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Engrailed-1 (EN1) is a developmental gene that encodes En1, a highly conserved transcription factor involved in regionalization during early embryogenesis and in the later maintenance of normal neurons. After birth, EN1 still plays a role in the development and physiology of the body; for example, it exerts a protective effect on midbrain dopaminergic (mDA) neurons, and loss of EN1 causes mDA neurons in the ventral midbrain to gradually die approximately 6 weeks after birth, resulting in motor and nonmotor symptoms similar to those observed in Parkinson's disease. Notably, EN1 has been identified as a possible susceptibility gene for idiopathic Parkinson's disease in humans. EN1 is involved in the processes of wound-healing scar production and tissue and organ fibrosis. Additionally, EN1 can lead to tumorigenesis and thus provides a target for the treatment of some tumors. In this review, we summarize the effects of EN1 on embryonic organ development, describe the consequences of the deletion or overexpression of the EN1 gene, and discuss the pathways in which EN1 is involved. We hope to clarify the role of EN1 as a developmental gene and present potential therapeutic targets for diseases involving the EN1 gene.
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Proteínas de Homeodomínio , Doença de Parkinson , Humanos , Proteínas de Homeodomínio/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Neurônios/metabolismo , Regulação da Expressão Gênica , Genes Homeobox , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , Doença de Parkinson/patologiaRESUMO
The double-stranded DNA (dsDNA) viruses represented by adenovirus and monkeypox virus, have attracted widespread attention due to their high infectivity. In 2022, the global outbreak of mpox (or monkeypox) has led to the declaration of a Public Health Emergency of International Concern. However, to date therapeutics approved for dsDNA virus infections remain limited and there are still no available treatments for some of these diseases. The development of new therapies for treating dsDNA infection is in urgent need. In this study, we designed and synthesized a series of novel disulfide-incorporated lipid conjugates of cidofovir (CDV) as potential candidates against dsDNA viruses including vaccinia virus (VACV) and adenovirus (AdV) 5. The structure-activity relationship analyses revealed that the optimum linker moiety was C2H4 and the optimum aliphatic chain length was 18 or 20 atoms. Among the synthesized conjugates, 1c exhibited more potency against VACV (IC50 = 0.0960 µM in Vero cells; IC50 = 0.0790 µM in A549 cells) and AdV5 (IC50 = 0.1572 µM in A549 cells) than brincidofovir (BCV). The transmission electron microscopy (TEM) images revealed that the conjugates could form micelles in phosphate buffer. The stability studies in the GSH environment demonstrated that the formation of micelles in phosphate buffer might protect the disulfide bond from glutathione (GSH) reduction. The dominant means of the synthetic conjugates to liberate the parent drug CDV was by enzymatic hydrolysis. Furthermore, the synthetic conjugates remained sufficiently stable in simulated gastric fluid (SGF), simulated intestinal fluid (SIF), and pooled human plasma, which indicated the possibility for oral administration. These results indicated 1c may be a broad-spectrum antiviral candidate against dsDNA viruses with potential oral administration. Moreover, modification of the aliphatic chain attached to the nucleoside phosphonate group was involved as an efficient prodrug strategy for the development of potent antiviral candidates.
Assuntos
Antivirais , Pró-Fármacos , Animais , Chlorocebus aethiops , Humanos , Cidofovir/farmacologia , Antivirais/química , Pró-Fármacos/farmacologia , Células Vero , Micelas , Citosina/farmacologia , Citosina/química , Vaccinia virus , Lipídeos , FosfatosRESUMO
INTRODUCTION: The purpose of this study was to evaluate the long-term outcomes of posterior capsular opacification (PCO) in highly myopic eyes and its influencing factors. METHODS: Patients undergoing phacoemulsification with intraocular lens implantation and followed up for 1-5 years were included in this prospective cohort study. The severity of PCO was evaluated using EPCO2000 software system, with the area of central 3.0 mm (PCO-3 mm) and within the capsulorhexis (PCO-C) both being analyzed. Percentage of eyes after Nd:YAG capsulotomy, as well as clinically significant PCO (defined as eyes with visual-impairing PCO or after capsulotomy), were also included as outcome variables. RESULTS: A total of 673 highly myopic eyes [axial length (AL) ≥ 26 mm] and 224 control eyes (AL < 26 mm) were analyzed. The mean follow-up time was 34.0 ± 9.0 months. PCO was more severe in highly myopic eyes compared with controls with regard to higher EPCO scores (P < 0.001 for both PCO-3 mm and PCO-C), higher capsulotomy rate (P = 0.001), higher clinically significant PCO rate (P < 0.001) and shorter PCO-free survival time (P < 0.001). Extreme myopia (AL ≥ 28 mm) would further aggravate PCO in terms of higher EPCO scores (PCO-3 mm: P = 0.017; PCO-C: P = 0.013) and higher clinically significant PCO rate (P = 0.024) compared with other myopic eyes. In highly myopic eyes, AL [odds ratio (OR) 1.124, P = 0.004] and follow-up duration (OR 1.082, P < 0.001) were independent risk factors for clinically significant PCO after cataract surgery. CONCLUSION: Highly myopic eyes had more severe PCO in the long term. Longer AL and follow-up duration were associated with higher risk of PCO. CLINICAL TRIAL REGISTRATION: The study was registered at ClinicalTrials.gov (NCT03062085).
