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INTRODUCTION: This study aimed to investigate the characteristics of retinal vascular degeneration and the expression of vessel-related claudin (CLD) proteins in retinal degeneration mouse (Pde6ßrd1/rd1 rd1 mouse). METHODS: Retinas from wild-type (WT) mice and rd1 mice at postnatal day 3 (P3), P5, P8, P11, P13, P15, P18, and P21 were collected. Immunofluorescence staining was used to assess the retinal vascular plexus, cell proliferation, CLD expression, and retinal ganglion cells (RGCs). The distribution of retinal superficial and deep vessels was determined by isolectin B4 fluorescence staining of retinal flat mounts and frozen sections. Hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated dNTP nick-end labeling were used to investigate retinal histological degeneration and apoptosis in rd1 mice, respectively. Quantitative real-time PCR and Western blot were used to measure the expression of vessel-related CLD-1, -2, -3, and -5, vascular endothelial growth factor A (VEGFA), and vascular endothelial growth factor receptor 2 (VEGFR2) in the retinas. RESULTS: Compared to the WT mice, the rd1 mice displayed delayed but completed progressive development in the retinal superficial vascular plexuses (SVPs) and deep vascular plexuses (DVPs). In the rd1 mice, the thickness of retinal layers gradually decreased and the retinas underwent progressive atrophy and degeneration. The deterioration got worse at the late developmental stage. The declined vessel density of SVP and DVP correlated with the decreased thickness of the full and inner parts of the retina and the reduced number of RGCs. DVP degeneration and the thinning of the outer nuclear layer exhibited an obvious reduction at P15. The expression levels of CLD-1, CLD-2, CLD-3, CLD-5, VEGFA, and VEGFR2 decreased and were consistently lower in the rd1 mice than in WT mice since P15. CONCLUSION: Rd1 mice exhibited progressive vascular degeneration of retinal SVP and DVP, the thinning and atrophy of retinal ONL and RGC, and the downregulation of vessel-related CLD proteins during the late developmental period. Thus, the rd1 mouse is a useful model of not only retinal neuro-degeneration but also retinal vascular degeneration.
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Western Blotting , Claudinas , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , Degeneración Retiniana , Células Ganglionares de la Retina , Vasos Retinianos , Animales , Ratones , Degeneración Retiniana/metabolismo , Degeneración Retiniana/patología , Degeneración Retiniana/genética , Vasos Retinianos/patología , Vasos Retinianos/metabolismo , Claudinas/genética , Claudinas/metabolismo , Claudinas/biosíntesis , Células Ganglionares de la Retina/patología , Células Ganglionares de la Retina/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Apoptosis , Proliferación Celular , Etiquetado Corte-Fin in Situ , Regulación de la Expresión Génica , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genéticaRESUMEN
The study aimed to investigate the role of microRNA (miR)-124-3p in retinal angiogenesis in a mouse model. An intravitreal injection of miR-124-3p antagomir was used to knockdown the expression of miR-124-3p in the mouse retina at postnatal day (P)3. Immunofluorescent staining of both retinal frozen sections and whole retina were used to observe retinal vascular development in the P6, P9 and P12 mice, as well as the changes in retinal ganglion cells, astrocytes, Müller cells and microglia. Whole retinal RNA extracted from P9 mice was used for transcriptome sequencing. Following gene set enrichment analysis, the enriched genes caused by miR-124-3p inhibition were analyzed by immunofluorescent staining and western blot. Results indicated that deep vascular development was significantly inhibited by the activation of M1 phenotype microglia. Moreover, there were no notable effects on superficial retinal vascular development, the retinal ganglion cells, astrocytes, and Müller cells. The expression of the Stat1/Irf9/Eif2ak2/Ripk1 axis in the miR-124-3p knockdown group was significantly increased. The microglia penetrated deep into the retina and the activation of Ripk1(+) microglia significantly increased, which was accompanied by an increased level of apoptosis to inhibit the deep vascular sprout. Downregulation of miR-124-3p during the early retinal development can suppress the development of the deep retinal blood vessels by enhancing the expression level of the Stat1/Irf9/Eif2ak2/Ripk1 axis and inducing the cell apoptosis of the activation of Ripk1(+) microglia.
