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INTRODUCTION: The Enhanced Control of Hypertension and Thrombolysis Stroke Study (ENCHANTED) showed that a low-dose alteplase was safe but not clearly non-inferior to standard-dose alteplase in acute ischemic stroke (AIS). Given the significant cost of this medicine, we undertook a cost-effectiveness analysis to determine the probability that low-dose is cost-effective relative to standard-dose alteplase in China. METHODS: For ENCHANTED participants in China with available health cost data, cost-effectiveness and cost-utility analyses were undertaken in which death or disability (modified Rankin scale scores 2-6) at 90 days and quality-adjusted life-years (QALYs) were used as outcome measures, respectively. There was adherence to standard guidelines for health economic evaluations alongside non-inferiority trials and according to a health-care payer's perspective. The equivalence margin for cost and effectiveness was set at USD 691 and -0.025 QALYs, respectively, for the base-case analysis. Probabilistic sensitivity analyses were used to evaluate the probability of low-dose alteplase being non-inferior. RESULTS: While the mean cost of alteplase was lower in the low-dose group (USD 1,569 vs. USD 2,154 in the standard-dose group), the total cost was USD 56 (95% confidence interval [CI]: -1,000-1,113) higher compared to the standard-dose group due to higher hospitalization costs in the low-dose group. There were 462 (95% CI: 415-509) and 410 (95% CI: 363-457) patients with death or disability per 1,000 patients in the low-dose and standard-dose groups, respectively. The low-dose group had marginally lower (0.008, 95% CI: -0.016-0.001) QALYs compared to their standard-dose counterparts. The low-dose group was found to have an 88% probability of being non-inferior based on cost-effectiveness versus the standard-dose group. CONCLUSIONS: This health economic evaluation alongside the ENCHANTED indicates that the use of low-dose alteplase does not save overall healthcare costs nor lead to a gain in QALYs in the management of Chinese patients with AIS compared to the use of standard dose. There is little justification on economic grounds to shift from standard-of-care thrombolysis in AIS.
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Isquemia Encefálica , Accidente Cerebrovascular Isquémico , Accidente Cerebrovascular , Humanos , Activador de Tejido Plasminógeno/efectos adversos , Análisis Costo-Beneficio , Fibrinolíticos/efectos adversos , Accidente Cerebrovascular Isquémico/tratamiento farmacológico , China , Resultado del TratamientoRESUMEN
BACKGROUND: To compare neurocognitive functioning of Type 1 diabetic mellitus (T1DM) and healthy adults, and explore risk factors of cognitive dysfunction of T1DM patients, especially the association between cognitive impairment and diabetic peripheral neuropathy (DPN). METHODS: Seventy T1DM (age: 32.17 ± 9.57 yr., duration: 8.99 ± 7.02 yr) patients and 48 healthy volunteers were included. All subjects received evaluation of MMSE and MoCA scales. Cognitive function of T1DM patients was evaluated in different cognitive domains. Risk factors affecting cognitive function were further explored. RESULTS: Three patients with educational level ≤ 6-year were excluded from final analysis. Scores of both MMSE (28.4 ± 1.7 vs. 29.1 ± 1.0, P = 0.005) and MoCA scales (25.9 ± 2.7 vs.27.1 ± 2.4, P = 0.017) in T1DM group were lower than that in control group. For MMSE scale, scores of orientation (9.60 ± 0.79 vs.9.87 ± 0.39, P < 0.001) and language function (8.56 ± 0.65 vs.8.83 ± 0.38, P < 0.001) in T1DM groups were lower than that in control group. For MoCA scale, scores of attention and concentration (2.30 ± 0.74 vs.2.57 ± 0.58, P < 0.001), visuospatial/executive function (4.32 ± 0.91 vs.4.64 ± 0.63, P < 0.001), memory (2.96 ± 1.50 vs.3.66 ± 1.28, P < 0.001), language function (5.71 ± 0.69 vs.5.87 ± 0.39, P = 0.007), and abstraction (1.55 ± 0.68 vs.1.82 ± 0.42, P < 0.001) were lower in T1DM group than that in control group. Logistic regression showed age, fasting C peptide, educational level and nerve conduction velocity (NCV) were associated with cognitive dysfunction diagnosed by MoCA scores for the patients with type 1 diabetes. CONCLUSIONS: T1DM adults had mild to moderate cognitive impairment, mainly presenting as dysfunctions of attention and concentration, visuospatial/executive, language, and abstraction. In addition to age, fasting C peptide level, and educational level, DPN, as a diabetic complication, was identified to be associated with cognitive impairments.