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Despite considerable advances in prevention, diagnosis, and therapy, nephrotic syndrome (NS) remains a significant cause of high morbidity and mortality globally. As a result, there is an urgent need to identify novel effective preventative and therapeutic agents for NS. NS is implicated in glomerular permselectivity injury, which can be attributed to oxidative distress, inflammation, lipid nephrotoxicity, podocyte apoptosis, autophagy dysfunction, and slit diaphragm (SLD) dysfunction. In addition to its well-documented antioxidant potency, procyanidin B2 (PB2) may exhibit pleiotropic effects by targeting various canonical signaling events, such as NF-κB, PPARs, PI3K/Akt, mTOR, and the caspase family. As a result, PB2 may be a promising therapeutic target against NS. To test this hypothesis, we established an Adriamycin (ADR)-induced NS mouse model to evaluate the pleiotropic renoprotective effects of PB2 on NS. Here, we demonstrated that PB2 improves podocyte injury via inhibition of NOX4/ROS and Hsp90/NF-κB to exhibit antioxidant and anti-inflammatory potency, respectively. We also show that PB2 indirectly activates the PI3K/Akt axis by regulating SLD protein levels, resulting in normalized podocyte apoptosis and autophagy function. Further, loss of albumin (ALB) induces lipid nephrotoxicity, which we found to be alleviated by PB2 via activation of PPARα/ß-mediated lipid homeostasis and the cholesterol efflux axis. Interestingly, our results also suggested that PB2 reduces electrolyte abnormalities and edema. In addition, PB2 may contribute protective effects against trace element dys-homeostasis, which, through alleviating serum ALB loss, leads to a protective effect on glomerular permselectivity injury. Taken together, our results reveal that the identified mechanisms of PB2 on NS are multifactorial and involve inhibition of oxidative distress and inflammatory responses, as well as improvements in podocyte apoptosis and autophagy dysfunction, amelioration of lipid nephrotoxicity, and modulation of electrolyte abnormalities and edema. Thus, we provide a theoretical basis for the clinical application of PB2 against NS.
Assuntos
Nefropatias , Síndrome Nefrótica , Podócitos , Animais , Antioxidantes/metabolismo , Apoptose , Biflavonoides , Catequina , Doxorrubicina/toxicidade , Eletrólitos/efeitos adversos , Eletrólitos/metabolismo , Nefropatias/metabolismo , Lipídeos/farmacologia , Camundongos , NF-kappa B/genética , NF-kappa B/metabolismo , Síndrome Nefrótica/induzido quimicamente , Síndrome Nefrótica/tratamento farmacológico , Síndrome Nefrótica/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Podócitos/metabolismo , Proantocianidinas , Proteínas Proto-Oncogênicas c-akt/metabolismoRESUMO
Tri-(2,3-dibromopropyl) isocyanate (TBC), a new emerged persistent organic pollutant, is widely used in fields of flame retardant, textile, rubber and plastic with strong hepatotoxicity. Purple Sweet Potato Polysaccharide (PSPP) has antioxidant and hepatoprotective effects. This study aims to answer the scientific question whether PSPP has a protective effect on TBC induced liver injury. The effect of PSPP on the apoptosis of HepG2 cells was detected by MTT assay, the morphological changes were observed by morphological observation, and the apoptosis rate was determined by flow cytometry. The apoptotic genes were detected by qPCR assay, the relevant protein express was detected by western blot. The correlation between proteins and genes in the apoptosis pathway of HepG2 cells was calculated. To further reveal the apoptosis mechanism of TBC hepatotoxicity in vivo, 19 target genes and 14 apoptotic related proteins of inhibiting apoptosis via death receptor and mitochondria were discussed, all the above results proved that PSPP had protective effect on liver injury induced by TBC. This study not only provided a scientific basis for clarifying the mechanism of TBC hepatotoxicity and the protective effect of PSPP, but also generated the new point and method in terms of the prevention in advance and early intervention of diseases caused by environmental pollution.