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MicroARNs , Microglía , Ratones , Animales , Regulación hacia Abajo , Microglía/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Retina/metabolismo , Vasos Retinianos/metabolismo , Apoptosis/genética , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT1/metabolismo , Proteína Serina-Treonina Quinasas de Interacción con ReceptoresRESUMEN
BACKGROUND: The present study was designed to explore the potential regulatory mechanism between mitophagy and pyroptosis during sepsis-associated acute lung injury (ALI). METHODS: In vitro or in vivo models of sepsis-associated ALI were established by administering lipopolysaccharide (LPS) or performing caecal ligation and puncture (CLP) surgery. Pyroptosis levels were detected by electron microscopy, immunofluorescence, flow cytometry, western blotting and immunohistochemistry. Dual-luciferase reporter gene assay was applied to verify the targeting relationship between miR-138-5p and NLRP3. Methylation-specific PCR and chromatin immunoprecipitation assays were used to determine methylation of the miR-138-5p promoter. Mitophagy levels were examined by transmission electron microscopy and western blotting. RESULTS: NLRP3 inflammasome silencing alleviated alveolar macrophage (AM) pyroptosis and septic lung injury. In addition, we confirmed the direct targeting relationship between miR-138-5p and NLRP3. Overexpressed miR-138-5p alleviated AM pyroptosis and the pulmonary inflammatory response. Moreover, the decreased expression of miR-138-5p was confirmed to depend on promoter methylation, while inhibition of miR-138-5p promoter methylation attenuated AM pyroptosis and pulmonary inflammation. Here, we discovered that an increased cytoplasmic mtDNA content in sepsis-induced ALI models induced the methylation of the miR-138-5p promoter, thereby decreasing miR-138-5p expression, which may activate the NLRP3 inflammasome and trigger AM pyroptosis. Mitophagy, a form of selective autophagy that clears damaged mitochondria, reduced cytoplasmic mtDNA levels. Furthermore, enhanced mitophagy might suppress miR-138-5p promoter methylation and relieve the pulmonary inflammatory response, changes that were reversed by treatment with isolated mtDNA. CONCLUSIONS: In summary, our study indicated that mitophagy induced the demethylation of the miR-138-5p promoter, which may subsequently inhibit NLRP3 inflammasome, AM pyroptosis and inflammation in sepsis-induced lung injury. These findings may provide a promising therapeutic target for sepsis-associated ALI.
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Lesión Pulmonar Aguda , MicroARNs , Sepsis , Humanos , Inflamasomas/metabolismo , Piroptosis , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Mitofagia , Lesión Pulmonar Aguda/genética , Lesión Pulmonar Aguda/metabolismo , Sepsis/complicaciones , Sepsis/genética , Desmetilación , Lipopolisacáridos/farmacología , Lipopolisacáridos/metabolismoRESUMEN
INTRODUCTION: Vascular endothelial cell injury and angiogenesis induced by hyperglycemia are the main pathological basis of vascular complications in diabetes mellitus. Our study aimed to investigate the role and mechanism of miR-210-3p in high glucose (HG)-induced angiogenesis. METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with HG to mimic the pathological process of hyperglycemia. HUVECs were divided into the control group, HG group, HG+inhibitor-NC group, and HG+miR-210-3p inhibitor group. Proliferation and migration were tested by wound healing assay, tube formation, and Transwell assay. Quantitation real-time PCR and Western blots were performed to determine the expression of miR-210-3p and relative proteins, respectively. RESULTS: The level of miR-210-3p significantly increased in HUVECs treated by HG. The knockdown of miR-210-3p attenuated the tube formation, proliferation, and migration of cultured HUVECs in vitro to inhibit angiogenesis by increasing the expression of fibroblast growth factor receptor-like 1 (FGFRL1) and then attenuating the phosphorylation of signal transducer and activator of transcription 3 (STAT3), extracellular regulated protein kinases, and protein kinase B (Akt). CONCLUSION: Our study revealed that miR-210-3p might be a promising target for treating diabetic-associated vascular injury.