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Disfunción Cognitiva/etiología , Diabetes Mellitus Tipo 1/complicaciones , Neuropatías Diabéticas/etiología , Adulto , Estudios de Casos y Controles , China , Disfunción Cognitiva/patología , Estudios Transversales , Neuropatías Diabéticas/patología , Femenino , Estudios de Seguimiento , Humanos , Masculino , Pronóstico , Estudios Prospectivos , Factores de RiesgoRESUMEN
Rapamycin and its derivative possess anti-atherosclerosis activity, but its effects on adhesion molecule expression and macrophage adhesion to endothelial cells during atherosclerosis remain unclear. In this study we explored the effects of rapamycin on ox-LDL-induced adhesion molecule expression and macrophage adhesion to endothelial cells in vitro and the underlying mechanisms. Ox-LDL (6-48 µg/mL) dose-dependently increased the protein levels of two adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and E-selectin, in human umbilical vein endothelial cells (HUVECs), whereas pretreatment with rapamycin (1-10 µmol/L) dose-dependently inhibited ox-LDL-induced increase in the adhesion molecule expression and macrophage adhesion to endothelial cells. Knockdown of mTOR or rictor, rather than raptor, mimicked the effects of rapamycin. Ox-LDL (100 µg/mL) time-dependently increased PKC phosphorylation in HUVECs, which was abolished by rapamycin or rictor siRNA. Pretreatment with PKC inhibitor staurosporine significantly reduced ox-LDL-stimulated adhesion molecule expression and macrophage adhesion to endothelial cells, whereas pretreatment with PKC activator PMA/TPA attenuated the inhibitory effect of rapamycin on adhesion molecule expression. Ox-LDL (100 µg/mL) time-dependently increased c-Fos levels in HUVECs, and pretreatment with rapamycin or rictor siRNA significantly decreased expression of c-Fos. Knockdown of c-Fos antagonized ox-LDL-induced adhesion molecule expression and macrophage adhesion to endothelial cells. Our results demonstrate that rapamycin reduces ox-LDL-stimulated adhesion molecule expression and macrophage adhesion to endothelial cells by inhibiting mTORC2, but not mTORC1, and mTORC2 acts through the PKC/c-Fos signaling pathway.
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Genes fos/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Inflamación/prevención & control , Lipoproteínas LDL/antagonistas & inhibidores , Diana Mecanicista del Complejo 2 de la Rapamicina/antagonistas & inhibidores , Proteína Quinasa C/antagonistas & inhibidores , Sirolimus/farmacología , Adhesión Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Selectina E/metabolismo , Técnicas de Silenciamiento del Gen , Células Endoteliales de la Vena Umbilical Humana/enzimología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Lipoproteínas LDL/farmacología , Diana Mecanicista del Complejo 2 de la Rapamicina/genética , ARN Interferente Pequeño/farmacología , Proteína Asociada al mTOR Insensible a la Rapamicina/antagonistas & inhibidores , Proteína Asociada al mTOR Insensible a la Rapamicina/genética , Proteína Reguladora Asociada a mTOR/antagonistas & inhibidores , Proteína Reguladora Asociada a mTOR/genética , Transducción de Señal/efectos de los fármacos , Estaurosporina/farmacología , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Foam cell formation, which is caused by imbalanced cholesterol influx and efflux by macrophages, plays a vital role in the occurrence and development of atherosclerosis. Humanin (HN), a mitochondria-derived peptide, can prevent the production of reactive oxygen species and death of human aortic endothelial cells exposed to oxidized low-density lipoprotein (ox-LDL) and has a protective effect on patients with in early atherosclerosis. However, the effects of HN on the regulation of cholesterol metabolism in RAW 264.7 macrophages are still unknown. This study was designed to investigate the role of [Gly14]-humanin (HNG) in lipid uptake and cholesterol efflux in RAW 264.7 macrophages. Flow cytometry and live cell imaging results showed that HNG reduced Dil-ox-LDL accumulation in the RAW 264.7 macrophages. A similar result was obtained for lipid accumulation by measuring cellular cholesterol content. Western blot analysis showed that ox-LDL treatment upregulated not only the protein expression of CD36 and LOX-1, which mediate ox-LDL endocytosis, but also ATP-binding cassette (ABC) transporter A1 and ABCG1, which mediate ox-LDL exflux. HNG pretreatment inhibited the upregulation of CD36 and LOX-1 levels, prompting the upregulation of ABCA1 and ABCG1 levels induced by ox-LDL. Therefore we concluded that HNG could inhibit ox-LDL-induced macrophage-derived foam cell formation, which occurs because of a decrease in lipid uptake and an increase in cholesterol efflux from macrophage cells.
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Diferenciación Celular/fisiología , Colesterol/metabolismo , Células Espumosas/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Metabolismo de los Lípidos/fisiología , Lipoproteínas LDL/farmacocinética , Animales , Células Espumosas/citología , Ratones , Células RAW 264.7RESUMEN
Recent studies suggest that epigenetic alterations such as DNA methylation control many aspects of monocytes/macrophages and participate in the pathogenesis of atherosclerosis, a lipid-driven inflammatory disorder. Our and other groups demonstrated that dysregulation of cystathionine γ-lyase (CSE) -hydrogen sulfide (H2S) pathway was involved in monocyte/macrophages-mediated inflammation and atherosclerosis. However, it remains unknown whether altered cse methylation in macrophages may play a role in linking CSE-H2S dysregulation and atherosclerosis. In the present study, we showed that plasma H2S and H2S production in the peritoneal macrophages of apolipoprotein knockout (apoE(-/-)) mice gradually decreased with ages, and were also lower than that in control mice at 12 weeks older. Moreover, CSE mRNA expressions decreased while DNA methyltransferase (DNMT) expressions increased in the peritoneal macrophages isolated from apoE(-/-) mice, compared to age-matched wildtype mice. Similar observations were obtained in an in vitro study. In oxidized low-density lipoprotein (ox-LDL)-treated raw264.7 macrophages, cse transcription was down-regulated while the expression and activity of DNMT was up-regulated, associated with enhanced DNA methylation in cse promoter. Suppression of DNMT with its inhibitor or siRNA reversed the decrease of CSE mRNA. Therefore, our data suggest that DNA hypermethylation of CpG rich region in cse promoter might contribute to the decrease of cse transcription and H2S production in macrophages, and thus contribute to atherosclerosis development.
