Assuntos
Estado Terminal , Rim , Humanos , Incidência , Testes de Função Renal , Fatores de Risco , Antibacterianos , CreatininaRESUMO
Ferroptosis is a recently identified form of cell death characterized by iron-dependent lipid peroxidation. Understanding the effects of lipid peroxidation on cellular processes during ferroptosis requires insights into lipid droplets (LDs) and their viscosity changes. To gain further insights into the intricacies of ferroptosis, it is crucial to have a fluorescent probe that targets LDs and responds to changes in viscosity. In this study, we introduce a novel LD-targeting viscosity fluorescent probe named TQE, based on the principles of aggregation-induced emission (AIE). The probe displayed AIE characteristics in tetrahydrofuran, possessing a partition coefficient (logP) of 5.87. With increased viscosity, intramolecular rotation was restricted, leading to a remarkable 3.3-fold enhancement in emission. Notably, TQE exhibited robust resistance to photo-bleaching during cellular imaging, maintaining approximately 75% of its emission intensity even after 30 min of laser irradiation. Importantly, the AIEgen could not generate hydroxyl radicals when exposed to light for up to 3 h, suggesting the low photo-toxicity of TQE to cells. Leveraging these properties, we successfully employed the probe for fluorescent imaging of the viscosity change in LDs during ferroptosis.
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miRNAs are a family of short, noncoding RNAs that are involved in many processes in melanoma cells. MITF acts as a master regulator of melanocyte function, development and survival by modulating various genes. Hydroxyurea (HU) is used to treat melanoma, and miRNA expression is altered after HU treatment in B16 melanoma cells. In this study, we screened for miRNAs that were upregulated after HU treatment and that targeted the MITF gene. We found that miR-7013-3p exhibited increased expression after HU treatment and could bind to MITF. miR-7013-3p inhibited melanin production, proliferation, and migration and promoted apoptosis in B16 melanoma cells. The results may provide more information on the roles of miR-7013-3p in B16 melanoma cells.
Assuntos
Hidroxiureia/farmacologia , Melanoma/tratamento farmacológico , MicroRNAs/metabolismo , Fator de Transcrição Associado à Microftalmia/genética , Neoplasias Cutâneas/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Hidroxiureia/uso terapêutico , Melaninas/biossíntese , Melanoma/genética , Melanoma/patologia , Camundongos , MicroRNAs/genética , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/patologia , Regulação para Cima/efeitos dos fármacosRESUMO
A range of preliminary designs of customised total knee implants (CTKIs) was created by resurfacing the distal femur and applying different tibial bearing surface curvatures. These were then compared with a scaled off-the-shelf symmetric total knee implant (STKI). To evaluate the biomechanical performance, a dynamic knee simulation model was created with patient-specific muscle and ankle joint loads calculated from an OpenSim musculoskeletal model. Simulation results showed the transverse curvatures of the tibial bearing surface influenced femoral mediolateral translation, while its longitudinal curvatures affected femoral adduction. Compared to the STKI, the CTKIs could restore patient knee function.
