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BACKGROUND: Gestational diabetes mellitus (GDM) is the most frequently occurring complication during pregnancy, with a high prevalence rate. Ferroptosis, a type of iron-dependent cell death, is closely associated with GDM nosogenesis. The present study aimed to examine the potential role and mechanism of circHIPK3 in GDM. METHODS: Placental tissues, plasma samples, and HTR-8/SVneo cells were used. A receiver operating characteristic curve was used to analyze the diagnostic value of circHIPK3 in GDM. Actinomycin D and RnaseR were added to identify circHIPK3 characteristics. The expression of circHIPK3, miR-1278, and DNA methyltransferase 1 (DNMT1) was assessed using a quantitative reverse transcriptase-PCR. Cell counting kit-8 and terminal deoxynucleotidyl transferase dUTP nick end labeling assays and specific kits were employed to assess cell viability, apoptosis, reactive oxygen species (ROS), malondialdehyde, iron, glutathione, and glutathione peroxidase 4 (GPX4) levels. RESULTS: The interaction between miR-1278 and circHIPK3 or DNMT1 was validated via luciferase reporter and RNA pull-down assays. circHIPK3 expression was found to be high in GDM placental tissues, plasma, and cells, with a high diagnostic value. In high glucose (HG)-induced HTR-8/SVneo cells, the inhibition of circHIPK3 provoked cell viability and mitigated cell apoptosis, ROS, and iron levels, but it was rescued through the downregulation of miR-1278. Mechanism experiments showed that circHIPK3 bound with miR-1278 targeting DNMT1 in GDM. The elevation in DNMT1 expression abolished the effects of miR-1278 overexpression on ferroptosis in HG-cultured HTR-8/SVneo cells. CONCLUSIONS: Overall, circHIPK3 might facilitate ferroptosis via miR-1278/DNMT1 to regulate GPX4 DNA methylation in HG-cultured HTR-8/SVneo cells. CircHIPK3 could be a therapeutic agent for GDM treatment.
Assuntos
Diabetes Gestacional , Ferroptose , MicroRNAs , Feminino , Humanos , Gravidez , Proliferação de Células/genética , Diabetes Gestacional/genética , Metilação de DNA , Ferroptose/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/genética , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/metabolismo , Placenta/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , RNA Circular/genética , RNA Circular/metabolismoRESUMO
BACKGROUND AND AIMS: Wilson's disease (WD) is a rare hereditary disorder due to ATP7B gene mutation, causing pathologic copper storage mainly in the liver and neurological systems. Hepatocyte transplantation showed therapeutic potential; however, this strategy is often hindered by a shortage of quality donor cells and by allogeneic immune rejection. In this study, we aimed to evaluate the function and efficacy of autologous reprogrammed, ATP7B gene-restored hepatocytes using a mouse model of WD. APPROACH AND RESULTS: Sufficient liver progenitor cells (LPCs) were harvested by reprogramming hepatocytes from ATP7B-/- mice with small molecules, which exhibited strong proliferation and hepatic differentiation capacity in vitro. After lentivirus-mediated mini ATP7B gene transfection and redifferentiation, functional LPC-ATP7B-derived hepatocytes (LPC-ATP7B-Heps) were developed. RNA sequencing data showed that, compared with LPC-green fluorescent protein-Heps (LPC-GFP-Heps) with enrichment of genes that were mainly in pathways of oxidative stress and cell apoptosis, in LPC-ATP7B-Heps under high copper stress, copper ion binding and cell proliferation pathways were enriched. LPC-ATP7B-Heps transplantation into ATP7B-/- mice alleviated deposition of excess liver copper with its associated inflammation and fibrosis, comparable with those observed using normal primary hepatocytes at 4 months after transplantation. CONCLUSIONS: We established a system of autologous reprogrammed WD hepatocytes and achieved ATP7B gene therapy in vitro. LPC-ATP7B-Heps transplantation demonstrated therapeutic efficacy on copper homeostasis in a mouse model of WD.