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Cistationina gamma-Liasa/genética , Metilación de ADN/genética , Sulfuro de Hidrógeno/sangre , Lipoproteínas LDL/farmacología , Macrófagos/fisiología , Regiones Promotoras Genéticas/genética , Animales , Células Cultivadas , Metilación de ADN/efectos de los fármacos , Macrófagos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Células RAW 264.7RESUMEN
Hyperhomocysteinemia (HHcy) is an independent risk factor of atherosclerosis and other cardiovascular diseases. Unfortunately, Hcy-lowering strategies were found to have limited effects in reducing cardiovascular events. The underlying mechanisms remain unclear. Increasing evidence reveals a role of inflammation in the pathogenesis of HHcy. Homocysteine (Hcy) is a precursor of hydrogen sulfide (H2S), which is formed via the transsulfuration pathway catalyzed by cystathionine ß-synthase and cystathionine γ-lyase (CSE) and serves as a novel modulator of inflammation. In the present study, we showed that methionine supplementation induced mild HHcy in mice, associated with the elevations of TNF-α and IL-1ß in the plasma and reductions of plasma H2S level and CSE expression in the peritoneal macrophages. H2S-releasing compound GYY4137 attenuated the increases of TNF-α and IL-1ß in the plasma of HHcy mice and Hcy-treated raw264.7 cells while CSE inhibitor PAG exacerbated it. Moreover, the in vitro study showed that Hcy inhibited CSE expression and H2S production in macrophages, accompanied by the increases of DNA methyltransferase (DNMT) expression and DNA hypermethylation in cse promoter region. DNMT inhibition or knockdown reversed the decrease of CSE transcription induced by Hcy in macrophages. In sum, our findings demonstrate that Hcy may trigger inflammation through inhibiting CSE-H2S signaling, associated with increased promoter DNA methylation and transcriptional repression of cse in macrophages.
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Cistationina gamma-Liasa/genética , Metilación de ADN/efectos de los fármacos , Homocisteína/farmacología , Hiperhomocisteinemia/inducido químicamente , Mediadores de Inflamación/farmacología , Macrófagos/efectos de los fármacos , Animales , Células Cultivadas , Cistationina gamma-Liasa/metabolismo , ADN (Citosina-5-)-Metiltransferasas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Sulfuro de Hidrógeno/metabolismo , Hiperhomocisteinemia/inmunología , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Metionina/administración & dosificación , Metionina/efectos adversos , Ratones , Morfolinas/farmacología , Compuestos Organotiofosforados/farmacología , Regiones Promotoras Genéticas/efectos de los fármacos , Transducción de Señal/efectos de los fármacosRESUMEN
Hydrogen sulfide (H2S), the third gaseous transmitter, is implicated in various pathophysiologic processes. In the cardiovascular system, H2S exerts effects of cardioprotection, vascular tone regulation, and atherogenesis inhibition. Recent studies demonstrated that atorvastatin, the inhibitor of 3-hydroxyl-3-methyl coenzyme A reductase, affected H2S formation in kidney and other organs. However, the underlying mechanisms are not fully understood. In this study, we examined the effects of three different statins (fluvastatin, atorvastatin and pravastatin) on H2S formation in raw264.7 macrophages. There was a remarkable rise in H2S level in fluvastatin- and atorvastatin-stimulated macrophages, while pravastatin failed to show any significant effect on it. Moreover, fluvastatin and atorvastatin enhanced the mRNA and protein expression of cystathionine γ-lyase (CSE) in dose- and time-dependent manners. Fluvastatin also markedly enhanced the CSE activity. However, fluvastatin did not alter the mRNA or protein expression of another H2S-producing enzyme 3-mercaptopyruvate sulfurtransferase. Blockade of CSE with its inhibitor dl-propargylglycine (PAG) or siRNA markedly reduced the H2S level in fluvastatin-stimulated macrophages. In addition, fluvastatin elevated Akt phosphorylation, which occurred as early as 15 min after treatment, peaked at 1h, and lasted at least 3h. Both PI3K inhibitor LY294002 (10 µM) and Akt inhibitor perifosine (10µM) were able to reverse the increases of CSE mRNA and H2S production in fluvastatin-stimulated macrophages. Last, we showed that fluvastatin reduced the mRNA levels of pro-inflammatory molecules such as IL-1ß and MCP-1 in LPS-treated macrophages, which were completely reversed by CSE inhibitor PAG. Taken together, the findings demonstrate that statins may up-regulate CSE expression/activity and subsequently elevate H2S generation by activating Akt signaling pathway and also imply that CSE-H2S pathway plays a critical role in the anti-inflammation elicited by statins.