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Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Carboidratos da Dieta/farmacologia , Ipomoea batatas/metabolismo , Isocianatos/toxicidade , Polissacarídeos/farmacologia , Células Hep G2 , HumanosRESUMO
Osteosarcoma is a commonly occurring bone malignancy, and it is the second most common cause of cancer deaths in adolescents and children. A sensitive silica nanoparticle (Si-NP) modified current-volt sensor was introduced to identify the osteopontin antigen, a well-known biomarker for osteosarcoma. Si-NP was extracted from the rice husk ash and utilized for the surface functionalization on the interdigitated microelectrode sensing surface. Extracted Si-NP has a spherical shape with uniform distribution, and it is confirmed by field emission scanning electron microscopy and field-emission transmission electron microscopy. Si-NP was layered on the electrode surface through a (3-aminopropyl)triethoxysilane amine linker, and the antibody was immobilized on Si-NP through a glutaraldehyde linker. Osteopontin was effectively detected on the antibody-attached surface, and the determination limit was 0.6 ng/mL. The regression was determined as y = 0.9366x - 1.1113 and the R2 value was 0.9331 and the detection limit of osteopontin was 0.6 ng/mL in the range between 0.3 and 5 ng/mL. In addition, control performance with nonimmune antibodies and albumin did not change the current volt, showing the specific osteopontin identification. This research work brings out the easy and cost-effective method to diagnose osteosarcoma and its etiology.
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Nanopartículas , Osteossarcoma , Adolescente , Anticorpos , Criança , Eletrodos , Humanos , Limite de Detecção , Osteopontina , Osteossarcoma/diagnóstico , Dióxido de SilícioRESUMO
OBJECTIVE: To explore the clinical early-middle stage clinical effect of percutaneous spinal endoscopic with the technique of Broad Easy Immediate Surgery(BEIS) for lumbar spinal stenosis. METHODS: The clinical data of 57 patients with lumbar spinal stenosis treated by surgery from June 2015 to June 2016 were retrospectively analyzed, including 32 cases treated by percutaneous spinal endoscopic with the technique of BEIS (minimally invasive group) and 25 cases by posterior approach of lumbar pedicle screws internal fixation and intervertebral disc excision with bone graft fusion surgery (open surgery group). The pre-operative general data such as age, Body Mass Index (BMI) and etc. were analysed and compared. The operation time, intraoperative blood loss, hospitalization time of the two groups were analyzed. Visual analogue score(VAS), Oswestry Disability Index(ODI), Japanese Orthopedic Association Score(JOA) of preoperative and at 1 week, 1 month, 3 months, 6 months after surgery were used to evaluate the clinical outcome of the two group. RESULTS: All the operations were successful and all the patients recovered smoothly without severe complications, all the wounds got good healing. The pain of the lumbar and lower limb had been improved after surgery. All the patients were followed up more than 6 months with an average of(10.65±3.38) months. There was no significant difference in general data such as age, BMI, and etc. between two groups(P>0.05). There were significant differences in VAS, ODI between two groups at 1 week, 1, 3, 6 months after surgery(P<0.05). The postoperative lumbar pain of the minimally invasive group had been better improved than the open surgery group. However, there was no significant difference in JOA between two groups at 1 week, 1, 3, 6 months after surgery(P>0.05). Intraoperative blood loss and hospitalization time of minimally invasive group was less than that of open surgery group. CONCLUSIONS: BEIS technique has the advantage of less trauma, less bleeding for lumbar stenosis when compared to open surgery. It can better alleviate the postoperative local lumbar pain, and early-middle clinical effect is equivalent to open surgery, so it can be used as a safe and effective surgical treatment for lumbar spinal stenosis.
Assuntos
Fusão Vertebral , Estenose Espinal , Humanos , Vértebras Lombares , Procedimentos Cirúrgicos Minimamente Invasivos , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The objective of this study was to identify potentially significant genes and long non-coding RNAs (lncRNAs) in colon cancer for a panel of lncRNA signatures that could be used as prognostic markers for colon adenocarcinoma (COAD) based on the data from The Cancer Genome Atlas (TCGA). RNA-seq V2 exon data of COAD were downloaded from the TCGA data portal for 285 tumor samples and 41 normal tissue samples adjacent to tumors. Differentially expressed mRNAs and lncRNAs were identified. A functional enrichment analysis of differentially expressed mRNAs was performed, followed by protein-protein interaction (PPI) network construction and significant module selection. Additionally, the regulatory relationships in differentially expressed mRNAs and lncRNAs were assessed, and an lncRNA-lncRNA co-regulation and functional synergistic analysis were performed. Furthermore, the risk score model and Cox regression analysis based on the expression levels of lncRNAs were used to develop a prognostic lncRNA signature. A total of 976 differentially expressed mRNAs and 169 differentially expressed lncRNAs were identified. MDFI and MEOX2 were the PPI network hubs. We found these lncRNAs to be mainly involved in vascular smooth muscle contraction and the cGMP-PKG signaling pathway. Several lncRNA-lncRNA pairs had co-regulatory relationships or functional synergistic effects, including BVES-AS1/MYLK-AS1, ADAMTS9-AS1/MYLK-AS1 and FENDRR/MYLK-AS1. The differential expression profile analysis of four candidate lncRNAs (MYLK-AS1, BVES-AS1, ADAMTS9-AS1, and FENDRR) in COAD tumors were confirmed by reverse transcription-quantitative PCR. Moreover, this study identified a 14-lncRNA signature that could predict the survival for COAD patients.