Assuntos
Artroplastia do Joelho , Prótese do Joelho , Movimento (Física) , Postura/fisiologia , Tíbia/fisiologia , Tíbia/cirurgia , Fenômenos Biomecânicos , Força Compressiva , Simulação por Computador , Fêmur/diagnóstico por imagem , Análise de Elementos Finitos , Humanos , Articulação do Joelho/diagnóstico por imagem , Ligamentos/diagnóstico por imagem , Amplitude de Movimento Articular/fisiologia , Estresse Mecânico , Tíbia/diagnóstico por imagem , Suporte de CargaRESUMO
The fat content of milk determines the quality of milk, and triglycerides are the major components of milk fat. Milk fat synthesis is regulated by many factors. Lipopolysaccharide (LPS) has been shown to inhibit milk fat synthesis in bovine mammary epithelial cells, but research on the underlying mechanisms has been limited. MicroRNA (miRNA) are involved in many physiological processes, but there have been few studies on their regulation in milk fat synthesis. In this study, we aimed to investigate whether LPS upregulates miR-27a-3p, which targets PPARG, thereby inhibiting the synthesis of triglycerides in a dairy cow mammary epithelial cell line (MAC-T). After LPS stimulation of MAC-T cells, PPARG gene expression and milk fat synthesis were inhibited. TargetScan software was used to predict miRNA targeting PPARG, and miR-27a-3p was selected as a candidate. A dual luciferase reporter assay further confirmed the targeting connection between miR-27a-3p and the PPARG gene. To investigate the functions of miR-27a-3p, miR-27a-3p mimic and inhibitors were transfected into MAC-T cells. The mRNA and protein levels of PPAR-γ were negatively correlated with the expression of miR-27a-3p. Lipid droplet accumulation and triglyceride synthesis were also negatively correlated with miR-27a-3p expression. Inhibition of miR-27a-3p partially reversed the LPS-induced decreases in PPARG expression and milk fat synthesis. In summary, our results reveal that LPS can inhibit MAC-T cell milk fat synthesis by upregulating miR-27a-3p, which targets the PPARG gene.
Assuntos
Lipopolissacarídeos/farmacologia , Glândulas Mamárias Animais/metabolismo , MicroRNAs/metabolismo , PPAR gama/genética , Triglicerídeos/biossíntese , Animais , Bovinos , Contagem de Células/veterinária , Linhagem Celular , Células Epiteliais/metabolismo , Feminino , Leite/citologia , PPAR gama/metabolismo , RNA Mensageiro/metabolismo , Ativação Transcricional , Regulação para CimaRESUMO
The pituitary gland functions as a prominent regulator of diverse physiologic processes by secreting multiple hormones. Circular RNAs (circRNAs) are an emerging novel type of endogenous noncoding RNA that have recently been recognized as powerful regulators participating in various biological processes. However, the physiological roles and molecular mechanisms of circRNAs in pituitary remain largely unclear. Herein, we concentrated on expounding the biological function and molecular mechanism of circRNA in rat pituitary. In this study, we identified a novel circRNA in pituitary tissue, circAkap17b, which was pituitary- and stage-specific. Then, we designed circAkap17b siRNA and constructed an overexpression plasmid to evaluate the effect of loss- and gain-of-circAkap17b function on FSH secretion. Interestingly, silencing circAkakp17b significantly inhibited FSH expression and secretion, while overexpression of circAkap17b enhanced FSH expression and secretion. Furthermore, dual luciferase reporter and RNA immunoprecipitation (RIP) assays confirmed that circAkap17b could serve as miR-7 sponge to regulate target genes. Additionally, miR-7b suppressed FSH expression and secretion by directly targeting Fshb through the dual luciferase reporter and RT-qPCR analysis. Additionally, rescue experiments showed that circAkap17b could regulate FSH secretion in pituitary cells through a circAkap17b-miR-7-Fshb axis. Collectively, we demonstrated that circAkap17b could act as a molecular sponge of miR-7 to upregulate expression of the target gene Fshb and facilitate FSH secretion. These findings provide evidence for a novel regulatory role of circRNAs in pituitary.
