RESUMEN
Vasostatin 1 (VS-1) plays an important role in the regulation of various tissue injury and repair processes, but its role in aortic aneurysm remains unclear. The plasmid-like nanoparticles containing the vasostatin-1 gene Pul-PGEA-pCas-sgVs-1 were constructed, and their guarantee, safety, hemolysis, and particle size were analyzed. Eighty-four eight-week-old male ApoE-mice were randomly divided into blank group (without any treatment), model group (Ang II aortic aneurysm model + tail injection of PBS), control group (modeling + tail injection of Pul-PGEA-pCas9), and experimental group (modeling + tail injection of Pul-PGEA-pCas-sgVs-1), with 21 rats in each group. The incidence, mortality, and maximum diameter of abdominal aortic aneurysm (AAA) and the contents of high sensitivity C-reactive protein (HS-CRP), soluble intercellular adhesion molecule-1 (ICAM-1), soluble vascular cell adhesion molecule-1 (VCAM-1), and TNF-a in serum were compared in different groups of mice. The results showed that Pul-PGEA-pCas-sgVs-1 had good biosafety and transfection ability. The maximum diameter of abdominal aorta, incidence of abdominal aortic aneurysm, mortality, and the expression levels of HS-CRP, ICAM-1, VCAM-1, and TNF-a in the experimental group were lower than those in the model group (P< 0.05). These results indicated that the plasmid-like nanoparticles Pul-PGEA-pCas-sgVs-1 can inhibit the development of aorta by down-regulating the expression of inflammatory factors, which played a good protective role on the aorta.
Asunto(s)
Aneurisma de la Aorta Abdominal , Cromogranina A , Regulación de la Expresión Génica , Nanopartículas , Fragmentos de Péptidos , Plásmidos , Animales , Aneurisma de la Aorta Abdominal/genética , Aneurisma de la Aorta Abdominal/metabolismo , Aneurisma de la Aorta Abdominal/prevención & control , Cromogranina A/biosíntesis , Cromogranina A/genética , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Noqueados para ApoE , Nanopartículas/química , Nanopartículas/uso terapéutico , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/genética , Plásmidos/química , Plásmidos/genética , Plásmidos/farmacologíaRESUMEN
The dopaminergic system plays an essential role in maintaining homeostasis between the central nervous system (CNS) and the immune system. Previous studies have associated imbalances in the dopaminergic system to the pathogenesis of multiple sclerosis (MS). Here, we examined the protein levels of dopaminergic receptors (D1R and D2R) in different phases of the experimental autoimmune encephalomyelitis (EAE) model. We also investigated if the treatment with pramipexole (PPX)-a dopamine D2/D3 receptor-preferring agonist-would be able to prevent EAE-induced motor and mood dysfunction, as well as its underlying mechanisms of action. We report that D2R immunocontent is upregulated in the spinal cord of EAE mice 14 days post-induction. Moreover, D1R and D2R immunocontents in lymph nodes and the oxidative damage in the spinal cord and striatum of EAE animals were significantly increased during the chronic phase. Also, during the pre-symptomatic phase, axonal damage in the spinal cord of EAE mice could already be found. Surprisingly, therapeutic treatment with PPX failed to inhibit the progression of EAE. Of note, PPX treatment inhibited EAE-induced depressive-like while failed to inhibit anhedonic-like behaviors. We observed that PPX treatment downregulated IL-1ß levels and increased BNDF content in the spinal cord after EAE induction. Herein, we show that a D2/D3 receptor-preferred agonist mitigated EAE-induced depressive-like behavior, which could serve as a new possibility for further clinical trials on treating depressive symptoms in MS patients. Thus, we infer that D2R participates in the crosstalk between CNS and immune system during autoimmune and neuroinflammatory response induced by EAE, mainly in the acute and chronic phase of the disease.
