metabolismo , Monocitos/efectos de los fármacos , Proteínas Recombinantes de Fusión/farmacología , Aterosclerosis/metabolismo , Adhesión Celular , Células Cultivadas , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Células Endoteliales , Regulación de la Expresión Génica/efectos de los fármacos , Péptidos Similares al Glucagón/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Factores de Transcripción de Tipo Kruppel/metabolismo , Monocitos/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Molécula 1 de Adhesión Celular Vascular/metabolismo
10.
J Food Sci ; 84(11): 3284-3295, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31618463

RESUMEN

Pectinase is a well-known enzyme used in the food processing industry to produce fruit juice and concentrate. This study evaluated the anticancer and antiangiogenesis activities of pectinase-treated Prunus mume fruit concentrate (PC) and its phenolic components. PC treatment (250 to 1,000 µg/mL) resulted in decreased proliferation of SW480 human colorectal cancer cells through S-phase cell cycle arrest; however, equivalent concentrations of PC did not show toxicity toward CRL-1539 colon normal cells. Furthermore, PC-induced caspase-dependent apoptosis in SW480 cells, which was characterized by accumulation of apoptotic cell population, cell shrinkage, formation of apoptotic bodies, upregulation of proapoptotic Bax, cleaved PARP, caspase-3, caspase-8, and caspase-9, and downregulation of antiapoptotic Bcl-2. Antiangiogenesis effects of PC were assessed using human umbilical vein endothelial cells (HUVECs). We found that PC did not inhibit HUVECs proliferation at concentrations of 500 to 1,500 µg/mL. In addition, treatment with PC at nontoxic concentrations (500 to 1,000 µg/mL) blocked vascular endothelial growth factor induced cell migration, invasion, capillary-like tube formation, and angiogenesis from rat aortic rings. HPLC-PDA analysis showed that there were at least four different phenolics including 5-HMF, neochlorogenic acid, protocatechuic acid, and syringic acid. Taken together, these results indicated that PC could be used as a good source of phenolic compounds with selective anticancer and antiangiogenesis activities. PRACTICAL APPLICATION: Pectinases are one of the well-known enzyme used in the part of food processing. Treatment of pectinase is a useful strategy to reduce viscosity, turbidity, and pulp particles in the production of fruit juice, extract, and concentrate. In the present study, we found that pectinase-treated P. mume fruit concentrate significantly suppresses colorectal cancer proliferation and angiogenesis of human umbilical vein endothelial cells. The significance of our findings is that pectinase-treated P. mume concentrate may be used as a commercial functional food material to inhibit colorectal cancer and angiogenesis.


Asunto(s)
Inhibidores de la Angiogénesis/química , Inhibidores de la Angiogénesis/farmacología , Neoplasias Colorrectales/fisiopatología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Poligalacturonasa/química , Prunus/química , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Frutas/química , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Hidroxibenzoatos/química , Hidroxibenzoatos/farmacología , Fenoles/química , Fenoles/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo
11.
Oxid Med Cell Longev ; 2019: 1202676, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31531177

RESUMEN

Malaysian Tualang honey (TH) is a known therapeutic honey extracted from the honeycombs of the Tualang tree (Koompassia excelsa) and has been reported for its antioxidant, anti-inflammatory, antiproliferative, and wound healing properties. However, the possible vascular protective effect of TH against oxidative stress remains unclear. In this study, the effects of TH on hydrogen peroxide- (H2O2-) elicited vascular hyperpermeability in human umbilical vein endothelial cells (HUVECs) and Balb/c mice were evaluated. Our data showed that TH concentrations ranging from 0.01% to 1.00% showed no cytotoxic effect to HUVECs. Induction with 0.5 mM H2O2 was found to increase HUVEC permeability, but the effect was significantly reversed attenuated by TH (p < 0.05), of which the permeability with the highest inhibition peaked at 0.1%. In Balb/c mice, TH (0.5 g/kg-1.5 g/kg) significantly (p < 0.05) reduced H2O2 (0.3%)-induced albumin-bound Evans blue leak, in a dose-dependent manner. Immunofluorescence staining confirmed that TH reduced actin stress fiber formation while increasing cortical actin formation and colocalization of caveolin-1 and ß-catenin in HUVECs. Signaling studies showed that HUVECs pretreated with TH significantly (p < 0.05) decreased intracellular calcium release, while sustaining the level of cAMP when challenged with H2O2. These results suggested that TH could inhibit H2O2-induced vascular hyperpermeability in vitro and in vivo by suppression of adherence junction protein redistribution via calcium and cAMP, which could have a therapeutic potential for diseases related to the increase of both oxidant and vascular permeability.