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Neoplasias do Colo/genética , Redes Reguladoras de Genes , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Idoso , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Transdução de Sinais , Análise de SobrevidaRESUMO
Complement dysregulation plays a key role in the pathogenesis of age-related macular degeneration (AMD), but the specific mechanisms are incompletely understood. Complement also potentiates retinal degeneration in the murine light damage model. To test the retinal function of CD59a, a complement inhibitor, CD59a knockout (KO) mice were used for light damage (LD) experiments. Retinal degeneration and function were compared in WT versus KO mice following light damage. Gene expression changes, endoplasmic reticulum (ER) stress, and glial cell activation were also compared. At baseline, the ERG responses and rhodopsin levels were lower in CD59aKO compared to wild-type (WT) mice. Following LD, the ERG responses were better preserved in CD59aKO compared to WT mice. Correspondingly, the number of photoreceptors was higher in CD59aKO retinas than WT controls after LD. Under normal light conditions, CD59aKO mice had higher levels than WT for GFAP immunostaining in Müller cells, mRNA and protein levels of two ER-stress markers, and neurotrophic factors. The reduction in photon capture, together with the neurotrophic factor upregulation, may explain the structural and functional protection against LD in the CD59aKO.
Assuntos
Antígenos CD59/genética , Luz , Células Fotorreceptoras de Vertebrados/efeitos da radiação , Degeneração Retiniana/patologia , Animais , Antígenos CD59/metabolismo , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Eletrorretinografia , Chaperona BiP do Retículo Endoplasmático , Estresse do Retículo Endoplasmático/efeitos da radiação , Células Ependimogliais/metabolismo , Enucleação Ocular , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia de Fluorescência , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Neuroglia/citologia , Neuroglia/metabolismo , Neuroglia/efeitos da radiação , Fagocitose/efeitos da radiação , Células Fotorreceptoras de Vertebrados/metabolismo , RNA Mensageiro/metabolismo , Retina/diagnóstico por imagem , Retina/metabolismo , Degeneração Retiniana/metabolismo , Degeneração Retiniana/veterinária , Retinaldeído/análise , Rodopsina/genética , Rodopsina/metabolismo , Regulação para Cima/efeitos da radiaçãoRESUMO
BACKGROUND: Colon cancer is invariably accompanied by altered coagulation activity; however, the precise role of phosphatidylserine (PS) in the hypercoagulable state of colon cancer patients remains unclear. We explored the exposure of PS on platelets and microparticles (MPs), and evaluate its role in procoagulant activity in colon cancer patients. METHODS: PS-positive platelets and MPs, mainly from platelets and endothelial cells, were detected by flow cytometry and confocal microscopy, and their procoagulant activity was assessed with purified coagulation complex assays, clotting time, and fibrin turbidity. RESULTS: Plasma levels of PS-positive platelets increased gradually from stage I to IV and were higher in all stages of the patients than in the healthy control, while PS-positive platelet-derived MPs only increased significantly in stage III/IV patients. Meanwhile, PS-positive MPs and endothelial-derived MPs in stage II/III/IV patients were markedly higher than ones in controls but no difference with stage I. Tissue factor positive MPs were higher in all 4 stages of colon cancer patients than in the healthy control. Platelets and MPs from the patients demonstrated significantly enhanced intrinsic/extrinsic FXa and thrombin generation, greatly shortened coagulation time, and increased fibrin formation. Combined treatment with PS antagonist lactadherin, strongly prolonged the coagulation time and reduced fibrin formation by inhibiting factor tenase and prothrombinase complex activity. In contrast, pretreatment with anti tissue factor antibody played a lesser role in suppression of procoagulant activity. CONCLUSION: Our results suggest that PS-positive platelets and MPs contribute to hypercoagulability and represent a potential therapeutic target to prevent coagulation in patients with colon cancer.