Asunto(s)
Encefalomielitis Autoinmune Experimental/metabolismo , Receptores de Dopamina D1/fisiología , Receptores de Dopamina D2/fisiología , Anhedonia/efectos de los fármacos , Anhedonia/fisiología , Animales , Axones/patología , Factor Neurotrófico Derivado del Encéfalo/biosíntesis , Factor Neurotrófico Derivado del Encéfalo/genética , Cuerpo Estriado/metabolismo , Depresión/etiología , Depresión/prevención & control , Progresión de la Enfermedad , Evaluación Preclínica de Medicamentos , Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Encefalomielitis Autoinmune Experimental/psicología , Femenino , Interleucina-1beta/biosíntesis , Interleucina-1beta/genética , Ganglios Linfáticos/metabolismo , Ratones , Ratones Endogámicos C57BL , Enfermedades Neuroinflamatorias/tratamiento farmacológico , Enfermedades Neuroinflamatorias/metabolismo , Estrés Oxidativo , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/genética , Pramipexol/farmacología , Pramipexol/uso terapéutico , Receptores de Dopamina D2/agonistas , Receptores de Dopamina D3/agonistas , Método Simple Ciego , Médula Espinal/metabolismo , Médula Espinal/patologíaRESUMEN
Amyloid beta (Aß) 42 peptide accumulated in Alzheimer disease (AD) patients' brain, often colocalized with serine protease inhibitor family A member 3 (SERPINA3). Being a chaperon, SERPINA3 accelerated Aß42 fibrillization. While analyzing chaperon activity of human SERPINA3 polymorphisms, we found SERPINA3-R124C played a role in protecting cells from Aß42 cytotoxicity. SH-SY5Y cells exposed to Aß42 preincubated with wild-type SERPINA3 (SERPINA3-WT) resulted in extended toxicity leading cell death whereas Aß42 with SERPINA3-R124C resulted in less cytotoxicity. Transmission electron microscope and thioflavin T assay revealed that SERPINA3-R124C shortened lifetime of small soluble oligomer and maintained ß-sheet rich protofibril-like aggregates for longer time compared to that of with SERPINA3-WT. Western blot assay confirmed that SERPINA3-R124C converted Aß42 mostly into high molecular aggregates. Here, we demonstrate first time that polymorphic SERPINA3 acts as a benign chaperon by modulating the transition states of Aß42, which may contribute to the reduction of AD risk.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Biopolímeros/metabolismo , Fragmentos de Péptidos/metabolismo , Serpinas/metabolismo , Péptidos beta-Amiloides/biosíntesis , Péptidos beta-Amiloides/química , Benzotiazoles/metabolismo , Western Blotting , Catálisis , Línea Celular Tumoral , Humanos , Microscopía Electrónica de Transmisión , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/química , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Serpinas/químicaRESUMEN
Sodium restriction is often recommended in heart failure (HF) to block symptomatic edema, despite limited evidence for benefit. However, a low-sodium diet (LSD) activates the classical renin-angiotensin-aldosterone system (RAAS), which may adversely affect HF progression and mortality in patients with dilated cardiomyopathy (DCM). We performed a randomized, blinded pre-clinical trial to compare the effects of a normal (human-equivalent) sodium diet and a LSD on HF progression in a normotensive model of DCM in mice that has translational relevance to human HF. The LSD reduced HF progression by suppressing the development of pleural effusions (p < 0.01), blocking pathological increases in systemic extracellular water (p < 0.001) and prolonging median survival (15%, p < 0.01). The LSD activated the classical RAAS by increasing plasma renin activity, angiotensin II and aldosterone levels. However, the LSD also significantly up-elevated the counter-regulatory RAAS by boosting plasma angiotensin converting enzyme 2 (ACE2) and angiotensin (1-7) levels, promoting nitric oxide bioavailability and stimulating 3'-5'-cyclic guanosine monophosphate (cGMP) production. Plasma HF biomarkers associated with poor outcomes, such as B-type natriuretic peptide and neprilysin were decreased by a LSD. Cardiac systolic function, blood pressure and renal function were not affected. Although a LSD activates the classical RAAS system, we conclude that the LSD delayed HF progression and mortality in experimental DCM, in part through protective stimulation of the counter-regulatory RAAS to increase plasma ACE2 and angiotensin (1-7) levels, nitric oxide bioavailability and cGMP production.
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Angiotensina I/biosíntesis , GMP Cíclico/metabolismo , Dieta Hiposódica , Edema/prevención & control , Insuficiencia Cardíaca/complicaciones , Óxido Nítrico/metabolismo , Fragmentos de Péptidos/biosíntesis , Animales , Disponibilidad Biológica , Biomarcadores/sangre , Presión Sanguínea , Cardiomiopatía Dilatada/complicaciones , Cardiomiopatía Dilatada/fisiopatología , Edema/sangre , Insuficiencia Cardíaca/sangre , Insuficiencia Cardíaca/fisiopatología , Riñón/fisiopatología , Masculino , Ratones Endogámicos C57BL , Péptido Natriurético Encefálico/metabolismo , Óxido Nítrico/sangre , Óxido Nítrico Sintasa/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Derrame Pleural , Sistema Renina-Angiotensina , Análisis de Supervivencia , SístoleRESUMEN
Accumulation of amyloid-ß (Aß) in the brain is a central component of pathology in Alzheimer's disease. A growing volume of evidence demonstrates close associations between periodontal pathogens including Porphyromonas gingivalis (P. gingivalis) and Treponema denticola (T. denticola) and AD. However, the effect and mechanisms of T. denticola on accumulation of Aß remain to be unclear. In this study, we demonstrated that T. denticola was able to enter the brain and act directly on nerve cells resulting in intra- and extracellular Aß1-40 and Aß1-42 accumulation in the hippocampus of C57BL/6 mice by selectively activating both ß-secretase and γ-secretase. Furthermore, both KMI1303, an inhibitor of ß-secretase, as well as DAPT, an inhibitor of γ- secretase, were found to be able to inhibit the effect of T. denticola on Aß accumulation in N2a neuronal cells. Overall, it is concluded that T. denticola increases the expression of Aß1-42 and Aß1-40 by its regulation on beta-site amyloid precursor protein cleaving enzyme-1 and presenilin 1.