Asunto(s)
Señalización del Calcio/efectos de los fármacos , Permeabilidad Capilar/efectos de los fármacos , Miel , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Peróxido de Hidrógeno/farmacología , AMP Cíclico/metabolismo , Relación Dosis-Respuesta a Droga , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Malasia , Fibras de Estrés/metabolismo , Fibras de Estrés/patología
12.
Cell Prolif ; 52(6): e12689, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31502302

RESUMEN

OBJECTIVES: Osteogenesis is coupled with angiogenesis during bone remodelling. G-protein-coupled receptor (GPCR) kinase 2-interacting protein-1 (GIT1) is an important protein that participates in fracture healing by regulating angiogenesis. This study investigated whether GIT1 could affect bone mesenchymal stem cells (BMSCs) to secrete angiogenic factors to enhance fracture healing by promoting angiogenesis and its possible mechanism. MATERIALS AND METHODS: The angiogenesis of mice post-fracture was detected by micro-CT and immunofluorescence. Subsequently, vascular endothelial growth factor (VEGF) level in mouse and human BMSCs (hBMSCs) under TNF-α stimulation was detected. The hBMSCs were transfected with GIT1 shRNAs to further explore the relationship between GIT1 and VEGF and angiogenesis in vitro. Furthermore, based on previous research on GIT1, possible signal pathways were investigated. RESULTS: GIT1 knockout mice exhibited impaired angiogenesis and delayed fracture healing. And GIT1 deficiency remarkably reduced the expression of VEGF mRNA in BMSCs, which affected the proliferation and migration of human umbilical vein endothelial cells. GIT1 knockdown inhibited the activation of Notch and NF-κB signals by decreasing nuclear transportation of NICD and P65/P50, respectively. Overexpression of the canonical NF-κB subunits P65 and P50 markedly increased NICD-dependent activation of recombination signal-binding protein-jκ reporter. Finally, GIT1 enhanced the affinity of NF-κB essential modulator (NEMO) for K63-linked ubiquitin chains via interaction with NEMO coiled-coil 2 domains. CONCLUSION: These data revealed a positive role for GIT1 by modulating the Notch/NF-κB signals which promoting paracrine of BMSCs to enhance angiogenesis and fracture healing.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas de Ciclo Celular/metabolismo , Células Madre Mesenquimatosas/metabolismo , Inductores de la Angiogénesis/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , FN-kappa B/metabolismo , Neovascularización Patológica/metabolismo , Receptores Notch/metabolismo , Transducción de Señal/fisiología
13.
ACS Appl Mater Interfaces ; 11(39): 36141-36153, 2019 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-31503444

RESUMEN

Wound treatment is a long-lasting clinical issue. Poor angiogenesis leading to delayed wound closure causes huge challenges for healing. Functional electrospun membranes have been established as an efficient strategy to promote wound recovery by protecting and improving vascular regeneration. Here, we aimed to investigate the effect of tazarotene, an active drug for angiogenesis, loaded in aligned electrospun nanofibrous barrier on a soft tissue wound. This aligned membrane was arranged in a single direction, and tazarotene could be released from its nanofibers sustainably. The in vitro study demonstrated that compared with the random drug-loaded or other control groups, the aligned tazarotene-loaded membranes [poly-caprolactone (PCL)/AT] could stimulate proliferation, migration, angiogenesis, and vascular endothelial growth factor secretion and its gene expression of human umbilical vein endothelial cells. Furthermore, the in vivo model showed that the prepared tazarotene-loaded aligned membrane significantly accelerated the speed of healing, improved the neovascularization and re-epithelialization, and inhibited the inflammatory reaction in the wound area. All these results above indicated that the PCL/AT nanofibrous dressing, which could promote angiogenesis because of both stimulation of structure and chemical signals, is a promising wound-caring material.