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Cistationina gamma-Liasa/metabolismo , Sulfuro de Hidrógeno/metabolismo , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Macrófagos/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Atorvastatina , Línea Celular , Cistationina gamma-Liasa/genética , Ácidos Grasos Monoinsaturados/farmacología , Fluvastatina , Ácidos Heptanoicos/farmacología , Indoles/farmacología , Macrófagos/metabolismo , Ratones , Pravastatina/farmacología , Pirroles/farmacología , ARN Mensajero/metabolismo , Transducción de Señal , Regulación hacia ArribaRESUMEN
AIMS AND OBJECTIVES: To explore which sociodemographic and clinical factors could interfere in the parameters of ambulatory blood pressure monitoring and determine the affecting factors of Ambulatory Arterial Stiffness Index. BACKGROUND: Although the clinical relevance of ambulatory blood pressure monitoring and Ambulatory Arterial Stiffness Index have been studied, the explanation of their role and related interfering factors remains controversial in patients with different disease or age, etc. DESIGN: Cross-sectional study. METHODS: The study was carried out between October 2008-October 2009. A convenience sample of hypertensive patients over 60 years old was recruited in China. Twenty-four hour ambulatory blood pressure monitoring was carried out on the non-dominant arm using an oscillometric device. RESULTS: (1) All 95 patients completed the study and their ages ranged from 60-76 years. (2) There were statistical differences for certain parameters of ambulatory blood pressure monitoring between different characteristics of patients. Financial status was an important factor interfering in patients' BP fluctuation, especially daytime and 24 hours systolic pressure. The higher body mass index the patients had, the higher the pressure was. (3) Multiple variants logistic analysis of Ambulatory Arterial Stiffness Index showed statistical differences only in coefficient variation of 24-hour diastolic pressure and daytime systolic blood pressure. CONCLUSIONS: There are more factors that interfered with systolic pressure and diastolic pressure during the day than night. Patients who have less nocturnal dipping may have a higher night time systolic pressure and diastolic pressure. The coefficient of variation of 24 hours diastolic pressure and daytime systolic pressure contribute more to Ambulatory Arterial Stiffness Index which should arouse practitioners' attention. RELEVANCE TO CLINICAL PRACTICE: Ambulatory blood pressure monitoring should be used as a routine procedure, as well as Ambulatory Arterial Stiffness Index calculated for older hypertensive patients. The findings may be used to guide community health providers to pay more attention to the factors that may influence BP fluctuation and Ambulatory Arterial Stiffness Index according to individual's characteristics.
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Presión Sanguínea , Hipertensión/fisiopatología , Rigidez Vascular , Anciano , China , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Syphilis is now rare and easily misdiagnosed because of the wide use of antibiotics in the clinical. We report a case of cerebral hemorrhage in a patient with hypertension who was first diagnosed as hypertensive cerebral hemorrhage. However, treponema pallidum particle agglutination and rapid plasma regain tests of cerebrospinal fluid revealed the existence of neurosyphilis. Interestingly, digital subtraction angiography (DSA) showed severe stenosis in both middle cerebral arteries and right anterior cerebral artery. The case reminded us to pay attention to syphilitic vasculitis in patients with cryptogenic stroke. DSA sometimes may play a critical role in differential diagnosis of neurosyphilis.
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Angiografía de Substracción Digital , Hemorragia Cerebral/diagnóstico por imagen , Neurosífilis/diagnóstico por imagen , Vasculitis del Sistema Nervioso Central/diagnóstico por imagen , Angiografía de Substracción Digital/métodos , Hemorragia Cerebral/etiología , Femenino , Humanos , Persona de Mediana Edad , Neurosífilis/complicaciones , Vasculitis del Sistema Nervioso Central/complicacionesRESUMEN
Background: Endovascular treatment (EVT) is one of the effective treatment procedure for the symptomatic intracranial atherosclerotic stenosis (sICAS). Aim and methods: We evaluated the efficacy and safety of individualized endovascular treatment for sICAS patients. Clinical and imaging follow-ups were carried out to collect the data of 29 sICAS patients after 6 months of individualized endovascular treatment. Different treatment strategies are selected based on arterial access and lesion morphology of patients. If standard surgical path, narrow artery straight, stenosis length ≤10 mm, then the appropriate specifications of balloon-mounted stent (BMS) treatment. the surgical path is tortuous, the narrow artery is curved, the angle is apparent, the diameter of the near and far ends is significantly different, or the length of the stenosis is >10 mm, self-expanding stent (SES) with appropriate specifications is selected for treatment. If the narrowed artery is hyper flexed and the surgeon deems stenting inappropriate, balloon dilation angioplasty (BDA) treatment is chosen. Results and conclusion: 31 lesions of 29 sICAS patients received endovascular treatment. The median age was 61 years (IQR 54-69 years). The median preoperative stenosis was 90% (IQR 80-95%), and the mean stenosis length was (8.10 ± 3.27) mm. The most commonly used surgical procedure was Balloon-Mounted Stent (BMS) in 19 cases (65.52%), Self-expanding Stent (SES) in seven cases (24.14%), Balloon Dilation Angioplasty (BDA) in three cases (10.34%). (11.86 + 1.46 mm) was greater than that in the BMS group (6.14 + 1.59 mm) (P < 0.001). The median stenosis was 90% (IQR 80-92.5%) in the BMS group, lower than 99% (IQR 95-100%) in the SES group (P < 0.001). The median post-operative residual stenosis was 20% (IQR 15-25%), significantly improved compared with preoperative (P < 0.001). The success rate of the surgical technique was 93.10% (27/29). One patient (3.45%) had IS recurrence within 48 h after surgery, and the restenosis rate within 6 months after surgery was 6.90% (2/29). No patient died or had recurrent IS. Our data demonstrated that individualized endovascular treatment method could be potentially significant and safe for sICAS patients. This study will provide an important reference for the endovascular treatment of sICAD.