Asunto(s)
Péptidos beta-Amiloides/biosíntesis , Hipocampo/metabolismo , Boca/microbiología , Fragmentos de Péptidos/biosíntesis , Treponema denticola/patogenicidad , Infecciones por Treponema/metabolismo , Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Secretasas de la Proteína Precursora del Amiloide/biosíntesis , Secretasas de la Proteína Precursora del Amiloide/genética , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Animales , Aorta/microbiología , Ácido Aspártico Endopeptidasas/biosíntesis , Ácido Aspártico Endopeptidasas/genética , Diaminas/farmacología , Activación Enzimática , Hipocampo/microbiología , Masculino , Ratones , Ratones Endogámicos C57BL , Neuronas/metabolismo , Neuronas/microbiología , Porphyromonas gingivalis/patogenicidad , Presenilina-1/biosíntesis , Presenilina-1/genética , Distribución Aleatoria , Tiazoles/farmacología , Infecciones por Treponema/patología , Ganglio del Trigémino/metabolismo , Ganglio del Trigémino/microbiologíaRESUMEN
INTRODUCTION: A variety of transgenic and knock-in mice that express mutant alleles of Amyloid precursor protein (APP) have been used to model the effects of amyloid-beta (Aß) on circuit function in Alzheimer's disease (AD); however phenotypes described in these mice may be affected by expression of mutant APP or proteolytic cleavage products independent of Aß. In addition, the effects of mutant APP expression are attributed to elevated expression of the amyloidogenic, 42-amino acid-long species of Aß (Aß42) associated with amyloid plaque accumulation in AD, though elevated concentrations of Aß40, an Aß species produced with normal synaptic activity, may also affect neural function. METHODS: To explore the effects of elevated expression of Aß on synaptic function in vivo, we assessed visual system plasticity in transgenic mice that express and secrete Aß throughout the brain in the absence of APP overexpression. Transgenic mice that express either Aß40 or Aß42 were assayed for their ability to appropriately demonstrate ocular dominance plasticity following monocular deprivation. RESULTS: Using two complementary approaches to measure the plastic response to monocular deprivation, we find that male and female mice that express either 40- or 42-amino acid-long Aß species demonstrate a plasticity defect comparable to that elicited in transgenic mice that express mutant alleles of APP and Presenilin 1 (APP/PS1 mice). CONCLUSIONS: These data support the hypothesis that mutant APP-driven plasticity impairment in mouse models of AD is mediated by production and accumulation of Aß. Moreover, these findings suggest that soluble species of Aß are capable of modulating synaptic plasticity, likely independent of any aggregation. These findings may have implications for the role of soluble species of Aß in both development and disease settings.
Asunto(s)
Péptidos beta-Amiloides/biosíntesis , Predominio Ocular/fisiología , Plasticidad Neuronal/fisiología , Fragmentos de Péptidos/biosíntesis , Corteza Visual/metabolismo , Péptidos beta-Amiloides/genética , Animales , Femenino , Masculino , Ratones , Ratones Transgénicos , Fragmentos de Péptidos/genéticaRESUMEN
BACKGROUND AND AIMS: Levosimendan, a calcium (Ca2+)-sensitizing cardiotonic agent, is mainly used in patients with advanced heart failure. However, no research could explain how levosimendan reduces the mortality in advanced heart failure patients. We aim to illustrate the efficacy of levosimendan through clinical indexes. METHODS: We searched PubMed, Embase, and CENTRAL from 1994 to August 2019 to compare the efficacy of levosimendan infusion for the treatment of advanced heart failure with that of other agents (placebo, dobutamine, furosemide, and prostaglandin E1). Levels of B-type natriuretic peptide (BNP) and N-terminal pro BNP (NT-proBNP), and left ventricular ejection fraction (LVEF) and heart rate (HR) were analyzed. The count data were analyzed by the standardized mean difference (SMD) and its 95% confidence interval (CI) to determine the effect size. We chose the random effect model or the fixed effect model according to the heterogeneity. RESULTS: Nine randomized controlled trials with 413 patients were ultimately enrolled. Compared with other agents (placebo, dobutamine, furosemide, and prostaglandin E1), levosimendan significantly reduced the BNP level (SMD - 0.91; 95% CI - 1.44 to - 0.39; p = 0.001; I2 = 74.3%) and improved the LVEF (SMD 0.74; 95% CI 0.22-1.25; p = 0.005; I2 = 79.7%). However, levosimendan did not significantly change the HR (SMD 0.09; 95% CI - 0.24 to 0.42; p = 0.592; I2 = 51.5%). Meanwhile, we found that the main source of heterogeneity was the use of loaded or unloaded levosimendan. CONCLUSION: Our meta-analysis suggests that intravenous levosimendan can reduce BNP level and increase LVEF in patients with advanced heart failure to reduce the mortality at the shortest follow-up available.
Asunto(s)
Cardiotónicos/farmacología , Insuficiencia Cardíaca/tratamiento farmacológico , Péptido Natriurético Encefálico/biosíntesis , Simendán/farmacología , Volumen Sistólico/efectos de los fármacos , Cardiotónicos/uso terapéutico , Hemodinámica/efectos de los fármacos , Humanos , Péptido Natriurético Encefálico/sangre , Fragmentos de Péptidos/biosíntesis , Ensayos Clínicos Controlados Aleatorios como Asunto , Simendán/uso terapéutico , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
Imidazolium based IL's has gained vast interest in developing biological applications. Oligomerization and fibrillization of amyloid ß (1-42) peptide are mainly responsible for the extra-neuronal deposition of amyloid fibrils in neurodegenerative disorders like Alzheimer's disease (AD). Here, we report an effect of tert-BuOH-functional imidazolium ILs on oligomerization and fibrillization of amyloid ß (1-42) Peptide in vitro. In this study, a series of these [alkyl-tOHim][OMs] ILs with methyl sulphonate counter anion by varying alkyl chains were used. Among the seven protic ILs, four showed strong binding and inhibition activity for the formation of amyloid ß (1-42) aggregation by using Thioflavin T fluorescence binding assay. The secondary structural analysis of the peptide, pre-incubated with active ILs shows the loss of ordered ß-sheet amyloid structure. The longer alkyl chain ILs showed that an increased in amyloid binding and hence an inhibition effect on amyloid aggregation was enhanced. Thus, we propose that ILs could be presented as potential candidates for therapeutic intervention against Alzheimer's disease (AD).