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Diabetes Mellitus , Hiperglucemia , MicroARNs , Humanos , Regulación hacia Abajo , MicroARNs/genética , Angiogénesis , Células Endoteliales de la Vena Umbilical Humana , Diabetes Mellitus/metabolismo , Hiperglucemia/genética , Hiperglucemia/metabolismo , Glucosa/toxicidad , Proliferación Celular , Receptor Tipo 5 de Factor de Crecimiento de Fibroblastos/metabolismoRESUMEN
MicroRNA (miRNA) is a non-coding RNA that can regulate the expression of many target genes, and it is widely involved in various important physiological activities. MiR-124-3p was found to associate with the normal development of retinal vessels in our previous study, but the mechanism of its anti-angiogenic effect on pathological retinal neovascularization still needed to be explored. Therefore, this study aimed to investigate the effect and mechanism of miR-124-3p on retinal neovascularization in mice with oxygen-induced retinopathy (OIR). Here, we found that intravitreal injection of miR-124-3p agomir attenuated pathological retinal neovascularization in OIR mice. Moreover, miR-124-3p preserved the astrocytic template, inhibited reactive gliosis, and reduced the inflammatory response as well as necroptosis. Furthermore, miR-124-3p inhibited the signal transducer and activator of transcription 3 (STAT3) pathway and decreased the expression of hypoxia-inducible factor-1α and vascular endothelial growth factor. Taken together, our results revealed that miR-124-3p inhibited retinal neovascularization and neuroglial dysfunction by targeting STAT3 in OIR mice.
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MicroARNs , Neovascularización Retiniana , Animales , Ratones , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , MicroARNs/genética , MicroARNs/metabolismo , Neuroglía/metabolismo , Oxígeno/efectos adversos , Oxígeno/metabolismo , Neovascularización Retiniana/metabolismo , Factor de Transcripción STAT3/genética , Factor de Transcripción STAT3/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND: The prevalence and intensity of menopausal symptoms differ depending on ethnicity, culture, and country. Epidemiological data from China are scarce. OBJECTIVE: To compare the prevalence and severity of menopausal symptoms in peri- and postmenopausal Chinese women. METHODS: This was a prospective two year cohort study that included all eligible women from 31 Chinese provinces attending our 'Menopause Clinic', the first official specialized center in China. Structured questionnaires containing seven domains with 41 items in total were used to assess the following menopausal symptoms using descriptive analysis: negative mood, cognitive symptoms, sleep disorder, vasomotor symptoms (VMS), urogenital symptoms, autonomic nervous disorder, and limb pain/paresthesia. RESULTS: A total of 4063 women with a mean age of 50.53 ± 6.57 (n = 2107 perimenopausal and 1956 postmenopausal) participated. All menopausal symptoms were more severe in postmenopausal women (p<.05). Independent of menopausal status, urogenital symptoms, often combined with sexual problems, were the most common complaints (in prevalence and severity), followed by sleep disorder, cognitive symptoms (especially hypomnesia), negative mood, autonomic nervous disorder, limb pain/paresthesia and, as the rarest complaint, VMS. CONCLUSIONS: Urogenital symptoms among midlife Chinese women are common, frequently also in combination with sexual dysfunction, although many do not often complain about these in the first place. Postmenopausal women presented more prevalent and severe menopausal symptoms. In contrast to Western countries, VMS are rare among our population. A multidisciplinary approach and use of hormonal and non-hormonal therapies should be considered for these women.