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Transplante de Células-Tronco Hematopoéticas , Degeneração Hepatolenticular , Animais , Cobre/metabolismo , ATPases Transportadoras de Cobre/genética , ATPases Transportadoras de Cobre/metabolismo , Modelos Animais de Doenças , Terapia Genética , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Hepatócitos/metabolismo , Degeneração Hepatolenticular/genética , Degeneração Hepatolenticular/metabolismo , Degeneração Hepatolenticular/terapia , MutaçãoRESUMO
Parkinson's disease (PD) is a common neurodegenerative disease. Here, the purpose of the study was to explore the function of long non-coding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) in PD and its underlying mechanism. An in vivo 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-hydrochloride (MPTP)-induced mouse model of PD was generated and the SH-SY5Y cells were treated with MPP + to induce neuronal damage in vitro. Quantitative real-time polymerase chain reaction (QRT-PCR) and Western blot were used to detect the expression of HOTAIR, miR-221-3p, α-synuclein and apoptosis-related genes. MTT, flow cytometry and TUNEL assay was used to detect cell viability and apoptosis, respectively. The levels of inflammatory cytokines TNF-α,IL-1ß and IL-6 were detected by ELISA assay. The levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS), and superoxide dismutase (SOD) were determined using the appropriate assay kits. The interactions between miR-221-3p and HOTAIR or α-synuclein were determined by dual luciferase assay and RNA binding protein immunoprecipitation (RIP). Co-localization of HOTAIR and miR-221-3p was also proved by immunofluorescence staining. The results showed that HOTAIR was highly expressed, while miR-221-3p expression was decreased in PD model in vivo and in vitro. In SH-SY5Y cells treated with MPP+, the knockdown of HOTAIR increased cell viability and reduced cell apoptosis, the secretion of inflammatory cytokines and oxidative stress reaction, while HOTAIR overexpression led to opposite effects. Furthermore, HOTAIR sponged miR-221-3p which directly targeted α-synuclein and thus regulated the expression of α-synuclein. Meanwhile, inhibiting miR-221-3p could partially reverse the neuroprotective effects of HOTAIR knockdown. In conclusion, HOTAIR attenuated the injury of SH-SY5Y cells induced by MPP+ via miR-221-3p/α-synuclein axis, suggesting the potential therapeutic value of HOTAIR in PD.
Assuntos
MicroRNAs , Doenças Neurodegenerativas , Doença de Parkinson , RNA Longo não Codificante/metabolismo , alfa-Sinucleína/metabolismo , Animais , Apoptose , Linhagem Celular Tumoral , Humanos , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Doença de Parkinson/metabolismo , Doença de Parkinson/patologiaRESUMO
BACKGROUND: Patients with gynecologic cancer receiving chemotherapy often report unmet supportive care needs. Compared with traditional face-to-face clinical interventions, mobile health can increase access to supportive care and may address patients' needs. Although app-based support programs have been developed to support patients with gynecologic cancer, their efficacy has not been adequately tested. OBJECTIVE: The aim of this study was to examine the efficacy of a mobile app for gynecologic cancer support (MGCS) for patients with gynecologic cancer receiving chemotherapy in China. METHODS: A multicenter randomized controlled trial was conducted in 2 university-affiliated hospitals in China. A total of 168 Chinese patients with gynecologic cancer were recruited and randomized to receive routine care or MGCS program plus routine care for 24 weeks. The Mishel uncertainty in illness theory guided the development of MGCS program, which has 4 modules: weekly topics, emotional care, discussion center, and health consultation. The primary outcome of this program was the assessment of the uncertainty in illness. The secondary outcomes were quality of life, symptom distress, and social support. All health outcomes were evaluated at baseline (T0), 12 weeks (T1), and 24 weeks (T2). Repeated measures analysis of covariance was used to assess the efficacy of the MGCS program. RESULTS: In this trial, 67 patients in the control group and 69 patients in the intervention group completed 2 follow-up assessments (response rate, 136/168, 81%). At 12 weeks, no significant differences were observed in any of the health outcomes between the 2 groups. At 24 weeks, compared to patients in the control group, those in the intervention group reported significant decreased uncertainty in illness (P<.001; d=-0.60; adjusted mean difference -7.69, 95% CI -11.31 to -4.07) and improved quality of life (P=.04; d=0.30; adjusted mean difference 4.77, 95% CI 0.12-9.41). CONCLUSIONS: The MGCS program demonstrated efficacy in supporting patients with gynecologic cancer receiving chemotherapy. This trial illustrates that an app-based program can be incorporated into routine care to support patients with cancer and suggests that allocation of more resources (grants, manpower, etc) to mobile health in clinics is warranted. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2000033678; https://www.chictr.org.cn/showproj.html?proj=54807.