Asunto(s)
Péptidos beta-Amiloides/antagonistas & inhibidores , Imidazoles/farmacología , Líquidos Iónicos/farmacología , Fragmentos de Péptidos/antagonistas & inhibidores , Agregado de Proteínas/efectos de los fármacos , Alcohol terc-Butílico/farmacología , Péptidos beta-Amiloides/biosíntesis , Imidazoles/síntesis química , Imidazoles/química , Líquidos Iónicos/síntesis química , Líquidos Iónicos/química , Microscopía Electrónica de Transmisión , Fragmentos de Péptidos/biosíntesis , Sales (Química)/síntesis química , Sales (Química)/química , Sales (Química)/farmacología , Alcohol terc-Butílico/químicaRESUMEN
Previously, we revealed that brain Ang-(1-7) deficiency was involved in the pathogenesis of sporadic Alzheimer's disease (AD). We speculated that restoration of brain Ang-(1-7) levels might have a therapeutic effect against AD. However, the relatively short duration of biological effect limited the application of Ang-(1-7) in animal experiments. Since Ang-(1-7) is generated by its metabolic enzyme ACE2, we then tested the efficacy of an ACE2 activator diminazene aceturate (DIZE) on AD-like neuropathology and cognitive impairment in senescence-accelerated mouse prone substrain 8 (SAMP8) mice, an animal model of sporadic AD. Eight-month-old SAMP8 mice were injected intraperitoneally with vehicle or DIZE once a day for 30 consecutive days. DIZE markedly elevated brain Ang-(1-7) and MAS1 levels. Meanwhile, DIZE significantly reduced the levels of Aß1-42, hyperphosphorylated tau and pro-inflammatory cytokines in the brain. The synaptic and neuronal losses in the brain were ameliorated by DIZE. Importantly, DIZE improved spatial cognitive functions in the Morris water maze test. In conclusion, this study demonstrates that DIZE ameliorates AD-like neuropathology and rescues cognitive impairment in SAMP8 mice. These beneficial effects of DIZE may be achieved by activating brain ACE2/Ang-(1-7)/MAS1 axis. These findings highlight brain ACE2/Ang-(1-7)/MAS1 axis as a potential target for the treatment of sporadic AD.
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Enfermedad de Alzheimer/tratamiento farmacológico , Enzima Convertidora de Angiotensina 2/efectos de los fármacos , Disfunción Cognitiva/tratamiento farmacológico , Diminazeno/análogos & derivados , Enfermedad de Alzheimer/patología , Enfermedad de Alzheimer/psicología , Péptidos beta-Amiloides/biosíntesis , Péptidos beta-Amiloides/genética , Angiotensina I/metabolismo , Animales , Química Encefálica/efectos de los fármacos , Química Encefálica/genética , Disfunción Cognitiva/etiología , Citocinas/biosíntesis , Diminazeno/uso terapéutico , Infusiones Parenterales , Masculino , Aprendizaje por Laberinto , Ratones , Ratones Mutantes Neurológicos , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas/genética , Receptores Acoplados a Proteínas G/genética , Proteínas tau/biosíntesisRESUMEN
Objective It is well known that poor sleep increases the risk of heart failure (HF). However, the underlying mechanisms remain unclear. In this study, we investigated the association of poor sleep with hemodynamic stress on the left ventricle, which was a key factor for the development of HF in elderly individuals. Methods A total of 2,301 participants (≥65 years old) without cardiac disease were enrolled in this cross-sectional analysis. We evaluated the subjective sleep quality, sleeping difficulty, subjective sleep duration, use of sleeping pills, and daytime dysfunction using the Pittsburgh Sleep Quality Index, a 19-item self-reported questionnaire. We assessed serum N-terminal pro-brain natriuretic peptide (NT-proBNP) as a marker of hemodynamic stress on the left ventricle, and we defined high NT-proBNP as a serum NT-proBNP level ≥ 125 pg/mL. Results Sleeping difficulty was significantly associated with high NT-proBNP levels [odds ratio (OR), 1.46; 95% confidence interval (CI), 1.16-1.85; p<0.005]. A subjective short sleep duration was also significantly associated with high NT-proBNP levels (OR, 1.69; 95% CI, 1.03-2.75; p<0.05). A subjective poor sleep quality, the use of sleeping pills, and daytime dysfunction were not associated with serum NT-proBNP levels. All data were adjusted for the age, sex, body mass index, serum hemoglobin concentration, serum creatinine level, systolic blood pressure, diastolic blood pressure, and use of antihypertensive medications. Conclusion Poor sleep was associated with high hemodynamic stress to the left ventricle in elderly population.