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Perimenopausia/fisiología , Posmenopausia/fisiología , Adulto , Pueblo Asiatico/estadística & datos numéricos , China/epidemiología , Femenino , Humanos , Persona de Mediana Edad , Perimenopausia/etnología , Perimenopausia/psicología , Posmenopausia/etnología , Posmenopausia/psicología , Prevalencia , Estudios ProspectivosRESUMEN
Diabetic retinopathy (DR) is the most common complication of diabetes. Proliferative DR (PDR) is a more advanced stage of DR, which can cause severe impaired vision and even blindness. However, the precise pathological mechanisms of PDR remain unknown. DNA methylation serves an important role in the initiation and progression of numerous types of disease including PDR. The purpose of this study was to identify the aberrantly methylated differentially expressed genes (DEGs) as potential therapeutic targets of PDR. The gene expression microarray dataset GSE60436 and the methylation profiling microarray dataset GSE57362 were used to determine the aberrantly methylated DEGs in PDR, utilizing normal retinas as controls and fibrovascular membranes (FVMs) in patients with PDR as PDR samples. The functional term and signaling pathway enrichment analysis of the selected genes were subsequently performed. In addition, protein-protein interaction (PPI) networks were constructed to determine the hub genes, and the network of transcriptional factor (TF) and target hub genes was also analyzed. In total, 132 hypomethylated genes were found to be upregulated, whereas 172 hypermethylated genes were discovered to be downregulated in PDR. The hypomethylated upregulated genes were found to be enriched in the pathways, such as "cell-substrate adhesion", "adherens junction", "cell adhesion molecule binding" and "extracellular matrix receptor interactions". Meanwhile, the hypermethylated downregulated genes were enriched in the pathways, such as "visual perception", "presynapse" and the "synaptic vesicle cycle". Based on the PPI analysis, a total of eight hub genes were identified: CTGF, SERPINH1, LOX, RBP3, OTX2, RPE65, OPN1SW and NRL. It was hypothesized that the aberrant methylation of these genes might be related to the possible pathophysiology of PDR. An important transcriptional factor, TFDP1, was discovered to share the closest interactions with the hub genes from the gene-TF network. In conclusion, the present study identified an association among DNA methylation and gene expression in PDR using bioinformatics analysis, and identified the hub genes which might be potential methylation-based diagnosis and treatment targets for PDR in the near future.
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Retinopatía Diabética/genética , Proteínas del Ojo/genética , Regulación de la Expresión Génica , Metilación de ADN , Retinopatía Diabética/metabolismo , Proteínas del Ojo/metabolismo , Perfilación de la Expresión Génica , Redes Reguladoras de Genes , Humanos , Transducción de Señal/genéticaRESUMEN
OBJECTIVES: Aim was to evaluate endocrine, metabolic and thyroid parameters which could help to explore the relationship between thyroid function and metabolic changes in Chinese polycystic ovary syndrome (PCOS) patients. METHODS: Within a prospective study in PCOS patients compared with healthy women, thyroid parameters were assessed, as well as changes of endocrine, metabolic and clinical characteristics. RESULTS: 144 PCOS patients and 48 normo-ovulatory women matched by age as controls were included. There were significant increases of thyroid-stimulating hormone (TSH), total triiodothyronine (TT3) and total thyroxine (TT4) in the PCOS patients. Body mass index (BMI), waist and hip ratio (WHR), luteinizing hormone (LH), LH/follicle-stimulating hormone (FSH) and total testosterone (T) were significantly higher in PCOS compared with the controls. Total cholesterol (CHO), triglycerides (TG) and apolipoprotein B (ApoB) levels in PCOS were higher, whereas high-density lipoprotein (HDL) and apolipoprotein A (ApoA) were lower compared with the controls. Insulin and homeostatic model assessment of insulin resistance (HOMA-IR) were significantly higher in patients with PCOS. CONCLUSIONS: Our study confirms the well-known negative metabolic changes in PCOS patients. The small increases of TSH, TT3 and TT4 level may be related with these metabolic changes in PCOS patients. Further studies may improve the understanding of the relationship between thyroid function and metabolic changes.