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Jian-Gan-Xiao-Zhi decoction (JGXZ) has demonstrated beneï¬cial eï¬ects on nonalcoholic fatty liver disease (NAFLD). However, the mechanisms by which JGXZ improve NAFLD are still unclear. Methods. In this study, we first used a high-fat diet (HFD) to establish a NAFLD rat model to clarify the therapeutic effect of JGXZ on NAFLD. Secondly, we used network pharmacology to predict the potential targets of JGXZ on NAFLD, and then the key targets obtained from network pharmacology were verified. Finally, we used untargeted metabolomics to study the metabolic regulatory mechanism of JGXZ. Results. JGXZ treatment could decrease body weight and ameliorate dyslipidemia in NAFLD model rats. H&E and oil red O staining indicated that JGXZ reduced steatosis and inï¬ltration of inï¬ammatory cells in the liver. In addition, network pharmacology research found that the potential targets of JGXZ on NAFLD pathway were mainly associated with improving oxidative stress, apoptosis, inflammation, lipid metabolism disorders, and insulin resistance. Further experimental verification confirmed that JGXZ could inhibit inflammation and improve oxidative stress, insulin resistance, and lipid metabolism disorders. Serum untargeted metabolomics analyses indicated that the JGXZ in the treatment of NAFLD may work through the linoleic acid metabolism, alpha-linolenic acid metabolism, tryptophan metabolism, and glycerophospholipid metabolism pathways. Conclusions. In conclusion, this study found that JGXZ has an ameliorative effect on NAFLD, and JGXZ alleviates the inflammatory response and oxidative stress and lipid metabolism disorders in NAFLD rats. The mechanism of action of JGXZ in the treatment of NAFLD may be related to the regulation of linoleic acid metabolism, tryptophan metabolism, alpha-linolenic acid metabolism, and glycerophospholipid metabolism.
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OBJECTIVE: Our previous studies found that 100 µg/ml oxidized low-density lipoprotein (ox-LDL) could up-regulate the autophagic level in human umbilical vein endothelial cells (HUVEC). The present study was conducted to observe the roles of oxidative stress and lectin-like oxidized low density lipoprotein-1 (LOX-1) in the ox-LDL-induced up-regulation of autophagy. METHODS: Prior to the ox-LDL exposure, LOX-1mAb, vitamin C and vitamin E were used to study the roles of LOX-1 and oxidative stress in the activation of autophagy. The contents of total-superoxide dismutase (T-SOD) and MDA (malondialdehyde) in the culture medium were detected with enzyme linked immunosorbent assay. Western blot was employed to detect the levels of autophagic marker microtubule-associated protein light chain 3 (MAP1-LC3)-II/LC3-I, beclin1 and lysosome associated membrane protein 2a (lamp2a). RESULTS: After the ox-LDL exposure, the down-regulated level of T-SOD [0.5 h (32.73 ± 1.09 vs 40.16 ± 1.28) U/ml, P < 0.01; 6 h (29.32 ± 1.56 vs 40.16 ± 1.28) U/ml, P < 0.01] and the up-regulated level of MDA [0.5 h (1.11 ± 0.04 vs 0.57 ± 0.05) nmol/ml, P < 0.01; 6 h (0.69 ± 0.03 vs 0.57 ± 0.05) nmol/ml, P < 0.05] in culture medium were also significant at 0.5 h and 6 h. The ox-LDL-induced increased ratio of LC3-II/LC3-I was reversed by the pretreatments of vitamin C and vitamin E (0.5 h, vitC: 3.11 ± 0.02 vs 4.31 ± 0.50, P < 0.05; vitE: 3.46 ± 0.19 vs 4.31 ± 0.50, P < 0.05; 6 h, vitC: 1.44 ± 0.05 vs 2.31 ± 0.16, P < 0.05), but not LOX-1mAb. LOX-1mAb decreased the ox-LDL-induced elevated level of lamp2a protein while vitamin C and vitamin E only inhibited the elevation of lamp2a at the timepoint of 6 h, but not 0.5 h. CONCLUSION: Oxidative stress, rather than LOX-1, plays an important role in the ox-LDL-induced up-regulation of autophagy in HUVEC. The formation of autolysosomes is associated with the LOX-1-mediated endocytosis of ox-LDL. Oxidative stress only plays a minor role in the formation of autolysosomes induced by the engulfed ox-LDL.