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Hemodinámica/fisiología , Péptido Natriurético Encefálico/biosíntesis , Fragmentos de Péptidos/biosíntesis , Trastornos del Sueño-Vigilia/fisiopatología , Sueño/fisiología , Factores de Edad , Anciano , Anciano de 80 o más Años , Biomarcadores , Presión Sanguínea/fisiología , Índice de Masa Corporal , Comorbilidad , Estudios Transversales , Femenino , Ventrículos Cardíacos/fisiopatología , Humanos , Hipnóticos y Sedantes/farmacología , Masculino , Oportunidad Relativa , Factores SexualesRESUMEN
BACKGROUND: Tetanus is an infectious disease caused by Clostridium secreting tetanus toxin in anaerobic environment. The fragment C of Tetanus toxin (TTc) has been widely studied as a candidate vaccine to replace the existing tetanus toxoid vaccine. OBJECTIVE: In this study, we established a simple method to purify recombinant protein TTc with ion-exchange chromatography from Escherichia coli expression systems. METHODS: The TTc gene sequence was cloned into pET26b (+) vector and transferred to E. coli BL21 (DE3) for expression. The fermentation conditions (IPTG concentration, Induction temperature, Induction time) were optimized to obtain more soluble proteins. The soluble proteins were purified by Anion exchange chromatography and Cation exchange chromatography. The sequence of columns in the purification process was discussed. Finally, the stability of purified TTc protein were determined, the secondary structure of the purified TTc protein was determined by circular dichroism. The molecular weight of the purified TTc protein was determined by liquid chromatograph- mass spectrometer. Furthermore, we verified the immunogenicity of the purified protein in mice. RESULTS: The purity of TTc improved from 34% to 88% after the first anion exchange column, and the final yield of recombinant TTc (purity > 95%) can reach 84.79% after the following cation exchange chromatography. The recombinant TTc had a molecular weight of 51.737 KDa, was stable at 4 °C and weak alkaline environment, was a ß-sheet secondary structure, and had strong immunogenicity. CONCLUSION: The purification method we developed might be an efficient method for the industrial production of tetanus recombinant TTc vaccine.
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Expresión Génica , Fragmentos de Péptidos , Toxina Tetánica , Escherichia coli/química , Escherichia coli/genética , Escherichia coli/metabolismo , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/aislamiento & purificación , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Toxina Tetánica/biosíntesis , Toxina Tetánica/química , Toxina Tetánica/genética , Toxina Tetánica/aislamiento & purificaciónRESUMEN
BACKGROUND: Dysfunction of synaptic plasticity leads to memory impairment in Alzheimer's disease (AD). Muscone (Mus) has shown neuroprotective effects in cerebral ischemic models. However, little is known of Mus effects on AD. OBJECTIVE: To investigate the effects of Mus on memory functions and synaptic plasticity in 6-month-old APP/PS1 double-transgenic mice and explore the potential mechanisms. METHODS: Mus was intraperitoneally injected into APP/PS1 or wild-type mice, and cognitive function was assessed by Novel object recognition and Morris water maze tests. The levels of amyloid-ß (Aß) were evaluated by immunofluorescence staining and ELISA. Synaptic morphology and plasticity were evaluated by Golgi staining and long-term potentiation. Cell viability was examined by Cell Counting Kit-8 assay. The protein levels of histone deacetylase 2 (HDAC2) were accessed by western blotting and Immunofluorescence staining. The protein levels of microtubule associated protein 2 and synaptophysin were analyzed by immunofluorescence staining. The ubiquitination of HDAC2 was examined by co-immunoprecipitation. The interaction of Mus with HDAC2 was predicted by molecular docking analysis. RESULTS: Mus treatment attenuated memory dysfunction, reduced Aß level, and enhanced synaptic plasticity in APP/PS1 mice. In addition, Mus treatment decreased the level of HDAC2 in the hippocampus of APP/PS1 mice and Aß1-42-induced primary neurons, which might be associated with increased HDAC2 ubiquitination induced by HDAC2 and Mus interaction. CONCLUSION: Mus protected against synaptic plasticity and memory impairment in APP/PS1 mice, and enhanced HDAC2 degradation via ubiquitination, indicating that Mus was a potential drug for AD treatment.
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Precursor de Proteína beta-Amiloide/genética , Disfunción Cognitiva/tratamiento farmacológico , Disfunción Cognitiva/genética , Cicloparafinas/uso terapéutico , Plasticidad Neuronal/efectos de los fármacos , Presenilina-1/genética , Péptidos beta-Amiloides/antagonistas & inhibidores , Péptidos beta-Amiloides/biosíntesis , Péptidos beta-Amiloides/química , Animales , Células Cultivadas , Disfunción Cognitiva/patología , Cicloparafinas/farmacología , Hipocampo/efectos de los fármacos , Hipocampo/patología , Hipocampo/fisiología , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Plasticidad Neuronal/fisiología , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/química , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Sinapsis/efectos de los fármacos , Sinapsis/patología , Sinapsis/fisiologíaRESUMEN
OBJECTIVE: The obtain purified recombinant asprosin and test its functions. METHODS: The recombinant plasmid of pET-22b-asprosin was constructed and transformed into competent E.coli BL (DE3) strain. After IPTG-induced expression, asprosin inclusion body was renatured by gradient urea and purified by Ni-NTA affinity chromatography column followed by removal of endotoxin to obtain recombinant asprosin for use in cells and animals experiments. C57 mice were injected intraperitoneally with the recombinant asprosin and blood glucose was detected using a blood glucose meter. Alamar Blue assay was used to evaluate of the effect of the recombinant asprosin on the viability of MIHA cells, and cellular glycogen content was detected using the anthrone method. RESULTS: At the absorbance at 600 nm of 0.8, induction of the recombinant host bacteria with 1 mmol/L IPTG at 37 â for 4 h optimally induced the expression of asprosin inclusion body. After purification and endotoxin removal, the purity of the recombinant asprosin exceeded 95% with the content of endotoxin below 1 EU/mg. In C57 mice, intraperitoneal injection with recombinant asprosin significantly increased blood glucose level, which reached the peak level at 60 min following the injection (P=0.021) and recovered the normal level at 120 min (P=0.03). Treatment with the recombinant asprosin for 24 h did not cause obvious adverse effect on the viability of MIHA cells but significantly lowered glycogen content in the cells (P < 0.05). CONCLUSIONS: We successfully obtained recombinant asprosin using a prokaryotic expression system. The recombinant asprosin can decrease glycogen content in MIHA cells and increase blood glucose level in mice.