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Resistencia a la Insulina/fisiología , Síndrome del Ovario Poliquístico/sangre , Tirotropina/sangre , Tiroxina/sangre , Triyodotironina/sangre , Adulto , Apolipoproteínas A/sangre , Apolipoproteínas B/sangre , Índice de Masa Corporal , China , Colesterol/sangre , Femenino , Hormona Folículo Estimulante/sangre , Humanos , Lipoproteínas HDL/sangre , Hormona Luteinizante/sangre , Estudios Prospectivos , Testosterona/sangre , Relación Cintura-Cadera , Adulto JovenRESUMEN
Background: Uncontrolled inflammation is a typical feature of sepsis-related lung injury. The key event in the progression of lung injury is Caspase-1-dependent alveolar macrophage (AM) pyroptosis. Similarly, neutrophils are stimulated to release neutrophil extracellular traps (NETs) to participate in the innate immune response. This study aims to illustrate the specific mechanisms by which NETs activate AM at the post-translational level and maintain lung inflammation. Methods: We established a septic lung injury model by caecal ligation and puncture. We found elevated NETs and interleukin-1b (IL-1ß) levels in the lung tissues of septic mice. Western blot and immunofluorescence analyses was utilized to determine whether NETs promote AM pyroptosis and whether degrading NETs or targeting the NLRP3 inflammasome had protective effects on AM pyroptosis and lung injury. Flow cytometric and co-immunoprecipitation analyses verified intracellular reactive oxygen species (ROS) levels and the binding of NLRP3 and ubiquitin (UB) molecules, respectively. Results: Increased NETs production and IL-1ß release in septic mice were correlated with the degree of lung injury. NETs upregulated the level of NLRP3, followed by NLRP3 inflammasome assembly and caspase-1 activation, leading to AM pyroptosis executed by the activated fragment of full-length gasdermin D (FH-GSDMD). However, the opposite effect was observed in the context of NETs degradation. Furthermore, NETs markedly elicited an increase in ROS, which facilitated the activation of NLRP3 deubiquitination and the subsequent pyroptosis pathway in AM. Removal of ROS could promote the binding of NLRP3 and ubiquitin, inhibit NLRP3 binding to apoptosis-associated spotted proteins (ASC) and further alleviate the inflammatory changes in the lungs. Conclusion: In summary, these findings indicate that NETs prime ROS generation, which promotes NLRP3 inflammasome activation at the post-translational level to mediate AM pyroptosis and sustain lung injury in septic mice.
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Purpose: To demonstrate an interaction-based method for the refinement of Gene Set Enrichment Analysis (GSEA) results. Method: Intravitreal injection of miR-124-3p antagomir was used to knockdown the expression of miR-124-3p in mouse retina at postnatal day 3 (P3). Whole retinal RNA was extracted for mRNA transcriptome sequencing at P9. After preprocessing the dataset, GSEA was performed, and the leading-edge subsets were obtained. The Apriori algorithm was used to identify the frequent genes or gene sets from the union of the leading-edge subsets. A new statistic d was introduced to evaluate the frequent genes or gene sets. Reverse transcription quantitative PCR (RT-qPCR) was performed to validate the expression trend of candidate genes after the knockdown of miR-124-3p. Results: A total of 115,140 assembled transcript sequences were obtained from the clean data. With GSEA, the NOD-like receptor signaling pathway, C-type-like lectin receptor signaling pathway, phagosome, necroptosis, JAK-STAT signaling pathway, Toll-like receptor signaling pathway, leukocyte transendothelial migration, chemokine signaling pathway, NF-kappa B signaling pathway and RIG-I-like signaling pathway were identified as the top 10 enriched pathways, and their leading-edge subsets were obtained. After being refined by the Apriori algorithm and sorted by the value of the modulus of d , Prkcd, Irf9, Stat3, Cxcl12, Stat1, Stat2, Isg15, Eif2ak2, Il6st, Pdgfra, Socs4 and Csf2ra had the significant number of interactions and the greatest value of d to downstream genes among all frequent transactions. Results of RT-qPCR validation for the expression of candidate genes after the knockdown of miR-124-3p showed a similar trend to the RNA-Seq results. Conclusion: This study indicated that using the Apriori algorithm and defining the statistic d was a novel way to refine the GSEA results. We hope to convey the intricacies from the computational results to the low-throughput experiments, and to plan experimental investigations specifically.