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Plaquetas/metabolismo , Micropartículas Derivadas de Células/metabolismo , Neoplasias do Colo/patologia , Fosfatidilserinas/farmacologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Plaquetas/patologia , Micropartículas Derivadas de Células/patologia , Neoplasias do Colo/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Trombina/metabolismo , Tromboplastina/metabolismoRESUMO
Hypercoagulability in gastric cancer is a common complication and a major contributor to poor prognosis. This study aimed to determine procoagulant activity of blood cells and microparticles (MPs) in gastric cancer patients. Phosphatidylserine-positive blood cells and MPs, and their procoagulant properties in particular, were assessed in 48 gastric cancer patients and 35 healthy controls. Phosphatidylserine-positive platelets, leukocytes, and MPs in patients with tumor-node-metastasis stage III/IV gastric cancer were significantly higher than those in stage I/II patients or healthy controls. Moreover, procoagulant activity of platelets, leukocytes, and MPs in stage III/IV patients was significantly increased compared to the controls, as indicated by shorter clotting time, higher intrinsic and extrinsic factor tenase, and prothrombinase complex activity. Interestingly, lactadherin, which competes with factors V and VIII to bind phosphatidylserine, dramatically prolonged clotting time of the cells and MPs by inhibiting factor tenase and prothrombinase complex activity. Although anti-tissue factor antibody significantly attenuated extrinsic tenase complex activity of leukocytes and MPs, it only slightly prolonged clotting times. Meanwhile, treatment with radical resection reduced phosphatidylserine-positive platelets, leukocytes, and MPs, and prolonged the clotting times of the remaining cells and MPs. Our results suggest that phosphatidylserine-positive platelets, leukocytes, and MPs contribute to hypercoagulability and represent a potential therapeutic target to prevent coagulation in patients with stage III/IV gastric cancer.
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Coagulação Sanguínea/fisiologia , Plaquetas/metabolismo , Micropartículas Derivadas de Células/metabolismo , Leucócitos/metabolismo , Fosfatidilserinas/metabolismo , Neoplasias Gástricas/sangue , Trombofilia/etiologia , Adulto , Plaquetas/fisiologia , Micropartículas Derivadas de Células/fisiologia , Feminino , Citometria de Fluxo , Humanos , Leucócitos/fisiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Neoplasias Gástricas/complicaçõesRESUMO
PURPOSE: High RPE iron levels have been associated with age-related macular degeneration. Mutation of the ferroxidase ceruloplasmin leads to RPE iron accumulation and degeneration in patients with aceruloplasminemia; mice lacking ceruloplasmin and its homolog hephaestin have a similar RPE degeneration. To determine whether a high iron diet (HID) could cause RPE iron accumulation, possibly contributing to RPE oxidative stress in AMD, we tested the effect of dietary iron on mouse RPE iron. METHODS: Male CD1 strain mice were fed either a standard iron diet (SID) or the same diet with extra iron added (HID) for either 3 months or 10 months. Mice were analyzed with immunofluorescence and Perls' histochemical iron stain to assess iron levels. Levels of ferritin, transferrin receptor, and oxidative stress gene mRNAs were measured by quantitative PCR (qPCR) in neural retina (NR) and isolated RPE. Morphology was assessed in plastic sections. RESULTS: Ferritin immunoreactivity demonstrated a modest increase in the RPE in 10-month HID mice. Analysis by qPCR showed changes in mRNA levels of iron-responsive genes, indicating moderately increased iron in the RPE of 10-month HID mice. However, even by age 18 months, there was no Perls' signal in the retina or RPE and no retinal degeneration. CONCLUSIONS: These findings indicate that iron absorbed from the diet can modestly increase the level of iron deposition in the wild-type mouse RPE without causing RPE or retinal degeneration. This suggests regulation of retinal iron uptake at the blood-retinal barriers.
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Sobrecarga de Ferro/genética , Ferro/metabolismo , Degeneração Macular/genética , Estresse Oxidativo , RNA/genética , Receptores da Transferrina/genética , Epitélio Pigmentado da Retina/metabolismo , Animais , Modelos Animais de Doenças , Sobrecarga de Ferro/induzido quimicamente , Sobrecarga de Ferro/metabolismo , Ferro da Dieta/toxicidade , Degeneração Macular/metabolismo , Degeneração Macular/patologia , Masculino , Camundongos , Camundongos Transgênicos , Reação em Cadeia da Polimerase em Tempo Real , Receptores da Transferrina/biossíntese , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/patologiaRESUMO
Brain iron accumulates in several neurodegenerative diseases and can cause oxidative damage, but mechanisms of brain iron homeostasis are incompletely understood. Patients with mutations in the cellular iron-exporting ferroxidase ceruloplasmin (Cp) have brain iron accumulation causing neurodegeneration. Here, we assessed the brains of mice with combined mutation of Cp and its homolog hephaestin. Compared to single mutants, brain iron accumulation was accelerated in double mutants in the cerebellum, substantia nigra, and hippocampus. Iron accumulated within glia, while neurons were iron deficient. There was loss of both neurons and glia. Mice developed ataxia and tremor, and most died by 9 months. Treatment with the oral iron chelator deferiprone diminished brain iron levels, protected against neuron loss, and extended lifespan. Ferroxidases play important, partially overlapping roles in brain iron homeostasis by facilitating iron export from glia, making iron available to neurons. Above: Iron (Fe) normally moves from capillaries to glia to neurons. It is exported from the glia by ferroportin (Fpn) with ferroxidases ceruloplasmin (Cp) and/or Hephaestin (Heph). Below: In mice with mutation of Cp and Heph, iron accumulates in glia, while neurons have low iron levels. Both neurons and glia degenerate and mice become ataxic unless given an iron chelator.