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Cuerpos de Inclusión , Proteínas de Microfilamentos/biosíntesis , Fragmentos de Péptidos/biosíntesis , Hormonas Peptídicas/biosíntesis , Animales , Glucemia/análisis , Línea Celular , Escherichia coli , Fibrilina-1 , Glucógeno/análisis , Humanos , Ratones , Ratones Endogámicos C57BL , Plásmidos , Proteínas Recombinantes/biosíntesisRESUMEN
Alzheimer's disease (AD) is one of the most common neurodegenerative diseases, and no effective therapies have been found to prevent or cure AD to date. Berberine and curcumin are extracts from traditional Chinese herbs that have a long history of clinical benefits for AD. Here, using a transgenic AD mouse model, we found that the combined berberine and curcumin treatment had a much better effect on improving the cognitive function of mice than the single-drug treatment, suggesting synergic effects of the combined berberine and curcumin treatment. In addition, we found that the combined berberine and curcumin treatment had significant synergic effects on reducing soluble amyloid-ß-peptide(1-42) production. Furthermore, the combination treatment also had remarkable synergic effects on decreasing inflammatory responses and oxidative stress in both the cortex and hippocampus of AD mice. We also found that the combination treatment performed much better than the single drugs in reducing the APP and BACE1 levels and increasing AMPKα phosphorylation and cell autophagy, which might be the underlying mechanism of the synergic effects. Taken together, the result of this study reveal the synergic effects and potential underlying mechanisms of the combined berberine and curcumin treatment in improving the symptoms of AD in mice. This study sheds light on a new strategy for exploring new phytotherapies for AD and also emphasizes that more research should focus on the synergic effects of herbal drugs in the future.
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Enfermedad de Alzheimer/tratamiento farmacológico , Berberina/administración & dosificación , Encéfalo/efectos de los fármacos , Cognición/efectos de los fármacos , Curcumina/administración & dosificación , Estrés Oxidativo/efectos de los fármacos , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Secretasas de la Proteína Precursora del Amiloide/biosíntesis , Péptidos beta-Amiloides/antagonistas & inhibidores , Péptidos beta-Amiloides/biosíntesis , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Ácido Aspártico Endopeptidasas/antagonistas & inhibidores , Ácido Aspártico Endopeptidasas/biosíntesis , Encéfalo/metabolismo , Cognición/fisiología , Sinergismo Farmacológico , Femenino , Masculino , Ratones , Ratones Transgénicos , Estrés Oxidativo/fisiología , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/biosíntesisRESUMEN
Proteins and peptides account for 20-75% of marine biota biomass, of which a major fraction is metabolized by bacteria, thus deciphering interactions between bacteria and peptides is important in understanding marine carbon and nitrogen cycling. To better understand capabilities of different bacterial strains on peptide decomposition, four Gammaproteobacteria (Pseudoalteromonas atlantica, Alteromonas sp., Marinobacterium jannaschii, Amphritea japonica) were incubated in autoclaved seawater amended with tetrapeptide alanine-valine-phenylalanine-alanine (AVFA), a fragment of RuBisCO. While AVFA was decomposed greatly by Pseudoalteromonas atlantica and Alteromonas sp, it remained nearly intact in the Marinobacterium jannaschii and Amphritea japonica incubations. Pseudoalteromonas and Alteromonas decomposed AVFA mainly through extracellular hydrolysis pathway, releasing 71-85% of the AVFA as hydrolysis products to the surrounding seawater. Overall, this study showed that Gammaproteobacterial strains differ greatly in their capabilities of metabolizing peptides physiologically, providing insights into interactions of bacteria and labile organic matter in marine environments.