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Stem cell replacement therapy has emerged as one of the most promising treatment options for retinal degenerative diseases, which are the main causes of irreversible vision loss. Three-dimensional (3D) retinal organoid culture is a cutting-edge technology for differentiating embryonic stem cells into retinal cells by forming a laminated retinal structure. However, 3D culture systems have strict requirements with respect to the experimental environment and culture technologies. Our study aimed to investigate the effect of retinal conditioned medium (RCM) at different developmental stages on the early differentiation of embryonic stem cells into retina in a 3D culture system. In this study, we added RCM to the 3D culture system and found that it could promote the differentiation of mouse embryonic stem cells (mESCs) into neuroretina. We further explored the possible mechanisms of RCM that regulate differentiation through proteomic analysis. RCM at different time points disclosed different protein profiles. Proteins which improved energy metabolism of mESCs might help improve the viability of embryonic bodies. We then screened out Snap25, Cntn1, Negr1, Dpysl2, Dpysl3, and Crmp1 as candidate proteins that might play roles in the differentiation and neurogenesis processes of mESCs, hoping to provide a basis for optimizing a retinal differentiation protocol from embryonic stem cells.
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Células Madre Embrionarias , Proteómica , Animales , Ratones , Medios de Cultivo Condicionados/farmacología , Medios de Cultivo Condicionados/metabolismo , Diferenciación Celular , Retina/metabolismoRESUMEN
Purpose: The purpose of this study was to investigate the effects and mechanism of microRNA (miR)-92a-3p in retinal angiogenesis in vitro and in vivo. Methods: The expression of miR-92a-3p was verified by real-time quantitative polymerase chain reaction (RT-qPCR). Agomir-92a-3p was intravitreally injected into the right eye on postnatal day 3 (P3), P5, and P8 in the mice, with the agomir-NC injected left eye as the control. At P7, P9, and P12, immunofluorescence was performed to examine the retinal superficial vascular plexus, deep vascular plexus, proliferation, and apoptosis in retinal vascular endothelial cells (ECs). Human retinal microvascular endothelial cells (HRMECs) were treated with mimic-NC and mimic-92a-3p, then the tube formation, cell migration, and wound healing assays were used to detect the effect of miR-92a-3p on retinal angiogenesis in vitro. Agomir-92a-3p was also intravitreally injected into the right eye of oxygen-induced retinopathy (OIR) mice at P12, with the agomir-NC injected left eye as the control, the neovascularization was observed by retinal flatmount staining with isolectin B4 at P17. Bioinformatics and high-throughput sequencing were performed to identify potential target genes of miR-92a-3p. RT-qPCR and Western blot were carried out to detect the expression of SGK3, p-GSK3ß, GSK3ß, Bcl-xL, and cleaved caspase-3 in the HRMECs and mouse retinas. Results: The overexpression of miR-92a-3p inhibited the development of retinal superficial vascular plexus and deep vascular plexus, decreased the expression of Ki67, and increased the expression of cleaved caspase-3 in isolectin B4-labeled retinal vascular ECs. In vitro, the overexpression of miR-92a-3p markedly suppressed the tube formation, cell migration, and wound healing of cultured ECs. Overexpression of miR-92a-3p inhibited both in vivo and in vitro physiological angiogenesis by downregulating the expression of SGK3, p-GSK3ß/GSK3ß, and Bcl-xL. In addition, agomir-92a-3p inhibited the pathological retinal neovascularization of OIR mice, by targeting SGK3, p-GSK3ß/GSK3ß, and Bcl-xL. Conclusions: The miR-92a-3p could affect retinal angiogenesis by targeting SGK3 pathway, suggesting that miR-92a-3p may be a potential anti-angiogenic factor for retinal vascular disease.