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Ceruloplasmina/genética , Quelantes de Ferro/uso terapêutico , Ferro/metabolismo , Proteínas de Membrana/genética , Mutação/genética , Doenças Neurodegenerativas , Piridonas/uso terapêutico , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Ceruloplasmina/metabolismo , Deferiprona , Modelos Animais de Doenças , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Transgênicos , Atividade Motora/efeitos dos fármacos , Atividade Motora/genética , Força Muscular/efeitos dos fármacos , Força Muscular/genética , Proteína Básica da Mielina/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Doenças Neurodegenerativas/tratamento farmacológico , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/metabolismo , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Gastric cancer is a lethal disease characterized by high diffusivity and mortality. To examine the mechanisms involved in gastric cancer, we analyzed the microarray of GSE41476. GSE41476 was downloaded from the Gene Expression Omnibus and included 3 primary cell culture samples from gastric cancer tissues, 3 gastric cancer cell lines and 2 normal tissue samples. Long non-coding RNAs (lncRNAs) and differentially expressed genes (DEGs) were screened by Cuffdiff software. Functions of the DEGs were predicted by functional and pathway enrichment analyses. The interaction relationships of the proteins encoded by DEGs that were obtained from the STRING database and proteinprotein interaction (PPI) network were visualized using Cytoscape. Modules analysis of PPI network was performed using CFinder. Moreover, lncRNA analysis was performed. A total of 86 lncRNAs, and 1,088 up- and 1,537 downregulated transcriptions were screened. For DEGs in module A of the PPI network for upregulated genes, the enriched pathways included ECM-receptor interaction and focal adhesion, both of which involved COL and ITG genes. The COL genes interacted with the ITG genes (e.g., COL1A1ITGA5 and COL1A2ITGB1). For DEGs in module B of the PPI network for downregulated genes, the enriched pathways for DEGs included the Tcell receptor signaling pathway, which involved PIK3CG and PIK3R5. PIK3CG had an interaction relationship with PIK3R5. In addition, IL7 was co-expressed with TCONS-00068220. In summary, the results showed that COL and ITG genes, PIK3CG, PIK3R5, IL7 and lncRNA TCONS00068220 may play a role in gastric cancer.
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Redes Reguladoras de Genes , Proteínas de Neoplasias/biossíntese , RNA Longo não Codificante/biossíntese , Neoplasias Gástricas/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Proteínas de Neoplasias/genética , RNA Longo não Codificante/genética , Transdução de Sinais , Neoplasias Gástricas/patologiaRESUMO
Dysregulation of iron homeostasis may be a pathogenic factor in age-related macular degeneration (AMD). Meanwhile, the formation of complement-containing deposits under the retinal pigment epithelial (RPE) cell layer is a pathognomonic feature of AMD. In this study, we investigated the molecular mechanisms by which complement component 3 (C3), a central protein in the complement cascade, is up-regulated by iron in RPE cells. Modulation of TGF-ß signaling, involving ERK1/2, SMAD3, and CCAAT/enhancer-binding protein-δ, is responsible for iron-induced C3 expression. The differential effects of spatially distinct SMAD3 phosphorylation sites at the linker region and at the C terminus determined the up-regulation of C3. Pharmacologic inhibition of either ERK1/2 or SMAD3 phosphorylation decreased iron-induced C3 expression levels. Knockdown of SMAD3 blocked the iron-induced up-regulation and nuclear accumulation of CCAAT/enhancer-binding protein-δ, a transcription factor that has been shown previously to bind the basic leucine zipper 1 domain in the C3 promoter. We show herein that mutation of this domain reduced iron-induced C3 promoter activity. In vivo studies support our in vitro finding of iron-induced C3 up-regulation. Mice with a mosaic pattern of RPE-specific iron overload demonstrated co-localization of iron-induced ferritin and C3d deposits. Humans with aceruloplasminemia causing RPE iron overload had increased RPE C3d deposition. The molecular events in the iron-C3 pathway represent therapeutic targets for AMD or other diseases exacerbated by iron-induced local complement dysregulation.