Assuntos
Neoplasias dos Genitais Femininos , Aplicativos Móveis , Feminino , Humanos , China , Neoplasias dos Genitais Femininos/tratamento farmacológico , Qualidade de Vida , Apoio SocialRESUMO
AIMS: This study was to assess the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) in inpatient settings. DESIGN: A case-control study. METHODS: This study was conducted in a tertiary hospital based on 2019 data. With a case-control design in a 1:2 ratio, the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) was determined by ROC curve. The best cut point was identified based on sensitivity, specificity, positive predict value and negative predict value. Conditional logistical regression analysis was conducted to test the predictive ability of each indicator. RESULTS: The study included 309 patients, with 103 in the case group and 206 in the control groups. Generally, the predictive ability was acceptable with the area under ROC curve value at 0.73 (95% CI: 0.67-0.79). Positive predict value and negative predict value performed best at the cut point of 13. Sensitivity at cut point 6 was much higher than that at cut point 13, though specificity was lower. Except for age, all indicators in the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) demonstrated significant predictive ability as to occurrence of fall. CONCLUSION: The Johns Hopkins Fall Risk Assessment Tool (Chinese Version) is a reliable assessment instrument in the inpatient settings. IMPACT: This is the first study that evaluated the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese version) in the inpatient settings, and proved that the instrument is reliable for assessing inpatient fall risks. Further studies could be carried out to assess the predict ability of Johns Hopkins Fall Risk Assessment Tool (Chinese version) among specific populations.
Assuntos
Pacientes Internados , Humanos , Estudos de Casos e Controles , Medição de Risco , ChinaRESUMO
Shear stress was reported to regulate the expression of AC007362, but its underlying mechanisms remain to be explored. In this study, to isolate endothelial cells of blood vessels, unruptured and ruptured intracranial aneurysm (IA) tissues were collected from IA patients. Subsequently, quantitative real-time PCR (qRT-PCR), Western blot and luciferase assay were performed to investigate the relationships between AC007362, miRNAs-493 and monocyte chemoattractant protein-1 (MCP-1) in human umbilical vein endothelial cells (HUVECs) exposed to shear stress. Reduced representation bisulphite sequencing (RRBS) was performed to assess the level of DNA methylation in AC007362 promoter. Accordingly, AC007362 and MCP-1 were significantly up-regulated while miR-493 was significantly down-regulated in HUVECs exposed to shear stress. AC007362 could suppress the miR-493 expression and elevate the MCP-1 expression, and miR-493 was shown to respectively target AC007362 and MCP-1. Moreover, shear stress in HUVECs led to the down-regulated DNA methyltransferase 1 (DNMT1), as well as the decreased DNA methylation level of AC007362 promoter. Similar results were also observed in ruptured IA tissues when compared with unruptured IA tissues. In conclusion, this study presented a deep insight into the operation of the regulatory network of AC007362, miR-493 and MCP-1 upon shear stress. Under shear stress, the expression of AC007362 was enhanced by the inhibited promoter DNA methylation, while the expression of MCP-1 was enhanced by sponging the expression of miR-493.
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Quimiocina CCL2/metabolismo , Células Endoteliais da Veia Umbilical Humana/patologia , Aneurisma Intracraniano/patologia , Reologia , Estresse Mecânico , Sequência de Bases , Quimiocina CCL2/genética , Metilação de DNA/genética , Feminino , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Aneurisma Intracraniano/genética , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/genética , Regulação para Cima/genéticaRESUMO
The genus Amolops ("torrent frogs") is one of the most species-rich genera in Ranidae, with 59 recognized species. This genus currently includes six species groups diagnosed mainly by morphology. Several recent molecular studies indicated that the classification of species groups within Amolops remains controversial, and key nodes in the phylogeny have been inadequately resolved. In addition, the diversity of Amolops remains poorly understood, especially for those from incompletely sampled regions. Herein, we investigate species-level diversity within the genus Amolops throughout southern China and Southeast Asia, and infer evolutionary relationships among the species using mtDNA data (16S, COI, and ND2). Molecular analyses indicate nine unnamed species, mostly distributed in the Himalayas. We then utilized anchored hybrid enrichment to generate a dataset representing the major mitochondrial lineages to resolve phylogenetic relationships, biogeography, and pattern of species diversification. Our resulting phylogeny strongly supports the monophyly of four previously identified species groups (the A. ricketti, A. daiyunensis, A. hainanensis, and A. monticola groups), but paraphyly for the A. mantzorum and A. marmoratus groups, as previously defined. We erect one new species group, the A. viridimaculatus group, and recognize Dubois' (1992) subgenus Amo as the A. larutensis species group. Biogeographic analysis suggests that Amolops originated on the Indo-Burma/Thai-Malay Peninsula at the Eocene/Oligocene boundary, and dispersed outward, exemplifying a common pattern observed for the origin of Asian biodiversity. The early divergence within Amolops coincides with the Himalayan uplift and the lateral extrusion of Indochina at the Oligocene/Miocene boundary. Our results show that paleoclimatic and geomorphological events have profoundly influenced the patterns of lineage diversification within Amolops.