Asunto(s)
Sistemas de Liberación de Medicamentos , Membranas Artificiales , Neovascularización Fisiológica/efectos de los fármacos , Ácidos Nicotínicos , Piel , Cicatrización de Heridas/efectos de los fármacos , Animales , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Masculino , Ácidos Nicotínicos/química , Ácidos Nicotínicos/farmacología , Ratas , Ratas Sprague-Dawley , Piel/lesiones , Piel/metabolismo , Piel/patología
14.
Int J Nanomedicine ; 14: 5875-5894, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31534329

RESUMEN

Background: Theranostics based on multifunctional nanoparticles (NPs) is a promising field that combines therapeutic and diagnostic functionalities into a single nanoparticle system. However, the major challenges that lie ahead are how to achieve accurate early diagnosis and how to develop efficient and noninvasive treatment. Sonodynamic therapy (SDT) utilizing ultrasound combined with a sonosensitizer represents a novel noninvasive modality for cancer therapy. Different ultrasound frequencies have been used for SDT, nevertheless, whether the effect of SDT can enhance synergistic HIFU ablation remains to be investigated. Materials and methods: We prepared a nanosystem for codelivery of a sonosensitizer (methylene blue, MB) and a magnetic resonance contrast agent (gadodiamide, Gd-DTPA-BMA) based on hydrophilic biodegradable polymeric NPs composed of poly (lactic-co-glycolic acid) (PLGA). To enhance accumulation and penetration of the NPs at the tumor site, the surface of PLGA NPs was decorated with a tumor-homing and penetrating peptide-F3 and polyethylene glycol (PEG). The physicochemical, imaging and therapeutic properties of F3-PLGA@MB/Gd and drug safety were thoroughly evaluated both in vitro and in vivo. F3-PLGA@MB/Gd was evaluated by both photoacoustic and resonance imaging. Results: F3-PLGA@MB/Gd NPs exhibited higher cellular association than non-targeted NPs and showed a more preferential enrichment at the tumor site. Furthermore, with good drug safety, the apoptosis triggered by ultrasound in the F3-PLGA@MB/Gd group was greater than that in the contrast group. Conclusion: F3-PLGA@MB/Gd can work as a highly efficient theranostic agent, and the incorporation of targeted multimodal and combined therapy could be an encouraging strategy for cancer treatment.


Asunto(s)
Péptidos de Penetración Celular/química , Tratamiento con Ondas de Choque Extracorpóreas , Nanopartículas/química , Neoplasias/diagnóstico por imagen , Ultrasonografía , Animales , Bovinos , Muerte Celular , Línea Celular Tumoral , Terapia Combinada , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Imagen por Resonancia Magnética , Ratones Endogámicos BALB C , Ratones Desnudos , Nanopartículas/ultraestructura , Especies Reactivas de Oxígeno/metabolismo
15.
Mol Med Rep ; 20(3): 2945-2953, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31524245