Assuntos
Hormônio Foliculoestimulante/biossíntese , Hormônio Foliculoestimulante/genética , Regulação da Expressão Gênica , MicroRNAs/genética , Hipófise/metabolismo , RNA Circular/genética , Animais , Células Cultivadas , Hipófise/citologia , Ratos , Reprodução/genéticaRESUMO
BACKGROUND: Matrix metalloproteinases (MMPs) are a family of zinc endopeptidases that play a key role in both physiological and pathological tissue degradation. MMPs have reportedly shown great potentials in the degradation of the Extracellular Matrix (ECM), have shown great potentials in targeting bioactive and imaging agents in cancer treatment. MMPs could provoke Epithelial to Mesenchymal Transition (EMT) of cancer cells and manipulate their signaling, adhesion, migration and invasion to promote cancer cell aggressiveness. Therefore, targeting and particularly inhibiting MMPs within the tumor microenvironment is an effective strategy for cancer treatment. Based on this idea, different MMP inhibitors (MMPIs) have been developed to manipulate the tumor microenvironment towards conditions appropriate for the actions of antitumor agents. Studies are ongoing to improve the selectivity and specificity of MMPIs. Structural optimization has facilitated the discovery of selective inhibitors of the MMPs. However, so far no selective inhibitor for MMP-7 has been proposed. AIMS: This study aims to comprehensively review the potentials and advances in applications of MMPs particularly MMP-7 in targeted cancer treatment approaches with the main focus on targeted drug delivery. Different targeting strategies for manipulating and inhibiting MMPs for the treatment of cancer are discussed. MMPs are upregulated at all stages of expression in cancers. Different MMP subtypes have shown significant targeting applicability at the genetic, protein, and activity levels in both physiological and pathophysiological conditions in a variety of cancers. The expression of MMPs significantly increases at advanced cancer stages, which can be used for controlled release in cancers in advance stages. METHODS: Moreover, this study presents the synthesis and characteristics of a new and highly selective inhibitor against MMP-7 and discusses its applications in targeted drug delivery systems for therapeutics and diagnostics modalities. RESULTS: Our findings showed that the structure of the inhibitor P3' side chains play the crucial role in developing an optimized MMP-7 inhibitor with high selectivity and significant degradation activities against ECM. CONCLUSION: Optimized NDC can serve as a highly potent and selective inhibitor against MMP-7 following screening and optimization of the P3' side chains, with a Ki of 38.6 nM and an inhibitory selectivity of 575 of MMP-7 over MMP-1.
Assuntos
Antineoplásicos/farmacologia , Sistemas de Liberação de Medicamentos , Metaloproteinase 7 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/farmacologia , Neoplasias/tratamento farmacológico , Antineoplásicos/síntese química , Antineoplásicos/química , Portadores de Fármacos/química , Humanos , Inibidores de Metaloproteinases de Matriz/síntese química , Inibidores de Metaloproteinases de Matriz/química , Nanoestruturas/química , Neoplasias/diagnóstico , Neoplasias/metabolismoRESUMO
Few studies have been conducted to investigate kinematics and kinetics of the patellofemoral joint under physiological muscle forces and ankle joint loads. In this study, a preliminary design of a customised total knee implant was proposed and created. To compare the influences of different patella treatment scenarios, a dynamic knee simulation model was created with patient-specific muscle forces and ankle joint loads that are calculated from an OpenSim musculoskeletal model. The goal is to improve patellar implant-bone connection and restore patellofemoral joint mobility. Identical dynamic boundary conditions were applied on an unresurfaced patella and three different dome-shaped patellar implants. It was found that the unresurfaced patella and patellar implants resulted in different motions of patellar internal rotation and medial tilt. The size of the dome-shaped patellar implant affected the motion and loading of the patellofemoral joint. When the exposed patella bone was not fully covered by the patellar implant, the patella bone then contacted the femoral component during knee flexion. This would most likely lead to anterior knee pain and subsequent revision.
Assuntos
Artroplastia do Joelho , Prótese do Joelho , Articulação Patelofemoral , Fenômenos Biomecânicos , Humanos , Articulação do Joelho/cirurgia , Patela/cirurgia , Articulação Patelofemoral/cirurgia , Amplitude de Movimento Articular , RotaçãoRESUMO
Growth hormone (GH) is an important hormone released by the pituitary gland that plays a key role in the growth and development of organisms. In our study, TargetScan analysis and the dual luciferase reporter assays were used to predict and screen for miRNAs that might act on the rat Gh1 gene, and we identified miR-543-5p. Then, the GH3 cell line and the primary rat pituitary cells were transfected with miRNA mimic, inhibitor, and siRNA. We detected the Gh1 gene expression and the GH secretion by real-time PCR and ELISAs, respectively, to verify the regulatory effect of miR-543-5p on GH secretion. The results showed that miR-543-5p can inhibit Gh1 mRNA expression and reduce GH secretion. MiR-543-5p inhibitor upregulated Gh1 mRNA expression and increased GH secretion compared with the negative control. In summary, miR-543-5p downregulates Gh1 expression, resulting in a decrease in GH synthesis and secretion, which demonstrates the important role of miRNAs in regulating GH and animal growth and development.