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MicroARNs , Neovascularización Patológica , Proteínas Serina-Treonina Quinasas , Retina , Animales , Humanos , Ratones , Caspasa 3/metabolismo , Proliferación Celular/genética , Células Endoteliales/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Antígeno Ki-67/metabolismo , Lectinas , MicroARNs/genética , MicroARNs/metabolismo , Neovascularización Patológica/genética , Neovascularización Patológica/metabolismo , Oxígeno/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Retina/patologíaRESUMEN
Acute lung injury (ALI) is a common lung pathology that is accompanied by alveolar macrophage (AM) activation and inflammatory response. This study investigated the role of the long non-coding RNA NONRATT004344 (hereafter named lncRNA NLRP3) in regulating the Nod-like receptor protein 3 (NLRP3)-triggered inflammatory response in early ALI and the underlying mechanism as well. We established LPS-induced ALI models to explore their interactive mechanisms in vitro and in vivo. Luciferase reporter assays were performed to determine that miR-138-5p could bind to lncRNA NLRP3 and NLRP3. We observed increased lncRNA NLRP3 expression, decreased miR-138-5p expression, NLRP3 inflammasome activation, and upregulated caspase-1, IL-1ß, and IL-18 expression in the LPS-induced ALI model. Furthermore, lncRNA NLRP3 overexpression activated the NLRP3 inflammasome and promoted IL-1ß and IL-18 secretion; the miR-138-5p mimic abolished these effects in vivo and in vitro. Consistently, miR-138-5p inhibition reversed the effects of lncRNA NLRP3 silencing on the expression of NLRP3-related molecules and inhibition of the NLRP3/caspase-1/IL-1ß signalling pathway. Mechanistically, lncRNA NLRP3 sponging miR-138-5p facilitated NLRP3 activation through a competitive endogenous RNA (ceRNA) mechanism. In summary, our results suggested that lncRNA NLRP3 binding miR-138-5p promotes NLRP3-triggered inflammatory response via lncRNA NLRP3/miR-138-5p/NLRP3 ceRNA network (ceRNET) and provides insights into the treatment of early ALI.
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Lesión Pulmonar Aguda/genética , Inflamación/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , ARN Largo no Codificante/metabolismo , Regiones no Traducidas 3'/genética , Lesión Pulmonar Aguda/patología , Animales , Secuencia de Bases , Caspasa 1/metabolismo , Línea Celular , Inflamasomas/metabolismo , Inflamación/patología , Interleucina-18/metabolismo , Interleucina-1beta/metabolismo , Lipopolisacáridos , Pulmón/metabolismo , Pulmón/patología , Masculino , MicroARNs/genética , MicroARNs/metabolismo , Modelos Biológicos , ARN Largo no Codificante/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Sprague-Dawley , Transcripción Genética , Regulación hacia Arriba/genéticaRESUMEN
OBJECTIVE: To investigate the dependency of menopausal symptoms on age and/or menopausal status and association with social and environmental factors. METHODS: The cross-sectional study was conducted on 4595 women (40-83 years) coming from 31 provinces during two years to our "Menopause Clinic", the first official center in China. Menopausal symptoms were assessed: negative mood, cognitive symptoms, sleep disorder, vasomotor symptoms (VMS), urogenital symptoms, autonomic nervous disorder, limb pain/paresthesia. Social and environmental factors were collected; simple and unconditional logistic regression with adjustments by all analyzed factors were used to assess associations. RESULTS: Urogenital symptoms were the most common and VMS the least common complaints. All symptoms, except cognitive and urogenital symptoms, worsened age-dependently up to 60 years but improved beyond this age. Most symptoms also were associated with menopause, except negative mood and autonomic nervous disorders. Soya-rich diet decreased all symptoms, but only if consumed daily. Exercise was beneficial for some symptoms. Hormone replacement therapy (HRT) was most effective but only with regular use. Increased alcohol consumption aggravated VMS. Higher education was associated with less symptoms; no relationship was found for smoking, gravidity, parity, and menarche. CONCLUSION: All symptoms, except cognitive and urogenital symptoms, worsened age-dependently up to 60 years but improved beyond this age; most were also associated with menopause. For the first time in a large study population, it was observed that soy-rich diet is protective but only with daily consumption. Exercising can protect against some of the symptoms. HRT decreased all symptoms, but regular use is necessary. Women with higher education reported less symptoms, but after adjustments no other relationships were observed (ChiCTR2000035047).