RESUMEN

Lipopolysaccharide (LPS) can regulate the expression of apoptotic factors, including caspase­3, Bcl­2 and Bcl­2­associated X protein (Bax). Nitric oxide (NO) plays an important role in apoptosis. N­acetyl cysteine (NAC) has been shown to exhibit antioxidant effects in vitro. However, the effects of NAC on LPS­induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the associated mechanisms are not well characterized. The present study explored the effect of NAC on LPS­induced apoptosis of HUVECs and determined the participation of the p38 mitogen­activated protein kinase (MAPK) pathway in the process of apoptosis. Cell viability was assessed using the Cell Counting Kit­8 (CCK­8) assay. The expression of caspase­3, Bax, Bcl­2, phosphorylated (p)­p38MAPK/total (t­)p38MAPK and p­endothelial e nitric oxide synthase (eNOS)/t­eNOS proteins were determined by western blotting. The expression levels of caspase­3, Bax and Bcl­2 mRNA were determined using reverse transcription­quantitative polymerase chain reaction (RT­qPCR). The rate of apoptosis was determined using flow cytometry. An NO detection kit (nitric reductase method) was used to determine NO concentration. The results of CCK­8 and flow cytometric analyses showed that pretreatment of HUVECs with NAC or p38MAPK inhibitor (SB203580) attenuated LPS­induced decrease in cell viability and increase in cell apoptosis. RT­qPCR and western blotting showed that LPS promoted caspase­3 and Bax expression, but inhibited that of Bcl­2 in HUVECs; however, these effects were attenuated by pretreatment with NAC or SB203580. LPS stimulation significantly enhanced the expression of p­p38MAPK protein and reduced the expression of p­eNOS protein; however, these effects were attenuated by pretreatment with NAC or SB203580. NAC pretreatment attenuated LPS­induced inhibition of NO synthesis, which was consistent with the effects of SB203580. The results demonstrated that NAC pretreatment alleviated LPS­induced apoptosis and inhibition of NO production in HUVECs. Furthermore, these effects were proposed to be mediated via the p38MAPK signaling pathway.


Asunto(s)
Acetilcisteína/farmacología , Apoptosis/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Lipopolisacáridos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Biomarcadores , Supervivencia Celular/efectos de los fármacos , Humanos , Óxido Nítrico/metabolismo , Fosforilación/efectos de los fármacos
16.
Mol Med Rep ; 20(3): 2796-2804, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31524252

RESUMEN

The renin­angiotensin system (RAS) serves an essential role in hypertension. MicroRNAs (miRs) have been reported to be important regulators in angiotensin (Ang) II­dependent hypertension. We aimed to explore the roles of Ang II and miR­133a in the mechanism underlying hypertension. Human umbilical vein endothelial cells (HUVECs) were identified by immunofluorescence staining. Cell viability and miR­133a expression under the inhibition of Ang II of various concentrations were determined by an MTT assay and reverse transcription­quantitative polymerase chain reaction (RT­qPCR), respectively. The effects of HUVECs transfected with miR­133a mimic or inhibitor on Ang II­induced apoptosis were measured using flow cytometry. The potential targeting of miR­133a to the 3' untranslated region of (pro) renin receptor (PRR) was assessed using TargetScan and a dual­luciferase assay. The effects of PRR interference using small interfering (si)RNA on PRR expression and the rate of apoptosis were determined by RT­qPCR, western blotting and flow cytometry, respectively. Ang II at a concentration of 10­5 M significantly inhibited the cell viability (P<0.05) and miR­133a expression (P<0.01); Downregulation of miR­133a suppressed cell viability. HUVECs transfected with miR­133a mimic reduced the rate of Ang II­induced apoptosis from 21.99 to 12.38%, but miR­133a inhibitor promoted Ang II­induced apoptosis (apoptosis rate, 28.9%). PRR was predicted to be a target gene of miR­133a. Transfection with siPRR decreased the apoptotic rate in Ang II + negative control and Ang II + miR­133a inhibitor group to 11.39 and 12.94%, respectively. Our findings also suggested that Ang II promoted PRR expression to enhance the apoptotic rate of HUVECs via the suppression of miR­133a. Furthermore, siPRR efficiently decreased the Ang II­induced apoptosis.


Asunto(s)
Regulación de la Expresión Génica , Células Endoteliales de la Vena Umbilical Humana/metabolismo , MicroARNs/genética , Interferencia de ARN , Regiones no Traducidas 3' , Angiotensina II/metabolismo , Angiotensina II/farmacología , Biomarcadores , Supervivencia Celular/efectos de los fármacos , Técnica del Anticuerpo Fluorescente , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Hipertensión/etiología , Hipertensión/metabolismo , Hipertensión/fisiopatología , Receptores de Superficie Celular/metabolismo , Sistema Renina-Angiotensina , ATPasas de Translocación de Protón Vacuolares/metabolismo
17.
Life Sci ; 239: 116874, 2019 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-31521690