Assuntos
Hormônio do Crescimento/genética , MicroRNAs/genética , Hormônios Adeno-Hipofisários/genética , Regiões 3' não Traduzidas/genética , Animais , Linhagem Celular , Expressão Gênica , Regulação da Expressão Gênica/genética , Hormônio do Crescimento/metabolismo , Masculino , Hipófise/metabolismo , Adeno-Hipófise/metabolismo , Hormônios Adeno-Hipofisários/metabolismo , Cultura Primária de Células , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , TransfecçãoRESUMO
Customised total knee replacement could be the future therapy for knee joint osteoarthritis. A preliminary design of a customised total knee implant based on knee anatomy was studied in this article. To evaluate its biomechanical performance, a dynamic finite element model based on the Oxford knee rig was created to simulate a squatting motion. Unlike previous research, this dynamic model was simulated with patient-specific muscle and joint loads that were calculated from an OpenSim musculoskeletal model. The dynamic response of the customised total knee implant was simulated under three cruciate ligament scenarios: both cruciate ligaments retained, only anterior cruciate ligament removed and both cruciate ligaments removed. In addition, an off-the-shelf symmetric total knee implant with retained cruciate ligaments was simulated for comparison analysis. The customised total knee implant with both cruciate ligaments retained showed larger ranges of femoral external rotation and posterior translation than the symmetric total knee implant. The motion of the customised total knee implant was also in good agreement with a healthy knee. There were no big differences in the tibiofemoral compressive forces in the customised total knee implant model under the three scenarios. These forces were generally consistent with other experimental and simulation results. However, the customised total knee implant design resulted in larger tibiofemoral compressive force than the symmetric total knee implant after 50° knee flexion, which was caused by the larger tibiofemoral relative motion.
Assuntos
Análise de Elementos Finitos , Articulação do Joelho/fisiologia , Articulação do Joelho/cirurgia , Prótese do Joelho , Fenômenos Mecânicos , Músculos/fisiologia , Postura , Fenômenos Biomecânicos , HumanosRESUMO
Long noncoding RNAs (lncRNAs) are important regulators that have multiple functions in a variety of biological processes. However, the contributions of lncRNAs to follicle-stimulating hormone (FSH) secretion remain largely unknown. In this study, we first identified a novel lncRNA, lncRNA-m433s1, as an intergenic lncRNA located in the cytoplasm. We next used MS2-RIP assays to demonstrate that lncRNA-m433s1 interacted with miR-433. Furthermore, we detected the levels of lncRNA-m433s1, miR-433, and Fshß expression, FSH concentrations, and apoptosis upon overexpression and knockdown of lncRNA-m433s1, revealing that lncRNA-m433s1 upregulated Fshß expression. Globally, lncRNA-m433s1 reduced the inhibitory effect of miR-433 on Fshß and further regulated FSH secretion as a competing endogenous RNA (ceRNA) by sponging miR-433. This ceRNA model will provide novel insight into the regulatory mechanisms of lncRNAs associated with rat reproduction.