Assuntos
Complemento C3/metabolismo , Degeneração Macular/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Proteína delta de Ligação ao Facilitador CCAAT/metabolismo , Linhagem Celular , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Ferro/química , Sobrecarga de Ferro , Ligantes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Pessoa de Meia-Idade , Transdução de Sinais , Proteína Smad3/metabolismo , Regulação para CimaRESUMO
The retina can be shielded by the blood-retinal barrier. Because photoreceptors are damaged by excess iron, it is important to understand whether the blood-retinal barrier protects against high serum iron levels. Bone morphogenic protein 6 (Bmp6) knockout mice have serum iron overload. Herein, we tested whether the previously documented retinal iron accumulation in Bmp6 knockout mice might result from the high serum iron levels or, alternatively, low levels of retinal hepcidin, an iron regulatory hormone whose transcription can be up-regulated by Bmp6. Furthermore, to determine whether increases in serum iron can elevate retinal iron levels, we i.v. injected iron into wild-type mice. Retinas were analyzed by real-time quantitative PCR and immunofluorescence to assess the levels of iron-regulated genes/proteins and oxidative stress. Retinal hepcidin mRNA levels in Bmp6 knockout retinas were the same as, or greater than, those in age-matched wild-type retinas, indicating that Bmp6 knockout does not cause retinal hepcidin deficiency. Changes in mRNA levels of L ferritin and transferrin receptor indicated increased retinal iron levels in i.v. iron-injected wild-type mice. Oxidative stress markers were elevated in photoreceptors of mice receiving i.v. iron. These findings suggest that elevated serum iron levels can overwhelm local retinal iron regulatory mechanisms.
Assuntos
Barreira Hematorretiniana/metabolismo , Sobrecarga de Ferro/metabolismo , Ferro/metabolismo , Retina/metabolismo , Animais , Proteína Morfogenética Óssea 6/genética , Proteína Morfogenética Óssea 6/metabolismo , Hepcidinas/genética , Hepcidinas/metabolismo , Ferro/sangue , Sobrecarga de Ferro/genética , Camundongos , Camundongos Knockout , Estresse Oxidativo/fisiologiaRESUMO
PURPOSE: Oxidative stress and inflammation have key roles in the light damage (LD) model of retinal degeneration as well as in age-related macular degeneration (AMD). We sought to determine if lipoic acid (LA), an antioxidant and iron chelator, protects the retina against LD. METHODS: Balb/c mice were treated with LA or control saline via intraperitoneal injection, and then were placed in constant cool white light-emitting diode (LED) light (10,000 lux) for 4 hours. Retinas were evaluated at several time points after LD. Photoreceptor apoptosis was assessed using the TUNEL assay. Retinal function was analyzed via electroretinography (ERG). Retinal degeneration was assessed after LD by optical coherence tomography (OCT), TUNEL analysis, and histology. The mRNAs of several oxidative stress, inflammation, and iron-related genes were quantified by quantitative PCR (qPCR). RESULTS: The LD resulted in substantial photoreceptor-specific cell death. Dosing with LA protected photoreceptors, decreasing the numbers of TUNEL-positive photoreceptors and increasing the number of surviving photoreceptors. The retinal mRNA levels of genes indicating oxidative stress, inflammation, and iron accumulation were lower following LD in mice treated with LA than in control mice. The ERG analysis demonstrated functional protection by LA. CONCLUSIONS: Systemic LA is protective against light-induced retinal degeneration. Since this agent already has proven protective in other retinal degeneration models, and is safe and protective against diabetic neuropathy in patients, it is worthy of consideration for a human clinical trial against retinal degeneration or AMD.
Assuntos
Antioxidantes/farmacologia , Quelantes de Ferro/farmacologia , Retina/efeitos dos fármacos , Degeneração Retiniana/tratamento farmacológico , Ácido Tióctico/farmacologia , Animais , Apoptose/efeitos dos fármacos , Eletrorretinografia , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Marcação In Situ das Extremidades Cortadas , Luz/efeitos adversos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Retina/patologia , Degeneração Retiniana/etiologia , Degeneração Retiniana/patologia , Retinite/tratamento farmacológico , Retinite/etiologia , Retinite/patologia , Rodopsina/genética , Tomografia de Coerência Óptica , cis-trans-Isomerases/genéticaRESUMO
PURPOSE: To investigate the retinal-protective effects of the oral iron chelator deferiprone (DFP) in mice lacking the iron regulatory hormone hepcidin (Hepc). These Hepc knockout (KO) mice have age-dependent systemic and retinal iron accumulation leading to retinal degeneration. METHODS: Hepc KO mice were given DFP in drinking water from age 6 to 18 months. They were then compared to Hepc KO mice not receiving DFP by fundus imaging, electroretinography (ERG), histology, immunofluorescence, and quantitative PCR to investigate the protective effect of DFP against retinal and retinal pigment epithelial (RPE) degeneration. RESULTS: In Hepc KO mice, DFP diminished RPE depigmentation and autofluorescence on fundus imaging. Autofluorescence in the RPE layer in cryosections was significantly diminished by DFP, consistent with the fundus images. Immunolabeling with L-ferritin and transferrin receptor antibodies showed a decreased signal for L-ferritin in the inner retina and RPE cells and an increased signal for transferrin receptor in the inner retina, indicating diminished retinal iron levels with DFP treatment. Plastic sections showed that photoreceptor and RPE cells were well preserved in Hepc KO mice treated with DFP. Consistent with photoreceptor protection, the mRNA level of rhodopsin was significantly higher in retinas treated with DFP. The mRNA levels of oxidative stress-related genes heme oxygenase-1 and catalase were significantly lower in DFP-treated Hepc KO retinas. Finally, ERG rod a- and b- and cone b-wave amplitudes were significantly higher in DFP-treated mice. CONCLUSIONS: Long-term treatment with the oral iron chelator DFP diminished retinal and RPE iron levels and oxidative stress, providing significant protection against retinal degeneration caused by chronic systemic iron overload in Hepc KO mice. This indicates that iron chelation could be a long-term preventive treatment for retinal disease involving iron overload and oxidative stress.