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Estado de Salud , Menopausia/psicología , Calidad de Vida , Ansiedad/etiología , Pueblo Asiatico , China/epidemiología , Estudios Transversales , Dieta/estadística & datos numéricos , Femenino , Humanos , Modelos Logísticos , Menopausia/fisiología , Persona de Mediana Edad , Trastornos del Sueño-Vigilia/etiología , Factores SocioeconómicosRESUMEN
PURPOSE: To study the quality of our human ovarian tissue cryopreservation technique as performed in the first official "International Fertility Protection Centre" in China in patients with certain cancer types using a mouse model, and to find the best site for tissue transplantation in the mouse. METHODS: Thirty-six BALB/C female nude mice were randomly divided into 3 groups, group 1: control group; group 2: ovariectomized group; group 3: ovarian tissue transplantation group. Seventy-two pieces obtained from six ovarian tissue samples from each of three cancer patients were transplanted into the ovarian bursa cavity (OBC), the subcutaneous thigh (TS) and the subcutaneous neck (NS) and removed after 1.5 and 2.5 months, respectively. Follicular growth rate (FGR), total follicle surviving rate (TFSR), tissue recovery rate (TRR), antral follicles (AF), follicle stimulating hormone (FSH), estradiol (E2) and anti-Mullerian hormone (AMH) levels were measured. RESULTS: No significant differences in FGR, OBC, NS (p > 0.05); TFSR was 100% in OBC, NS and TS. No significant differences in TRR (p > 0.05); AF were found only in OBC; TFSR was 100% after transplantation; significantly higher FGR in the 2.5 months compared to the 1.5 months-group (p < 0.05). AMH- and E2-level in group 1 and 3 were significantly higher than in group 2 (p < 0.05); in contrast, FSH was significantly lower. CONCLUSIONS: After transplantation in the mice, the thawed ovarian tissue survived and follicles developed. The ovarian fossa site was the best site for transplantation. Our animal experiments can verify that our human ovarian tissue cryopreservation technique can preserve the quality of ovarian tissue. This is the essential precondition for successful re-transplantation into the patients after performing chemo/radiotherapy to protect ovarian function and fertility.
Asunto(s)
Criopreservación , Xenoinjertos/fisiología , Folículo Ovárico/fisiología , Folículo Ovárico/trasplante , Ovario , Adulto , Animales , Hormona Antimülleriana/metabolismo , Estradiol/metabolismo , Femenino , Preservación de la Fertilidad , Hormona Folículo Estimulante/metabolismo , Supervivencia de Injerto , Xenoinjertos/crecimiento & desarrollo , Xenoinjertos/metabolismo , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/metabolismo , Ovario/cirugía , Técnicas de Cultivo de Tejidos , Trasplante Heterólogo , Neoplasias del Cuello Uterino/cirugíaRESUMEN
INTRODUCTION: The climacteric symptoms during peri- and postmenopause have not been extensively studied in China. To further understanding of the characteristics of Chinese women during their menopausal transition, the aim of the study was to compare the prevalence and severity of climacteric symptoms of peri- and postmenopausal Chinese women. METHODS: The retrospective descriptive study was carried out in perimenopausal and postmenopausal women from 20 provinces of China who visited Beijing Obstetrics & Gynecology hospital during 2008-2015. A total of 1225 women aged 34-76 years without hormone replacement therapy were analyzed. Menopausal status was defined by the "2011 Stages of Reproductive Aging Workshop criteria". The following climacteric symptoms were assessed: fatigue, insomnia, irritability, depression, palpitations, muscle and joint pain, dizziness, vaginal dryness, headache, urinary incontinence, hot flash, sweat, pain during intercourse, and formication. RESULTS: The most frequent five symptoms were fatigue (75.84%), insomnia (69.39%), irritability (67.02%), palpitations (62.78%), and depression (61.88%). The prevalence of 14 symptoms was significantly higher in the postmenopause compared with the perimenopause status group (all p < .05).The severity of fatigue, insomnia, irritability, palpitation, vaginal dryness, muscle, and joint pain and pain during intercourse was significantly different between the perimenopausal and postmenopausal groups. CONCLUSIONS: The most frequent five symptoms among the investigated 1225 Chinese women were fatigue, insomnia, irritability, palpitations, depression, nearly the same in perimenopausal and postmenopausal women. The prevalence and the severity of most of the symptoms were significantly different between the two groups.