Assuntos
MicroRNAs/metabolismo , Adeno-Hipófise/citologia , Animais , Feminino , Subunidade beta do Hormônio Folículoestimulante/genética , Subunidade beta do Hormônio Folículoestimulante/metabolismo , Regulação da Expressão Gênica/fisiologia , Masculino , MicroRNAs/genética , RNA Longo não Codificante/genética , Ratos , Ratos Sprague-Dawley , Regulação para CimaRESUMO
The follicle-stimulating hormone (FSH), which is synthesized and secreted by the anterior pituitary gland, plays an important role in regulating reproductive processes. In this study, using the TargetScan program, we predicted that microRNAs (miRNAs) regulate FSH secretion. Dual-luciferase reporter assays were performed and identified miR-7a-5p. MiR-7a-5p has been reported to regulate diverse cellular functions. However, it is unclear whether miR-7a-5p binds to mRNAs and regulates reproductive functions. Therefore, we constructed a suspension of rat anterior pituitary cells and cultured them under adaptive conditions, transfected miR-7a-5p mimics or inhibitor into the cell suspension and detected expression of the FSHb gene. The results demonstrated that miR-7a-5p downregulated FSHb expression levels, while treatment with miR-7a-5p inhibitors upregulated FSHb expression levels relative to those of negative control groups, as shown by quantitative PCR analysis. The results were confirmed with a subsequent experiment showing that FSH secretion was reduced after treatment with mimics and increased in the inhibitor groups, as shown by enzyme-linked immunosorbent assay. Our results indicated that miR-7a-5p downregulates FSHb expression levels, resulting in decreased FSH synthesis and secretion, which demonstrates the important role of miRNAs in the regulation of FSH and animal reproduction.
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Circular RNAs (circRNAs) are a new class of RNA that have a stable structure characterized by covalently closed circular molecules and are involved in invasive pituitary adenomas and recurrent clinically nonfunctioning pituitary adenomas. However, information on circRNAs in the normal pituitary, especially in rats, is limited. In this study, we identified 4123 circRNAs in the immature (D15) and mature (D120) rat anterior pituitary using the Illumina platform, and 32 differentially expressed circRNAs were found. A total of 150 Gene Ontology terms were significantly enriched, and 16 KEGG pathways were found to contain differentially expressed genes. Moreover, we randomly selected eight highly expressed circRNAs and detected their relative expression levels in the mature and immature rat pituitary by qPCR. In addition, we predicted 90 interactions between 53 circRNAs and 57 miRNAs using miRanda. Notably, circ_0000964 and circ_0001303 are potential miRNA sponges that may regulate the Fshb gene. The expression profile of circRNAs in the immature and mature rat anterior pituitary may provide more information about the roles of circRNAs in the development and reproduction in mammals.
Assuntos
Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/genética , Adeno-Hipófise/metabolismo , RNA Circular/genética , Fatores Etários , Animais , Análise por Conglomerados , Redes Reguladoras de Genes , MicroRNAs/metabolismo , Adeno-Hipófise/crescimento & desenvolvimento , RNA Circular/metabolismo , Ratos Sprague-DawleyRESUMO
miRNAs have been shown to regulate a variety of biological process. It has been shown that miR-181a regulates porcine adipogenesis by targeting Tumor Necrosis Factor-α (TNF-α), but the overall functions of miR-181a in porcine preadipocyte differentiation remain unclear. This study aimed to explore the functions of miR-181a in porcine preadipocyte differentiation via the TGFß/Smad pathway. The TargetScan program was used to predict miRNAs targeting TGFBR1, and miR-181a was selected as a candidate. To investigate the functions of miR-181a, miRNA mimics and inhibitors were used to overexpress or knockdown miR-181a, respectively. RT-qPCR and Western blotting were used to measure the expression of aP2, PPARγ, C/EBPα and TGFBR1 in porcine preadipocytes. Lipid accumulation and adipocyte apoptosis were detected using Oil Red O staining and flow cytometry, respectively. Taken together, our results indicated that miR-181a promoted porcine preadipocyte differentiation by directly targeting TGFBR1.