Assuntos
Modelos Animais de Doenças , Hepcidinas/fisiologia , Quelantes de Ferro/farmacologia , Sobrecarga de Ferro/prevenção & controle , Piridonas/farmacologia , Degeneração Retiniana/prevenção & controle , Administração Oral , Animais , Deferiprona , Eletrorretinografia , Ferritinas/metabolismo , Angiofluoresceinografia , Técnica Indireta de Fluorescência para Anticorpo , Heme Oxigenase-1/genética , Quelantes de Ferro/administração & dosagem , Sobrecarga de Ferro/metabolismo , Sobrecarga de Ferro/patologia , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Imagem Óptica , Estresse Oxidativo , Piridonas/administração & dosagem , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptores da Transferrina/metabolismo , Degeneração Retiniana/metabolismo , Degeneração Retiniana/patologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/metabolismo , Epitélio Pigmentado da Retina/patologia , Rodopsina/genéticaRESUMO
OBJECTIVE: With the aim of elucidating the physiological characteristics of a low-pH tolerant strain Torulopsis glabrata RT-6. METHODS: The intracellular pH, ATP level, the membrane bound H(+)-ATPase activity, the membrane fatty acid composition and the intracellular polyphosphate content of the parent strain CCTCC M202019 and the mutant strain RT-6 were determined and compared under different pH conditions. RESULTS: Compared to that of the parent strain, the cell growth and pyruvate concentration of the mutant strain RT-6 were increased by 60.6% and 85.4% (56 h), respectively. Similarly, the strain RT-6 had higher intracellular pH compared to the control strain at external pH5.0, 4.5, and 4.0. The ATP content, the membrane bound H(+)-ATPase activity and the intracellular polyphosphate content of the mutant strain RT-6 were increased by 11.7%, 13.6%, and 3.5% at external pH 5.5, while at external pH 4.0, increased by 61%, 38.6%, and 30.8%, respectively. Furthermore, the mutant strain RT-6 exhibited higher content of the unsaturated fatty acids and higher membrane fluidity. CONCLUSION: Discharging more intracellular H+ and inhibiting the intracellular H+ production contributed to the strain RT-6's higher intracellular pH, and therefore the acid tolerance.
Assuntos
Candida glabrata/fisiologia , Trifosfato de Adenosina/metabolismo , Candida glabrata/crescimento & desenvolvimento , Candida glabrata/metabolismo , Ácidos Graxos/metabolismo , Concentração de Íons de Hidrogênio , Fluidez de Membrana/fisiologia , Polifosfatos/metabolismo , ATPases Translocadoras de Prótons/metabolismo , Ácido Pirúvico/metabolismoRESUMO
It is found that L-cysteine-capped nano-ZnS can further enhance the fluorescence intensity of the morin-nucleic acid system. Under optimum conditions, the enhanced intensity of fluorescence is proportional to the concentration of nucleic acid in the range of 7.0 x 10(-8)-1.0 x 10(-5) g mL(-1) for fish sperm DNA (fsDNA) and 9.0 x 10(-8)-5.0 x 10(-6) g mL(-1) for yeast RNA (yRNA). The corresponding detection limits (S/N = 3) are 2.0 x 10(-8) g mL(-1) and 4.0 x 10(-8) g mL(-1), respectively. The interaction mechanisms of morin-nucleic acid-L-cysteine-capped nano-ZnS system are studied by multiple techniques. It is considered that there exists synergistic effects of groove binding and electrostatic interaction between morin, L-cysteine-capped nano-ZnS and nucleic acid, and the complex of morin-L-cysteine-capped nano-ZnS-nucleic acid is formed.