Assuntos
Biodiversidade , Núcleo Celular/genética , DNA Mitocondrial/genética , Filogenia , Filogeografia , Ranidae/genética , Animais , Sudeste Asiático , Sequência de Bases , Teorema de Bayes , Análise Espaço-Temporal , Especificidade da EspécieRESUMO
Parkinson's disease (PD) is neurodegenerative dyskinesia characterized by loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Although neuroinflammation is one of the pathological features of PD, its mechanism of promoting PD is still not fully understood. Recently, the microRNA (miR) is considered to play a critical regulatory role in inflammatory responses. In this study, we examined the anti-inflammatory activity, antineuronal injury, and the underlying target of miR-190 with MPTP-induced PD mouse model and BV2 cells. The results showed that miR-190 is downregulated in lipopolysaccharide (LPS)-induced BV2 cells; however, when the miR-190 overexpressed, the expression of proinflammatory mediators, such as iNOS, IL-6, TNF-α, and TGF-ß1, were inhibited and the anti-inflammatory mediator such IL-10 was increased. In addition, we predicted the potential target of miR-190 to be Nlrp3 and verified by luciferase reporter assay. The results also showed that Nlrp3 was upregulated in LPS-induced BV2 cells, whereas knockdown of Nlrp3 inhibited the LPS-induced inflammatory response in BV2 cells. Furthermore, upregulation of miR-190 or knockdown of Nlrp3 inhibited LPS-induced apoptosis in BV2 cells. However, the apoptosis inhibition effect of miR-190 was abrogated by overexpression of Nlrp3. Finally, upregulation of miR-190 inhibited the activation of microglial cells and inflammation and attenuated the tyrosine hydroxylase loss in SNpc in MPTP-induced PD mice. In conclusion, we demonstrated that miR-190 alleviates neuronal damage and inhibits inflammation via negatively regulating the expression and activation of Nlrp3 in MPTP-induced PD mouse model.
Assuntos
Apoptose , Encéfalo/metabolismo , Encefalite/metabolismo , Inflamassomos/metabolismo , Intoxicação por MPTP/metabolismo , MicroRNAs/metabolismo , Microglia/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Neurônios/metabolismo , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina , Animais , Encéfalo/patologia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Encefalite/induzido quimicamente , Encefalite/genética , Encefalite/patologia , Humanos , Inflamassomos/genética , Mediadores da Inflamação/metabolismo , Intoxicação por MPTP/induzido quimicamente , Intoxicação por MPTP/genética , Intoxicação por MPTP/patologia , Masculino , Camundongos Endogâmicos C57BL , MicroRNAs/genética , Microglia/patologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Neurônios/patologia , Transdução de SinaisRESUMO
Matrix metalloproteinase 9 (MMP-9) plays a beneficial role in the delayed phase of middle cerebral artery occlusion (MCAO). However, the mechanism is obscure. Here, we constructed hypoxia response element (HRE)-regulated MMP-9 to explore its effect on glial scars and neurogenesis in delayed ischemic stroke. Adult male Institute of Cancer Research (ICR) mice underwent MCAO and received a stereotactic injection of lentivirus carrying HRE-MMP-9 or normal saline (NS)/lentivirus-GFP 7 days after ischemia. We found that HRE-MMP-9 improved neurological outcomes, reduced ischemia-induced brain atrophy, and degraded glial scars (p < 0.05). Furthermore, HRE-MMP-9 increased the number of microvessels in the peri-infarct area (p < 0.001), which may have been due to the accumulation of endogenous endothelial progenitor cells (EPCs) in the peri-infarct area after glial scar degradation. Finally, HRE-MMP-9 increased the number of bromodeoxyuridine-positive (BrdU+)/NeuN+ cells and the expression of PSD-95 in the peri-infarct area (p < 0.01). These changes could be blocked by vascular endothelial growth factor receptor 2 (VEGFR2) inhibitor SU5416 and MMP-9 inhibitor 2-[[(4-phenoxyphenyl)sulfonyl]methyl]-thiirane (SB-3CT). Our results provided a novel mechanism by which glial scar degradation and vascular endothelial growth factor (VEGF)/VEGFR2-dependent angiogenesis may be key procedures for neurological recovery in delayed ischemic stroke after HRE-MMP-9 treatment. Therefore, HRE-MMP-9 overexpression in the delayed ischemic brain is a promising approach for neurological recovery.