Assuntos
Adipócitos/fisiologia , Adipogenia/genética , Diferenciação Celular/genética , MicroRNAs/fisiologia , Proteínas Serina-Treonina Quinases/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Animais , Células Cultivadas , Regulação da Expressão Gênica , MicroRNAs/genética , Receptor do Fator de Crescimento Transformador beta Tipo I , SuínosRESUMO
To evaluate the clinical efficacy and safety of tripterygium glycosides (TG) in the treatment of henoch-schonlein purpura nephritis(HSPN). Seven English and Chinese databases (up to Nov. 9, 2017), were searched to collect the RCTs on TG for HSPN. Two researchers independently screened the literature according to inclusion criteria and exclusion criteria, extracted data, and evaluated the quality of the literature. After completion, cross-checking was performed and Meta-analysis was performed using RevMan 5.3 software. At the same time, different outcomes of the interventions were analyzed subgroupically. A total of 46 RCTs were included, with 1 659 in the experimental group and 1 596 in the control group. All the clinical studies showed a low quality. In terms of complete remission rate, the group with TG performed better than the group with conventional therapy or GCï¼RR=1.82ï¼95%CI[1.39ï¼2.39];RR=2.03ï¼95%CI[1.37ï¼2.99]ï¼ï¼the group with TG+GC performed better than the group with GCï¼RR=1.46ï¼95%CI[1.32ï¼1.60]ï¼ï¼and the group with CTX+GC performed better than the group with TG+GCï¼RR=0.35ï¼95%CI[0.16ï¼0.75]ï¼. In terms of total effective rate, the group with TG performed better than the group with conventional therapy or GCï¼RR=1.44ï¼95%CI[1.19,1.74];RR=1.30ï¼95%CI[1.16ï¼1.46]ï¼ï¼the group with TG+GC performed better than the group with GCï¼RR=1.27ï¼95%CI[1.21ï¼1.34]ï¼ï¼and the group with CTX+GC performed better than the group with TG+GCï¼RR=0.60ï¼95%CI[0.43ï¼0.85]ï¼. No significant difference was found between the group with TG+GC and LEF+GCï¼RR=0.68ï¼95%CI[0.30ï¼1.53]ï¼. In terms of urinary protein, urine occult blood negative timeï¼the group with TG performed better than the group with conventional therapyï¼MD=-9.00ï¼95% CI[-11.99ï¼-6.01];MD=-12.00ï¼95%CI[-16.13ï¼-7.87]ï¼ï¼the group with TG+GC performed better than the group with GCï¼MD=-8.86ï¼95%CI[-10.08ï¼-7.64];MD=-16.24ï¼95%CI[-23.80ï¼-8.67]ï¼. In terms of recurrence rate, the group with TG+GC was lower than the group with GCï¼RR=0.13ï¼95%CI[0.06ï¼0.25]ï¼, but there were no significant difference between the group with TG and conventional therapyï¼RR=0.43ï¼95%CI[0.15ï¼1.19]ï¼. In adverse reactions, the common adverse effects of TG were gastrointestinal discomfort, liver damage and leucopenia. TG for the treatment of HSPN can improve clinical efficacy, reduce recurrence, and the adverse reactions are relatively safe. Due to the generally low methodological quality of the included studies, which affected the accuracy and reliability of the result. Therefore, more high-quality, large samples and multi-center randomized controlled trials are necessary for further evidence.
Assuntos
Vasculite por IgA , Tripterygium , Glicosídeos , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto , Reprodutibilidade dos TestesRESUMO
As a structural analogue of pyridylthiazole, 2-(2-benzothiazoyl)-phenylethynylquinoline (QBT) was designed as a fluorescent probe for Hg(II) based on an intramolecular charge transfer (ICT) mechanism. The compound was synthesized in three steps starting from 6-bromo-2-methylquinoline, with moderate yield. Corresponding studies on the optical properties of QBT indicate that changes in the fluorescence ratio of QBT in response to Hg(II) could be quantified based on dual-emission changes. More specifically, the emission spectrum of QBT before and after interactions with Hg(II) exhibited a remarkable red shift of about 120 nm, which is rarely reported in ICT-based fluorescent sensors. Finally, QBT was applied in the two-channel imaging of Hg(II) in live HeLa cells.