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Gammaproteobacteria/metabolismo , Fragmentos de Péptidos/metabolismo , Agua de Mar/química , Aminoácidos/biosíntesis , Hidrólisis , Fragmentos de Péptidos/biosíntesis , Agua de Mar/microbiologíaRESUMEN
OBJECTIVES: Thyroid hormones have important roles in normal development and energy regulating mechanisms as well as signaling mechanisms that affect energy consumption through central and peripheral pathways. The aim of this study was to determine the effects of thyroid dysfunction on adropin, asprosin and preptin levels in rat. METHODS: The study was performed on the 38 male Wistar-albino rats. Experiment groups were designed as follows. 1-Control, 2-Hypothyroidism; To induce hypothyroidism PTU was applied by intraperitoneal as 10 mg/kg/day for 2 weeks. 3-Hypothyroidism + Thyroxine; Previously animals were made with hypothyroidism by 1 week PTU application and then 1 week l-thyroxine was given by intraperitoneal as 1.5 mg/kg/day. 4-Hyperthyroidism; Rats were made with hyperthyroidism by 3 weeks l-thyroxine (0.3 mg/kg/day). 5-Hyperthyroidism + PTU; Animals were made hyperthyroisim by l-thyroxine as groups 4, then 1 week PTU was applied to treatment of hiperthyrodism. At the end of supplementation animals were sacrificed and blood samples were collected for FT3, FT4, adropin, asprosin, preptin analysis. RESULTS: FT3 ve FT4 levels were reduced significantly in hypothyroidism while increased in hyperthyroidism (p<0.001). Hipothyrodism led to reduces adropin, asprosin and preptin levels. And also hyperthyroidism reduced adropin and preptin levels (p<0.001). CONCLUSIONS: The results of study show that experimental hypothyroidism and hyperthyroidism lead to significantly change to adropin, asprosin and preptin levels. However, correction of thyroid function caused to normals levels in asprosin and preptin.
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Fibrilina-1/sangre , Hipertiroidismo/metabolismo , Hipotiroidismo/metabolismo , Fragmentos de Péptidos/sangre , Hormonas Peptídicas/sangre , Péptidos/sangre , Tiroxina/sangre , Triyodotironina/sangre , Animales , Proteínas Sanguíneas/biosíntesis , Fibrilina-1/biosíntesis , Hipertiroidismo/inducido químicamente , Hipotiroidismo/inducido químicamente , Factor II del Crecimiento Similar a la Insulina/biosíntesis , Fragmentos de Péptidos/biosíntesis , Hormonas Peptídicas/biosíntesis , Propiltiouracilo/toxicidad , Ratas , Tiroxina/biosíntesis , Tiroxina/toxicidad , Triyodotironina/biosíntesisRESUMEN
OBJECTIVES: The activation of the adenosine A3 receptor (A3AR) can regulate inflammation, but the way that this regulates colonic mucosal inflammation in ulcerative colitis (UC) remains unclear. This study aimed at examining A3AR expression and investigating the effect of A3AR activation on ex vivo cytokine expression and nuclear factor-kappa B (NF-κB) signaling in colonic mucosa. METHODS: Colonic mucosal biopsied tissue from 18 patients with UC and 11 healthy controls was tested for A3AR expression by immunofluorescence, quantitative real-time polymerase chain reaction and Western blot. Following treatment for 24 hours with or without 2-Cl-IB-MECA, an A3AR agonist, TNF-α and IL-1ß secreted by the cultured colonic mucosal tissue were quantified by ELISA. The colonic mucosal epithelia were dissected and treated with, or without 2-Cl-IB-MECA for 24 hours. The NF-κB p65 protein and its distribution in the cultured colonic epithelia were examined by immunofluorescence and Western blot. RESULTS: Compared with the controls, down-regulated A3AR expression and up-regulated TNF-α and IL-1ß production and NF-κB p65 protein were observed in the UC colonic mucosa. The activation of A3AR by 2-Cl-IB-MECA significantly decreased TNF-α and IL-1ß production and attenuated the NF-κB p65 activation in colonic tissues from patients with UC. CONCLUSIONS: A3AR activation inhibited inflammation by mitigating pro-inflammatory cytokine production and the NF-κB signal activation in colonic mucosa of patients with UC. A3AR activation may play a role in the pathogenesis of UC.
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Adenosina/análogos & derivados , Colitis Ulcerosa/inmunología , Colon/inmunología , FN-kappa B/metabolismo , Agonistas del Receptor Purinérgico P1/farmacología , Receptor de Adenosina A3/inmunología , Adenosina/farmacología , Adenosina/uso terapéutico , Colitis Ulcerosa/tratamiento farmacológico , Colon/efectos de los fármacos , Citocinas/antagonistas & inhibidores , Citocinas/biosíntesis , Citocinas/inmunología , Regulación hacia Abajo , Humanos , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Interleucina-1beta/biosíntesis , Interleucina-1beta/inmunología , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/inmunología , FN-kappa B/biosíntesis , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/inmunología , Agonistas del Receptor Purinérgico P1/uso terapéutico , Receptor de Adenosina A3/biosíntesis , Transducción de Señal , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/inmunología , Regulación hacia ArribaRESUMEN
INTRODUCTION: The 2011 WHO Classification for lung adenocarcinoma enlightened the need for a wise use of immunohistochemistry to preserve tissue for both diagnosis and molecular studies. The current recommendation is to use a panel comprising TTF1 and p40 to classify tumors with no clear squamous or glandular differentiation as many studies have showed the higher specificity of p40 over p63 as marker of squamous differentiation. However, the co-expression of both markers opens a new scenario with subsequent classification and potentially treatment issues. MATERIALS AND METHODS: We report a case of a non-small lung cell carcinoma (NSCLC) with coexistent expression of TTF1 and p40 in the same tumour cells. To our knowledge, this peculiar immunohistochemical profile is very rare, and thus a review of the clinical and molecular features including molecular variances of the tumour was performed. Review of the pertinent literature was also carried out. RESULTS: Two additional articles describing unusual cases of NSCLC with coexistent expression of TTF1 and p40 were found and compared to our case. Interestingly, they all carried out aberrant mutation in TP53 oncogene and were of advance stage. CONCLUSION: The positivity for both "squamous" and "adenocarcinomatous" markers and mutations of TP53 could be the expression of a not fully recognized variant of NSCLC with possible implications for classification, diagnosis and therapy.