Assuntos
Hipóxia/genética , Hipóxia/metabolismo , Metaloproteinase 9 da Matriz/genética , Neovascularização Fisiológica/genética , Neuroglia/metabolismo , Elementos de Resposta , Acidente Vascular Cerebral/genética , Acidente Vascular Cerebral/metabolismo , Animais , Astrócitos/metabolismo , Atrofia , Encéfalo/metabolismo , Encéfalo/patologia , Movimento Celular , Modelos Animais de Doenças , Matriz Extracelular/metabolismo , Infarto da Artéria Cerebral Média , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Neurogênese/genética , Neuroglia/patologia , Neurônios/metabolismo , Acidente Vascular Cerebral/patologia , Acidente Vascular Cerebral/fisiopatologia , Reabilitação do Acidente Vascular CerebralRESUMO
The induced pluripotent stem cell (iPSC) technology has transformed in vitro research and holds great promise to advance regenerative medicine. iPSCs have the capacity for an almost unlimited expansion, are amenable to genetic engineering, and can be differentiated into most somatic cell types. iPSCs have been widely applied to model human development and diseases, perform drug screening, and develop cell therapies. In this review, we outline key developments in the iPSC field and highlight the immense versatility of the iPSC technology for in vitro modeling and therapeutic applications. We begin by discussing the pivotal discoveries that revealed the potential of a somatic cell nucleus for reprogramming and led to successful generation of iPSCs. We consider the molecular mechanisms and dynamics of somatic cell reprogramming as well as the numerous methods available to induce pluripotency. Subsequently, we discuss various iPSC-based cellular models, from mono-cultures of a single cell type to complex three-dimensional organoids, and how these models can be applied to elucidate the mechanisms of human development and diseases. We use examples of neurological disorders, coronavirus disease 2019 (COVID-19), and cancer to highlight the diversity of disease-specific phenotypes that can be modeled using iPSC-derived cells. We also consider how iPSC-derived cellular models can be used in high-throughput drug screening and drug toxicity studies. Finally, we discuss the process of developing autologous and allogeneic iPSC-based cell therapies and their potential to alleviate human diseases.
Assuntos
COVID-19 , Reprogramação Celular , Células-Tronco Pluripotentes Induzidas , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Reprogramação Celular/genética , SARS-CoV-2/genética , Diferenciação Celular/genética , Neoplasias/genética , Neoplasias/terapia , Neoplasias/patologia , Medicina Regenerativa , Doenças do Sistema Nervoso/terapia , Doenças do Sistema Nervoso/genética , Doenças do Sistema Nervoso/patologiaRESUMO
Stroke is a leading cause of disability and mortality all over the world. Due to an aging population, the incidence of stroke is rising significantly, which has led to devastating consequences for patients. In addition to traditional risk factors such as age, hypertension, hyperlipidemia, diabetes and atrial fibrillation, sleep disorders, as independent modifiable risk factors for stroke, have been highlighted increasingly. In this review, we provide an overview of common types of current sleep disturbances in cerebrovascular diseases, including insomnia, hypersomnia, breathing-related sleep disorders, and parasomnias. Moreover, evidence-based clinical therapeutic strategies and pitfalls of specific sleep disorders after stroke are discussed. We also review the neurobiological mechanisms of these treatments as well as their effects on stroke. Since depression after stroke is so prevalent and closely related to sleep disorders, treatments of post-stroke depression are also briefly mentioned in this review article.