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Corantes Fluorescentes/química , Mercúrio/análise , Imagem Óptica , Sobrevivência Celular , Corantes Fluorescentes/síntese química , Células HeLa , Humanos , Estrutura Molecular , Teoria Quântica , Espectrometria de FluorescênciaRESUMO
Follicle-stimulating hormone (FSH) has key roles in animal reproduction, including spermatogenesis and ovarian maturation. Many factors influence FSH secretion. However, despite the broad functions of microRNAs (miRNAs) via the regulation of target genes, little is known about their roles in FSH secretion. Our previous results suggested that miR-186-5p targets the 3' UTR of FSHb; therefore, we examined whether miR-186-5p could regulate FSH secretion in rat anterior adenohypophyseal cells. miR-186-5p was transfected into rat anterior pituitary cells. The expression of FSHb and the secretion of FSH were examined by RT-qPCR and ELISA. A miR-186-5p mimic decreased the expression of FSHb compared with expression in the control group and decreased FSH secretion. In contrast, both the mRNA levels and secretion of FSH increased in response to miR-186-5p inhibitors. Our results demonstrate that miR-186-5p regulates FSH secretion by directly targeting the FSHb 3' UTR, providing additional functional evidence for the importance of miRNAs in the regulation of animal reproduction.
Assuntos
Subunidade beta do Hormônio Folículoestimulante/genética , Hormônio Foliculoestimulante/metabolismo , Gonadotrofos/metabolismo , MicroRNAs/metabolismo , Reprodução/genética , Regiões 3' não Traduzidas/genética , Animais , Células Cultivadas , Masculino , MicroRNAs/antagonistas & inibidores , Mutagênese , Cultura Primária de Células , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , TransfecçãoRESUMO
Many long non-coding RNAs (lncRNAs) have been identified in several types of human pituitary adenomas and normal anterior pituitary, some of which are involved in the pathogenesis of pituitary adenomas. However, a systematic analysis of lncRNAs expressed at different developmental stages of normal pituitary, particularly in rats, has not been performed. Therefore, we contrasted two cDNA libraries of immature (D15) and mature (D120) anterior pituitary in rat that were sequenced on an Illumina HiSeq Xten platform, and a total of 29,568,806,352 clean reads were identified. Notably, 7039 lncRNA transcripts corresponded to 4442 lncRNA genes, and 1181 lncRNA transcripts were significantly differentially expressed in D15 and D120. In addition, 6839 protein-coding genes (<100 kb upstream and downstream) were the nearest neighbors of 4074 lncRNA genes. An interaction network of lncRNAs and the follicle-stimulating hormone beta-subunit (FSHb) gene was constructed using the lncRNATargets platform, and three novel lncRNAs were obtained. Furthermore, we detected the expression of the novel lncRNAs and ten highly expressed lncRNAs that were randomly selected through quantitative PCR (qPCR). The rat anterior pituitary lncRNA content identified in this study provides a more in-depth understanding of the roles of these lncRNAs in hormone and reproduction development and regulation in mammals.