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Autofagia , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Lipoproteínas LDL/metabolismo , Estrés Oxidativo , Receptores de LDL Oxidadas/metabolismo , Línea Celular , Humanos , Lectinas de Plantas , Regulación hacia ArribaRESUMEN
Oxidized low-density lipoprotein (ox-LDL) is involved in the pathogenesis of atherosclerosis and atherosclerotic plaque rupture by promoting lipid accumulation, proinflammatory responses, and cell death. LDL is mainly oxidized in the subendothelial layer of the vascular wall and then can be taken up by vascular endothelial cells. However, little is known about the intracellular processing of the damaged LDL. Previous studies found that autophagy is involved in degrading oxidized proteins under oxidative stress conditions in Arabidopsis thaliana, while ox-LDL can activate autophagy in EA.hy926 endothelial cells, suggesting a possible role of autophagy in the degradation of ox-LDL by endothelial cells. The present study showed that ox-LDL aggregated in human umbilical vein endothelial cells (HUVECs) and brought about an increase in the formation of autophagosomes and autolysosomes. Ox-LDL-induced increase in the autophagic level was blocked by treatment with the autophagy inhibitor 3-methyladenine and increased by the autophagy inducer rapamycin, while the aggregation of Dil-labled ox-LDL was increased by 3-methyladenine and decreased by rapamycin. In addition, Dil-labeled ox-LDL colocalized with the autophagy marker MDC, microtubule-associated protein light chain 3 (MAP1-LC3), and lysosome-associated membrane protein 2a (lamp2a). HUVECs treated with Dil-labeled-ox-LDL showed a much greater degree of overlap of MAP1-LC3 and Lamp2a than control. The results suggest that ox-LDL activates the autophagic lysosome pathway in HUVECs through the LC3/beclin1 pathway, leading to the degradation of ox-LDL.
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Autofagia , Endotelio Vascular/metabolismo , Lipoproteínas LDL/metabolismo , Lisosomas/metabolismo , Células Cultivadas , Humanos , Sirolimus/farmacologíaRESUMEN
OBJECTIVE: To investigate the role of autophagy in oxidized low density lipoprotein (ox-LDL) induced injury of human umbilical vein endothelial cells (HUVEC). METHODS: The cultured HUVECs were randomly divided into four groups of control, ox-LDL, ox-LDL + rapamycin and ox + 3-methyladenine (3-MA). The cells were used to detect the ratio of LC3-II/LC3-I by Western blot while the proliferation and apoptosis of cells measured by MTT and flow cytometry. The lactate dehydrogenase (LDH) activity and endothelin-1 (ET-1) content in the supernatant were detected with enzyme linked immunosorbent assay. RESULTS: The Ox-LDL treatment up-regulated the ratio of LC3-II/LC3-I in HUVEC (P < 0.01). It increased the activity of LDH (P < 0.01)and content of ET-1 (P < 0.05) in the supernatant. Also it induced the proliferation (P = 0.028) and apoptosis (P < 0.05) of cells. The autophagic inducer rapamycin increased the up-regulation of autophagic level induced by ox-LDL, decreased the activity of LDH and content of ET-1 (P < 0.05) and inhibited the ox-LDL-induced proliferation of cells. Conversely, the autophagic inhibitor 3-MA decreased the elevation of LC3-II/LC3-I induced by ox-LDL (P < 0.01) and increased the cell apoptosis and death. CONCLUSION: Ox-LDL exposure can increase the secretion of LDH and ET-1 and induce the proliferation and apoptosis of cells so as to cause a harmful effect in the survival of HUVEC. The injury may be reduced by rapamycin, an autophagic inducer, and elevated by the autophagic inhibitor 3-MA.
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Apoptosis , Autofagia , Células Endoteliales/citología , Lipoproteínas LDL/efectos adversos , Células Cultivadas , Endotelina-1/metabolismo , Endotelio Vascular/citología , Endotelio Vascular/metabolismo , Humanos , Venas Umbilicales/citología , Venas Umbilicales/metabolismoRESUMEN
BACKGROUND: Studies have shown that platelet-leukocyte aggregates (PLA) are sensitive to platelet activation which might exist before the onset of cerebral infarction. In this study, we investigated the formation of PLA in patients with cerebral infarction and the effects of antiplatelet agents on PLA. METHODS: The level of soluble P-selectin, C-reaction protein, platelet aggregation rate and leukocyte-platelet aggregations were measured in 40 patients with acute cerebral infarction and 20 normal controls. The 40 patients were randomly assigned to two treatment groups: aspirin group (n = 20) and clopidogrel group (n = 20). Both groups were monitored for Scandinavian stroke scale (SNSS), soluble P-selectin, serum C-reaction protein, platelet aggregation rate and PLA before and after the treatment. Flow cytometry was used to detect the levels of PLA in the blood. RESULTS: The percentage of platelet-monocyte aggregates (PMA) in patients with cerebral infarction was significantly increased compared with the controls (P < 0.001), which was positively correlated to soluble P-selectin, C-reaction protein and platelet aggregation rate (P < 0.05). After the treatment, the levels of PMA and platelet aggregation rate were decreased in both groups (P < 0.05). The level of PMA and platelet aggregation rate in the clopidogrel group was significantly lower than that in the aspirin group (P < 0.05). CONCLUSIONS: PMA are a sensitive biomarker to platelet activation in patients with cerebral infarction. In addition, although both aspirin and clopidogrel lowered the level of PMA, clopidogrel is a more effective treatment than aspirin in inhibiting platelet activation.