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Pelvic organ prolapse is a worldwide health problem to elderly women. Understanding its pathogenesis and an ideal animal model are crucial to developing promising treatments. The present study aimed to investigate new clinical significance and detailed mechanism of pelvic organ prolapse by comparing the structural, functional and molecular dysfunctions of pelvic organ prolapse in patient and Loxl1 deficient mice. Our results showed that human vagina tissues from prolapsed site showed disarranged collagen and elastic fibers compared with the non-prolapse tissue. A gene ontology (GO) analysis of differentially expressed genes revealed molecular changes mainly related to inflammatory response and extracellular matrix (ECM) organization. While the mice lacking Loxl1 developed stable POP phenotype and disordered ECM structure in histology. Such Loxl1 knockout mice exhibited a significantly urinary dysfunction and decreased mechanical properties of the pelvic floor tissues, implying that POP in human condition might be induced by progressively decreased mechanics of pelvic tissues following ECM catabolism. Similarly, we not only identified significant up-regulated ECM catabolism processes and down-regulated ECM synthesis processes, but also characterized high level of inflammatory response in vagina tissue of the Loxl1 deficient mice. Thus, all these pathological changes in the POP mice model was consistent with those of the clinical elderly patients. These findings provide new insight into remodeling of POP by LOXL1 regulation and be of great importance to develop combination treatments of ECM metabolism and inflammation regulation strategy.
Assuntos
Aminoácido Oxirredutases/genética , Ontologia Genética , Prolapso de Órgão Pélvico , Idoso , Idoso de 80 Anos ou mais , Animais , Colágeno/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pessoa de Meia-Idade , Prolapso de Órgão Pélvico/genética , Prolapso de Órgão Pélvico/fisiopatologiaRESUMO
Long non-coding RNA (lncRNA) plays a crucial role in multiple disorders, while the role of it in Parkinson's disease (PD) is still unclear. Here, the increased lncRNA NEAT1 was discovered in MPP+-induced SH-SY5Y cells. Then, we proved that NEAT1 decreasing suppressed MPP+-induced neuronal apoptosis, upregulation of α-syn and activation of NLRP3 inflammasome. Rescue experiments shown that the inhibition of NEAT1 decreasing to MPP+-induced activation of NLRP3 inflammasome and subsequent neuronal apoptosis can be reversed by overexpressed α-syn. Subsequently, we indicated the interaction between NEAT1 and miR-1301-3p, as well as between NEAT1 and miR-5047. Interesting, we found that NEAT1 decreasing repressed the expression of GJB1, a downstream target of miR-1301-3p and miR-5047, through promoting miR-1301-3p rather than miR-5047 expression. Finally, we transfected miR-1301-3p inhibitor to MPP+-induced SH-SY5Y cells following si-NEAT1, and found that downregulation of NEAT1 repressed α-syn-mediated the activation of NLRP3 inflammasome through regulating miR-1301-3p/GJB1 signaling pathway. Overall, our data demonstrated that NEAT1 decreasing effectively suppressed MPP+-induced neuronal apoptosis. Mechanismly, downregulation of NEAT1 repressed α-syn-induced activation of NLRP3 inflammasome via inhibiting the expression of GJB1 by targeting miR-1301-3p. Our study supported a new and reliable evidence for lncRNA NEAT1 as a potential target for PD treatment.