Assuntos
Hipófise/metabolismo , Neoplasias Hipofisárias/genética , RNA Longo não Codificante/genética , Animais , Perfilação da Expressão Gênica/métodos , Biblioteca Gênica , Redes Reguladoras de Genes/genética , Ratos , Ratos WistarRESUMO
Emerging evidence suggests that dipeptidyl peptidase-4 (DPP-4) inhibitors, including sitagliptin, exert favourable effects on the vascular endothelium. DPP-4 inhibitors suppress the degradation of glucagon-like peptide-1 (GLP1), which has been reported to enhance nitric oxide (NO) production. However, the effects of DPP-4 inhibitors on endothelin-1 (ET-1) expression in the aorta, as well as the underlying mechanisms responsible for these effects, have yet to be investigated in animal models of diabetes mellitus (DM). In the present study, the rats were randomly divided into the following four groups: i) control; ii) DM; iii) DM + lowdose sitagliptin (10 mg/kg); and iv) DM + highdose sitagliptin (30 mg/kg). Apart from the control group, all the rats received a high-fat diet for 8 weeks prior to the induction of diabetes with an intraperitoneal injection of streptozotocin. The treatments were then administered for 12 weeks. The serum levels of ET-1, NO, GLP-1 and insulin were measured as well as endothelial function. The expression of ET-1, AMP-activated protein kinase (AMPK) and nuclear factor (NF)-κB/IκBα were determined. After 12 weeks of treatment, the diabetic rats receiving sitagliptin showed significantly elevated serum levels of GLP-1 and NO, and reduced levels of ET-1. Moreover, sitagliptin significantly attenuated endothelial dysfunction as well as the remodeling of the aortic wall. Notably, sitagliptin inhibited ET-1 expression at the transcriptional and translational level in the aorta, which may have been mediated by the suppression of the NF-κB/IκBα system induced by AMPK activation. The majority of the above-mentioned effects were dose dependent. Taken together, the findings of the present study indicate that sitagliptin inhibits ET-1 expression in the aortic endothelium by suppressing the NF-κB/IκBα system through the activation of the AMPK pathway in diabetic rats. These findings further demonstrate some of the vasoprotective properties of DPP-4 inhibitors in vivo.
Assuntos
Arteriosclerose/tratamento farmacológico , Diabetes Mellitus Experimental/tratamento farmacológico , Inibidores da Dipeptidil Peptidase IV/farmacologia , Hipoglicemiantes/farmacologia , Quinase I-kappa B/antagonistas & inibidores , NF-kappa B/antagonistas & inibidores , Fosfato de Sitagliptina/farmacologia , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Aorta/patologia , Arteriosclerose/etiologia , Arteriosclerose/genética , Arteriosclerose/patologia , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Dieta Hiperlipídica/efeitos adversos , Relação Dose-Resposta a Droga , Endotelina-1/genética , Endotelina-1/metabolismo , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Regulação da Expressão Gênica , Peptídeo 1 Semelhante ao Glucagon/antagonistas & inibidores , Peptídeo 1 Semelhante ao Glucagon/genética , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Masculino , NF-kappa B/genética , NF-kappa B/metabolismo , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/biossíntese , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , EstreptozocinaRESUMO
Interaction between advanced glycation endproducts (AGEs) and receptor for AGEs (RAGE) as well as downstream pathways leads to vascular endothelial dysfunction in diabetes. Glucagon-like peptide-1 (GLP-1) has been reported to attenuate endothelial dysfunction in the models of atherosclerosis. However, whether GLP-1 exerts protective effects on aortic endothelium in diabetic animal model and the underlying mechanisms are still not well defined. Experimental diabetes was induced through administration with combination of high-fat diet and intraperitoneal injection of streptozotocin. Rats were randomly divided into four groups, including controls, diabetes, diabetes + sitagliptin (30 mg/kg/day), diabetes + exenatide (3 µg/kg/12 h). Eventually, endothelial damage, markers of inflammation and oxidative stress, were measured. After 12 weeks administration, diabetic rats received sitagliptin and exenatide showed significant elevation of serum NO level and reduction of ET-1 as well as inflammatory cytokines levels. Moreover, sitagliptin and exenatide significantly inhibited aortic oxidative stress level and improved aortic endothelial function in diabetic rats. Importantly, these drugs inhibited the protein expression level in AGE/RAGE-induced RhoA/ROCK/NF-κB/IκBα signaling pathways and activated AMPK in diabetic aorta. Finally, the target proteins of p-eNOS, iNOS, and ET-1, which reflect endothelial function, were also changed by these drugs. Our present study indicates that sitagliptin and exenatide administrations can improve endothelial function in diabetic aorta. Of note, RAGE/RhoA/ROCK and AMPK mediated NF-κB signaling pathways may be the intervention targets of these drugs to protect aortic endothelium.