RESUMEN

Atherosclerosis (AS) is a chronic inflammatory disease that results from Oxidized low-density lipoprotein (Ox-LDL) induced endothelial dysfunction. Cytoplasmic polyadenylation element binding protein 1 (CPEB1) is closely related to the development of epithelial cells, but the role of CPEB1 in AS remains unknown. The RNA and protein levels of CPEB1 expression are increased by Ox-LDL exposure, which is abrogated by c-Jun amino-terminal kinase (JNK) inhibitor SP600125. CPEB1 small interfering RNA (siRNA) suppressed the oxidative stress, inflammation, and apoptosis. Furthermore, CPEB1 siRNA enhanced the sirtuin 1 (SIRT1) transcription levels in Ox-LDL-treated HUVECs. Co-Immunoprecipitation (Co-IP) assay showed that CPEB1 siRNA declined the ubiquitination of SIRT1, and SIRT1 siRNA enhanced the Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), which were decreased by CPEB1 siRNA. In addition, LOX-1 and SIRT1 attenuated the protection of SIRT1 siRNA on Ox-LDL-induced oxidative stress. Therefore, our study revealed that CPEB1 depletion might play an anti-inflammatory and antiapoptotic role in Ox-LDL-induced apoptosis and inflammation though SIRT1/LOX-1 signalling pathway.


Asunto(s)
Receptores Depuradores de Clase E/metabolismo , Sirtuina 1/metabolismo , Factores de Transcripción/deficiencia , Factores de Escisión y Poliadenilación de ARNm/deficiencia , Apoptosis/fisiología , Aterosclerosis/metabolismo , Aterosclerosis/fisiopatología , Endotelio/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Inflamación/metabolismo , Lipoproteínas LDL/metabolismo , Estrés Oxidativo/efectos de los fármacos , Transducción de Señal , Factores de Transcripción/metabolismo , Factores de Escisión y Poliadenilación de ARNm/metabolismo
18.
Iran J Allergy Asthma Immunol ; 18(3): 281-288, 2019 Jun 08.
Artículo en Inglés | MEDLINE | ID: mdl-31522435

RESUMEN

Angiogenesis has essential role in growth and metastasis of tumors. Development of therapies aimed to suppress angiogenesis using medicinal plants is one of the effective approaches for prevention/treatment of cancer. The current study was performed to investigate in vitro anti-angiogenic effect of Teucrium Polium (TP) extract and its fractions. The aerial part of Teucrium Polium was powdered and extracted with 50% ethanol. The extract was fractionated in to aqueous (AQ), n-butanol (BU), ethyl acetate (EA) and n-hexane (HE) fractions. Anti-angiogenic effect of TP was examined on human umbilical vein endothelial cells (HUVECs) in three-dimensional collagen matrix. The endothelial cells form capillary-like branches that can be visualized using phase contrast microscope and the number of tube-like structures can be quantified as a measure of in vitro angiogenesis. Furthermore, anti-proliferative and vascular endothelial growth factor(VEGF )suppressive effect of TP as important factors in the process of angiogenesis were assessed using3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)and quantitative ELISA, respectively. Based on our findings, among the TP fractions, EA fraction showed the highest inhibitory activity on angiogenesis. This fraction with IC50: 68 µg/mL, inhibited angiogenesis at 60 µg/mL. The crude extract and other fractions of TP inhibited angiogenesis in a dose-dependent manner at doses higher concentrations than EA fraction, significantly.TP extract and EA fraction were able to inhibit proliferation of HUVEC and inhibited VEGF secretion in a dose dependent manner. The ethyl acetate fraction at 60 µg/ml inhibited VEGF secretion perfectly. Our data indicated that ethyl acetate fraction of Teucrium Polium could be a potential candidate for the prevention of angiogenesis in cancer and other related disorders. However, this suggestion needs more quantitative and in vivo investigations for confirmation.


Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Colágeno/metabolismo , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Neovascularización Fisiológica/efectos de los fármacos , Extractos Vegetales/farmacología , Teucrium/química , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Inhibidores de la Angiogénesis/química , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Humanos , Extractos Vegetales/química
19.
Nanoscale ; 11(38): 17699-17708, 2019 Oct 03.
Artículo en Inglés | MEDLINE | ID: mdl-31545331

RESUMEN

Biomaterial topography-based strategies are regarded as an effective way to regulate the osteoimmune environment which plays an indispensable role in the bone regeneration process. The rapid development of manufacture techniques makes it possible to investigate the cell-topography interactions by preparing various micro and nano-topographical surfaces on biomaterials. Still, it is a challenge to prepare well-defined micro/nano hierarchical structures of bioceramics due to the inherent brittleness of ceramic materials. Also, the correlation between osteoimmunomodulation initiated by micro/nano hierarchical topographies and the tissue regeneration outcomes is unclear. In this study, we prepared well-defined micro/nano hierarchical structures on hydroxyapatite (HA) bioceramics through the combination of the photolithography and hydrothermal techniques. Three different microscale circular patterns (4 µm, 12 µm and 36 µm) and nanotopographies (nanoneedle, nanosheet and nanorod) were fabricated by changing the size of the mask and the condition of the hydrothermal reaction. The macrophage responses on the nanoneedle structures with different micropatterns were investigated and the micro/nano hierarchical structures with appropriate pattern sizes could either promote or alleviate the macrophage polarization, which further affected the outcomes of the osteogenic differentiation of human bone marrow stromal cells (hBMSCs) and angiogenic activity of human umbilical vein endothelial cells (HUVECs). Our study demonstrated that osteoimmunomodulation could be manipulated via tuning the micro/nano hierarchical structures, which could lead to a new strategy for the development of bone biomaterials with favorable osteoimmunomodulatory properties.


Asunto(s)
Células de la Médula Ósea/metabolismo , Cerámica , Durapatita , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Nanopartículas/química , Neovascularización Fisiológica/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Animales , Células de la Médula Ósea/citología , Cerámica/química , Cerámica/farmacología , Durapatita/química , Durapatita/farmacología , Células Endoteliales de la Vena Umbilical Humana/citología , Humanos , Ratones , Células RAW 264.7 , Células del Estroma/citología , Células del Estroma/metabolismo
20.
Mol Med Rep ; 20(5): 4349-4357, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31545445

RESUMEN

The purpose of the study was to explore the potential protective effects of N­acetylcysteine (NAC) against lipopolysaccharide (LPS)­induced inflammatory injury to human umbilical vein endothelial cells (HUVECs). It was also assessed whether the underlying mechanism of this protective effect is mediated via suppression of the nuclear factor­kappa B (NF­κB) signaling pathway. Cell viability of HUVECs treated with different concentrations of NAC was assessed using Cell Counting Kit­8 (CCK­8) assay. The mRNA expression of inflammatory factors [interleukin­8 (IL­8), tumor necrosis factor α (TNF­α), inducible nitric oxide synthase (iNOS), and intercellular cell adhesive molecule 1 (ICAM­1)] were assessed using real time semi­quantitative polymerase chain reaction. Protein expression levels of TNF­α and IL­8 were assessed using enzyme­linked immunosorbent assay. Protein expression levels of ICAM­1 and the NF­κB signaling pathway were assessed using western blotting. Nitric reductase method was used to quantify nitric oxide (NO) and iNOS. LPS stimulated the production of TNF­α, IL­8, NO, and ICAM­1 by HUVECs. Moreover, LPS induced activation of the NF­κB signaling pathway and increased the protein expression of phosphorylated p65. However, pretreatment of HUVECs with NAC significantly attenuated the increase in the expression of inflammatory factors and the level of phosphorylated p65; this indicated that NAC prevented the activation of the NF­κB signaling pathway. The present findings indicated that NAC protects HUVECs against LPS­mediated inflammatory reaction and alleviates inflammation. The underlying mechanism is related to the NF­κB signaling pathway. NAC appears to be a promising agent for prevention and treatment of inflammatory diseases.


Asunto(s)
Acetilcisteína/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Biomarcadores , Supervivencia Celular/efectos de los fármacos , Citocinas/metabolismo , Humanos , Inflamación/etiología , Inflamación/metabolismo , Mediadores de Inflamación/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Lipopolisacáridos/efectos adversos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo
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