Assuntos
MicroRNAs/antagonistas & inibidores , Doença de Parkinson/genética , RNA Longo não Codificante/genética , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/biossíntese , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular , Conexinas/biossíntese , Conexinas/genética , Progressão da Doença , Regulação para Baixo , Regulação da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Inflamassomos/fisiologia , MicroRNAs/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Proteína 3 que Contém Domínio de Pirina da Família NLR/biossíntese , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , RNA Longo não Codificante/biossíntese , Transdução de Sinais , alfa-Sinucleína/fisiologia , Proteína beta-1 de Junções ComunicantesRESUMO
Severe infection and mechanical injury of the uterus may lead to infertility and miscarriage. Currently, there is a lack of effective treatment modality for functional repair of uterine injury. To address this clinical challenge, this study aimed to develop a chemotactic composite scaffold by incorporating recombinant human stromal cell-derived factor-1α (rhSDF-1α) into a silk fibroin-bacterial cellulose (SF-BC) membrane carrier. A rat model of uterine injury was utilized for this study, which was composed of three groups as follows: blank control, implantation with SF-BC only, or SF-BC loaded with rhSDF-1α. The tissue regeneration efficacy of the three groups was analyzed and compared. The results showed that SF-BC loaded with rhSDF-1α significantly enhanced endometrial regeneration and arteriogenesis of the injured rat uterus, which led to improved pregnancy outcomes, thus indicating much promise for functional uterine repair and regeneration. Impact Statement In this study, we demonstrated that the silk fibroin-bacterial cellulose (SF-BC) membrane possessed good physical, chemical, and biocompatibility properties in vitro. The in vivo study showed that the incorporation of recombinant human stromal cell-derived factor-1α (rhSDF-1α) within the SF-BC membrane promoted regeneration of full-thickness uterine injury and also improved the pregnancy outcome of the damaged uterus. The results thus suggest that SF-BC loaded with rhSDF-1α has good potential in future clinical applications for the repair of uterine injury.
Assuntos
Celulose/química , Quimiocina CXCL12/administração & dosagem , Quimiocina CXCL12/farmacologia , Fibroínas/química , Regeneração/efeitos dos fármacos , Útero/patologia , Animais , Artérias/efeitos dos fármacos , Artérias/crescimento & desenvolvimento , Endométrio/fisiologia , Células Epiteliais/efeitos dos fármacos , Feminino , Humanos , Masculino , Membranas Artificiais , Organogênese/efeitos dos fármacos , Gravidez , Resultado da Gravidez , Ratos Sprague-Dawley , Útero/efeitos dos fármacos , Útero/cirurgiaRESUMO
Matrix metalloproteinase-9 (MMP-9) plays a beneficial role in the sub-acute phase after ischemic stroke. However, unrestrained MMP-9 may disrupt the blood-brain barrier (BBB), which has limited its use for the treatment of brain ischemia. In the present study, we constructed lentivirus mediated hypoxia-controlled MMP-9 expression and explored its role after stroke. Hypoxia response element (HRE) was used to confine MMP-9 expression only to the hypoxic region of mouse brain after 120-min transient middle cerebral artery occlusion. Lentiviruses were injected into the peri-infarct area on day 7 after transient ischemia. We found hyperexpression of exogenous HRE-MMP-9 under the control of hypoxia, and its expression was mainly located in neurons and astrocytes without aggravation of BBB damage compared to the CMV group. Furthermore, mice in the HRE-MMP-9 group showed the best behavioral recovery compared with the normal saline, GFP, and SB-3CT groups. Therefore, hypoxia-controlled MMP-9 hyperexpression during the sub-acute phase of ischemia may provide a novel promising approach of gene therapy for stroke.
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Barreira Hematoencefálica/metabolismo , Hipóxia-Isquemia Encefálica/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Animais , Atrofia , Vetores Genéticos/metabolismo , Células HEK293 , Compostos Heterocíclicos com 1 Anel/farmacologia , Humanos , Fator 1 Induzível por Hipóxia/metabolismo , Lentivirus/fisiologia , Camundongos , Atividade Motora , Recuperação de Função Fisiológica , Elementos de Resposta , Teste de Desempenho do Rota-Rod , Sulfonas/farmacologia , Junções Íntimas/metabolismoRESUMO
Ischemic stroke is a leading cause of mortality and morbidity worldwide. Numerous therapeutic studies, including stem cell therapy, have been explored in order to find the possible approach treating cerebral ischemia effectively. However, massive grafted-cell death and low survival rate made us pessimistic about the widespread application of stem cell therapy. The approach of preconditioning stem cells became an attractive option due to its high cell viability after transplantation. In this review, we focused on preconditioning stem cells via cytokines, chemical drugs and other factors including interleukin 6 (IL-6), lipopolysaccharide (LPS), minocycline, melatonin and hypoxia to increase grafted stem cell survival, proliferation, migration to ischemic peri-focal area and ultimately improve neurobehavioral outcomes. We explored the underlying therapeutic efficacy of preconditioning stem cells as well as possible mechanisms and potential risks. Finally, we discussed the prospects of the application of cell-based therapy in clinical patients with ischemic stroke.