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Plaquetas/patología , Adhesión Celular/efectos de los fármacos , Infarto Cerebral/tratamiento farmacológico , Leucocitos/patología , Anciano , Aspirina/farmacología , Clopidogrel , Femenino , Humanos , Masculino , Persona de Mediana Edad , Activación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/farmacología , Ticlopidina/análogos & derivados , Ticlopidina/farmacologíaRESUMEN
This study aimed to compare the clinical efficacy of stenting compared with standardized medical treatment in patients with moderate to severe vertebral artery origin stenosis (VAOS).Patients diagnosed with moderate to severe VAOS and indicated to undergo vertebral artery stenting were enrolled. Patients were divided into stenting group and standardized medical treatment group. All patients underwent transcranial Doppler (TCD) before and after treatment. Incidence of new cerebral infarction, transient ischemic attack (TIA), improvement of clinical symptoms, and National Institutes of Health Stroke Scale (NIHSS) score were observed.A total of 98 patients were enrolled. Vertebral artery stenting implant was accepted by 43 patients. Two weeks after treatment, the NIHSS score in the stenting group decreased significantly compared to that in the standardized medical treatment group. The modified Rankin Scale (mRS) score in the stenting group at three months was significantly lower than that in the medical treatment group (Pâ=â.044). The extent of vascular stenosis in the stent group decreased significantly (76.5â±â10.0% vs. 13.7â±â5.9%, tâ=â35.878, Pâ=â.000). The adverse events occurred in 9 (16.4%) patients in the medical treatment group and 5 (11.6%) in the stenting group (Pâ=â.506). There was one case with new cerebral infarction in the stenting group, whereas the medical treatment group showed 1 case with TIA and three with new cerebral infarction during follow-up after 3 months. The peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) of stenosis vertebral artery, and PSV of basilar artery were significantly higher in the stent group than those in the standardized medical group (Pâ<â.05).Stenting for VAOS, rather than standardized medical treatment, can effectively relieve vascular stenosis, alter vertebral-basilar artery hemodynamics, and improve neurological function, with low perioperative complications.
Asunto(s)
Hemodinámica/fisiología , Stents/efectos adversos , Arteria Vertebral/diagnóstico por imagen , Insuficiencia Vertebrobasilar/diagnóstico por imagen , Anciano , Constricción Patológica/patología , Femenino , Estudios de Seguimiento , Humanos , Incidencia , Ataque Isquémico Transitorio/epidemiología , Ataque Isquémico Transitorio/etiología , Masculino , Persona de Mediana Edad , Ensayos Clínicos Controlados no Aleatorios como Asunto , Pronóstico , Estudios Prospectivos , Accidente Cerebrovascular/epidemiología , Accidente Cerebrovascular/etiología , Ultrasonografía Doppler Transcraneal/métodos , Arteria Vertebral/patología , Insuficiencia Vertebrobasilar/mortalidad , Insuficiencia Vertebrobasilar/terapiaRESUMEN
Dioscin, a steroidal saponin isolated from Dioscorea nipponica Makino, has previously been shown to possess antiarthritic effects. However, the underlying mechanism is still elusive. Herein, we investigated the therapeutic effects of dioscin on collagen-induced arthritis (CIA) in DBA/1 mice and related mechanism. Cytokine production in CII-specific immune responses were measured by enzyme-linked immunosorbent assay (ELISA); Th17 cell-related gene expression, including IL-17A, ROR γτ and IL-23p19, were detected by qPCR analysis; Surface marker, T regulatory (Treg) cells and intracellular cytokines (IL-17A and IFN- γ ) were evaluated by flow cytometry. We performed Th17 cell differentiation assay in vitro. Results showed that, in vivo, dioscin treatment significantly reduced the severity of CIA, which was accompanied by decreased Th17 response, but not Th1 and Treg response; dioscin-treated mice also showed lower percentage of CD11b + Gr-1 + neutrophils; In vitro, dioscin treatment suppressed the differentiation of naive CD4 + T cells into Th17 cell and decreased IL-17A production. Collectively, our results indicate that dioscin exerts antiarthritic effects by inhibiting Th17 cell immune response.
Asunto(s)
Artritis/tratamiento farmacológico , Artritis/inmunología , Colágeno/efectos adversos , Dioscorea/química , Diosgenina/análogos & derivados , Fitoterapia , Células Th17/inmunología , Animales , Artritis/inducido químicamente , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Células Cultivadas , Citocinas/metabolismo , Diosgenina/administración & dosificación , Diosgenina/aislamiento & purificación , Diosgenina/farmacología , Modelos Animales de Enfermedad , Femenino , Masculino , Ratones Endogámicos BALB C , Ratones Endogámicos DBARESUMEN
OBJECTIVE: To investigate the effect of fibrinogen (Fg), fibrin (Fb) and fibrin (ogen) degradation products (FDPs) on tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) expressions of human umbilical vein endothelial cells (HUVECs) in coculture system. METHODS: Fg, Fb and FDPs at various concentrations (0, 0.5, 1.5, 3.0, 4.5 and 6.0 g/L) were added to the transwell coculture system of HUVECs and smooth muscle cells (SMCs) for 24 hours. The expressions of tPA and PAI-1 at mRNA level were examined by RT-PCR and tPA and PAI-1 protein and activity were detected by ELISA and substrate chromogenic assays. RESULTS: tPA expression was not affected by Fg. Fg at concentrations between 3.0 - 4.5 g/L significantly enhanced the mRNA expression, protein content and activity of PAI-1, while expression of PAI-1 was significantly inhibited by Fg at concentration of 6.0 g/L. Fb at concentrations between 3.0 - 4.5 g/L significantly up-regulated mRNA expression, increased protein content and down-regulated activity of tPA. Fb (1.5 - 4.5 g/L) also enhanced the mRNA expression, increased protein content and activity of PAI-1. FDPs at concentrations 3.0 - 6.0 g/L down-regulated the expression of tPA and FDPs at concentrations 1.5 - 6.0 g/L significantly enhanced PAI-1 mRNA expression. CONCLUSION: Fg, Fb and FDPs play important roles in the pathogenesis of atherosclerosis by modulating the expression of tPA and PAI-1 of endothelial and SMCs.