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Carcinoma de Pulmón de Células no Pequeñas/genética , Proteínas de Unión al ADN/genética , Regulación Neoplásica de la Expresión Génica , Epítopos Inmunodominantes/genética , Neoplasias Pulmonares/genética , Mutación/genética , Fragmentos de Péptidos/genética , Factores de Transcripción/genética , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Proteínas de Unión al ADN/biosíntesis , Humanos , Epítopos Inmunodominantes/biosíntesis , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/metabolismo , Masculino , Persona de Mediana Edad , Fragmentos de Péptidos/biosíntesis , Factores de Transcripción/biosíntesisRESUMEN
OBJECTIVE: We evaluated the ability of Coll2-1, a type II collagen peptide, to activate pro-inflammatory pathways in synovial cells and to induce arthritis in Lewis rats. METHOD: Human synoviocytes and chondrocytes from knee OA patients were cultured for 24 h with/without Coll2-1 and/or purified immunoglobulin G (AS0619) binding specifically this peptide, and/or CLI-095, a TLR-4 signaling inhibitor and/or apocynin and diphenyleneiodonium, Reactive oxygen species (ROS) production inhibitors. The Interleukin (IL)-8 and Vascular Endothelium Growth Factor (VEGF) expression, the IL-8 production, the IκB-α and p65 phosphorylation and ROS were evaluated. Coll2-1 peptide, bovine type II collagen (CIA), streptococcal cell wall (SCW) or saline solution were injected into Lewis rats. The Coll2-1 peptide was injected subcutaneously (SC; 20-200µg/100µl/animal) or intra-articularly (IA; 0.5-5µg/50µl/animal) and compared to CIA injected in SC (200µg/100µl/animal) and SCW in IA (5µg/50µl/animal). The animals were injected on day 0 and monitored for 28 days. Histological lesions assessment was performed using an arthritis score. RESULTS: Coll2-1 peptide significantly increased IL-8 gene expression and production by synoviocytes. AS0619 and CLI-095 significantly decreased IL-8 expression. Coll2-1 induced p65 and IκBα phosphorylation and oxidative stress inhibitors decreased it. In human chondrocytes culture, Coll2-1 significantly increased MMP-3 and VEGF gene expression. In Lewis rats, CIA, SCW or Coll2-1 injection triggered arthritis. Like CIA or SCW, Coll2-1 induced synovitis, loss of cartilage proteoglycans, cartilage structure lesion and subchondral bone remodeling. CONCLUSIONS: Coll2-1 activates synoviocytes to produce IL-8 and induces arthritis in rat. These findings suggest that neutralizing Coll2-1 could be a therapeutic approach of arthritis.
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Colágeno Tipo II/genética , Regulación de la Expresión Génica , Estrés Oxidativo , Fragmentos de Péptidos/genética , ARN/genética , Sinoviocitos/metabolismo , Sinovitis/genética , Anciano , Animales , Células Cultivadas , Colágeno Tipo II/biosíntesis , Modelos Animales de Enfermedad , Femenino , Humanos , Interleucina-8/biosíntesis , Interleucina-8/genética , Masculino , Persona de Mediana Edad , Fragmentos de Péptidos/biosíntesis , Ratas , Ratas Endogámicas Lew , Sinoviocitos/patología , Sinovitis/metabolismo , Sinovitis/patologíaRESUMEN
BACKGROUND: Migraine is a neurovascular primary headache disorder, which causes significant socioeconomic problems worldwide. The pathomechanism of disease is enigmatic, but activation of the trigeminovascular system (TS) appears to be essential during the attack. Migraine research of recent years has focused on neuropeptides, such as calcitonin gene-related peptide (CGRP) and pituitary adenylate cyclase-activating polypeptide 1-38 (PACAP1-38) as potential pathogenic factors and possible therapeutic offensives. The goal of present study was to investigate the simultaneous expression of CGRP and precursor of PACAP1-38 (preproPACAP) in the central region of the TS in a time-dependent manner following TS activation in rats. METHODS: The right whisker pad of rats was injected with 50 µl Complete Freund's Adjuvant (CFA) or saline. A mechanical allodynia test was performed with von Frey filaments before and after treatment. Transcardial perfusion of the animals was initiated 24, 48, 72 and 120 h after injection, followed by the dissection of the nucleus trigeminus caudalis (TNC). After preparation, the samples were stored at - 80 °C until further use. The relative optical density of CGRP and preproPACAP was analyzed by Western blot. One-way ANOVA and Kruskal-Wallis followed by Tukey post hoc test were used to evaluate the data. Regression analysis was applied to explore the correlation between neuropeptides expression and hyperalgesia. RESULTS: Orofacial CFA injection resulted in significant CGRP and preproPACAP release in the TNC 24, 48, 72 and 120 h after the treatment. The level of neuropeptides reached its maximum at 72 h after CFA injection, corresponding to the peak of facial allodynia. Negative, linear correlation was detected between the expression level of neuropeptides and value of mechanonociceptive threshold. CONCLUSION: This is the first study which suggests that the expression of CGRP and preproPACAP simultaneously increases in the central region of activated TS and it influences the formation of mechanical hyperalgesia. Our results contribute to a better understanding of migraine pathogenesis and thereby to the development of more effective therapeutic approaches.