Asunto(s)
Células Endoteliales/metabolismo , Productos de Degradación de Fibrina-Fibrinógeno/metabolismo , Fibrina/metabolismo , Fibrinógeno/metabolismo , Inhibidor 1 de Activador Plasminogénico/metabolismo , Activador de Tejido Plasminógeno/metabolismo , Animales , Arteriosclerosis/metabolismo , Arteriosclerosis/patología , Células Cultivadas , Técnicas de Cocultivo , Humanos , ARN Mensajero/metabolismo , ConejosRESUMEN
OBJECTIVE: To investigate the changes of platelet-leukocyte aggregation (PLA) in patients with cerebral infarction and the effects of antiplatelet agents on it. METHODS: The levels of plasma soluble P-selectin (sP-sel), serum C-reactive protein (CRP), platelet aggregation rate (PAR) and PLA were measured in 40 patients with acute cerebral infarction and 20 cases of controls. The 40 patients with cerebral infarction were randomly divided into aspirin-treated group (20 cases) and clopidogrel-treated group (20 cases) scandinavian neurological stroke score (SNSS), plasma sP-sel, serum CRP, PAR and PLA were observed before and after the treatment. RESULTS: Compared with controls, the percentage of monocyte-platelet aggregation (PMA) was significantly increased in the patients (P < 0.001). The level of plasma sP-sel, serum CRP and PAR in the case group were significantly greater than those in the control group (P < 0.05). The level of PMA was positively related with the plasma sP-sel, serum CRP and PAR in cerebral infarction patients before treatment (P < 0.05), and it showed a significant inverse correlation between the percentage of PMA and the score of SNSS (P < 0.05). After treatment, the levels of PMA and PAR were decreased in both groups of cerebral infarction patients (P < or = 0.001), and the level of PMA and PAR (ADP) in clopidogrel-treated group was lower than those in aspirin-treated group (P < 0.05). However, the CRP in the patients was no significant difference before and after treatment. The level of sP-sel was no significant difference before and after treatment in aspirin-treated group, but it was significantly decreased after treatment in clopidogrel-treated group (P < 0.001). CONCLUSIONS: The levels of PMA in patients with cerebral infarction was significantly higher than in the controls. PMA may be a sensitive marker of platelet activation. Aspirin and Clopidogrel could decline the level of PMA in the patients with acute cerebral infarction, and Clopidogrel was more effective than aspirin in inhibiting the activation of platelet.
Asunto(s)
Infarto Cerebral/tratamiento farmacológico , Inhibidores de Agregación Plaquetaria/uso terapéutico , Anciano , Aspirina/farmacología , Aspirina/uso terapéutico , Proteína C-Reactiva/metabolismo , Infarto Cerebral/sangre , Infarto Cerebral/fisiopatología , Clopidogrel , Femenino , Humanos , Masculino , Persona de Mediana Edad , Selectina-P/sangre , Activación Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/farmacología , Ticlopidina/análogos & derivados , Ticlopidina/farmacología , Ticlopidina/uso terapéutico , Resultado del TratamientoRESUMEN
BACKGROUND: Small nerve fibers are more easily injured than large ones for diabetic peripheral neuropathy (DPN). The study investigated the characteristics and related risk factors of DPN of T1DM patients using nerve conduction velocity and CPT values, which provided evidences for its early diagnosis. METHODS: 70 T1DM patients and 48 healthy volunteers were included. All subjects accepted nerve conduction velocity and CPT examinations for four limbs. Detailed clinical indicators were recorded. CPT values were compared between TIDM group and control group. The risk factors affecting DPN were further explored. RESULTS: Compared with the control group, CPT values under three frequencies were decreased in T1DM group. The abnormality rate of sural nerves was higher than that of median nerves (P<0.001). Median nerve dysfunction mainly presented as hypoesthesia under 250Hz and 5Hz current stimulus. And sural nerve dysfunction mainly presented as hyperesthesia under three frequencies. Compared with left median nerve, abnormal rate of right median nerve was higher under 2000Hz current stimulus (P=0.035). However, abnormal rate of left sural nerve was higher than that of right side under 250Hz and 5Hz current stimulus (P=0.001, <0.001). Duration, NDS scores and CPT values of right median nerve under 2000Hz current stimulus were independent risk factors of abnormal nerve conduction velocity. CONCLUSIONS: The study proved that DPN of T1DM are mainly lower limb-injured., amyelinated and thin myelinated nerve fiber-involved. CPT can be combined with traditional nerve conduction velocity examination, which will help the diagnosis of DPN of T1DM earlier and more comprehensively.