RESUMO
OBJECTIVES: Present study aimed to illustrate the role of miR-144-3p in rheumatoid arthritis (RA). METHODS: N1511 chondrocytes were stimulated by interleukin (IL)-1ß to mimic RA injury model in vitro. Rats were subjected to injection of type II collagen to establish an in vivo RA model, and the arthritis index score was calculated. Cell viability was determined by Cell Counting Kit-8. The expression of cartilage extracellular matrix proteins (collagen II and aggrecan) and matrix metalloproteinase protein were determined by quantitative real-time polymerase chain reaction and western blots. Cell apoptosis was measured by flow cytometry. Enzyme-linked immunosorbent assay was applied to test the secretion of pro-inflammatory cytokines (IL-1ß and tumour necrosis factor-α). Tissue injury and apoptosis were detected by haematoxylin-eosin staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling assay staining. Interaction of miR-144-3p and bone morphogenetic protein 2 (BMP2) was verified by dual-luciferase assay. RESULTS: miR-144-3p was dramatically increased in IL-1ß-induced N1511 cells. miR-144-3p depletion elevated cell viability, suppressed apoptosis, pro-inflammatory cytokine releasing, and extracellular matrix loss in IL-1ß-induced N1511 cells. Moreover, miR-144-3p targeted BMP2 to modulate its expression negatively. Activation of phosphatidylinositol 3-kinase (PI3K)/Akt signalling compromised inhibition of BMP2 induced aggravated N1511 cell injury with IL-1ß stimulation. Inhibition of miR-144-3p alleviated cartilage injury and inflammatory in RA rats. CONCLUSION: Collectively, miR-144-3p could aggravate chondrocyte injury inflammatory response in RA via BMP2/PI3K/Akt axis.
Assuntos
Artrite Reumatoide , MicroRNAs , Agrecanas/metabolismo , Animais , Apoptose , Artrite Reumatoide/patologia , Biotina/metabolismo , Proteína Morfogenética Óssea 2/metabolismo , Cartilagem/metabolismo , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , DNA Nucleotidilexotransferase/metabolismo , Amarelo de Eosina-(YS)/metabolismo , Interleucina-1beta/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
PURPOSE: Idiopathic pulmonary fibrosis (IPF) is a type of progressive lung fibrosis disease. The survival time of diagnosed IPF patients is often only 2 years. Currently much evidence showed that the epithelial-mesenchymal transition (EMT) process is the main cause of the occurrence and development of IPF. LncRNA cardiac hypertrophy related factor (CHRF) was reported to be related with IPF development. Here we explored the functions and regulatory mechanisms of CHRF on EMT in IPF. MATERIALS AND METHODS: A549 cells were treated with transforming growth factor-ß1 (TGF-ß1) for 48 h to construct IPF cell model. CHRF and miR-146a expression were quantified using qPCR. The expression of L1 cell adhesion molecule (L1CAM) and EMT related indicators (E-cadherin, Vimentin, Slug and N-cadherin) were detected by qPCR and western blot. Dual luciferase reporter experiment was conducted to prove the molecular interaction of miR-146a and L1CAM, as well as CHRF and miR-146a. RESULTS: CHRF and L1CAM expression were significantly upregulated and promoted the EMT process in A549 after treatment of TGF-ß1. MiR-146a was obviously down-regulated, and knockdown of CHRF inhibited the EMT process by up-regulating miR-146a, in A549 after treatment of TGF-ß1. Meanwhile, overexpression of miR-146a inhibited EMT process via targeting L1CAM. In addition, L1CAM overexpression eliminated the inhibitory effect of sh-CHRF on the EMT process. CONCLUSIONS: These results provided evidence that CHRF promoted EMT process in A549 after treatment of TGF-ß1, which proposed a new insight for depth understanding the pathological mechanisms of IPF.
Assuntos
Transição Epitelial-Mesenquimal , MicroRNAs , Molécula L1 de Adesão de Célula Nervosa , RNA Longo não Codificante , Células Epiteliais Alveolares , Linhagem Celular , Células Epiteliais , Humanos , MicroRNAs/genética , RNA Longo não Codificante/genética , Fator de Crescimento Transformador beta1RESUMO
The domoic acid (DA) produced by certain species of the marine pennate diatom genus Pseudo-nitzschia is highly neurotoxic and can induce nerve excitability and neurotoxicity by binding with ionotropic glutamate receptors, causing amnesic shellfish poisoning in humans who consume seafood contaminated with DA. In recent years, poisoning of humans caused by DA has occurred around the world, which has attracted increasing attention, and studies on DA production by Pseudo-nitzschia have become the hotpot. This article reviews the progress in the biosynthesis of DA by the typical diatom Pseudo-nitzschia, in which the metabolic pathway of the biosynthesis of DA and its precursors, i.e., geranyl pyrophosphate and L-glutamate, and the various environmental factors affecting DA production including temperature, light intensity, nutrients, trace metals, and alien bacteria are discussed. The detection methods of DA (including bioassays, enzyme linked immunosorbent assays, high performance liquid chromatography, capillary electrophoresis and biosensors), as well as the morphology and toxigenicity of Pseudo-nitzschia are also presented.
Assuntos
Diatomáceas , Síndromes Neurotóxicas , Intoxicação por Frutos do Mar , Humanos , Diatomáceas/metabolismo , Ácido Caínico/análise , Ácido Caínico/metabolismo , Redes e Vias MetabólicasRESUMO
Rheumatoid arthritis (RA) often leads to functional disabilities and deformities. MiRNA plays a vital role in cell pyroptosis. Nevertheless, the function and underlying mechanism of miR-144-3p in pyroptosis during the progression of RA remains unclear. In this study, N1511 cells were stimulated with IL-1ß to construct a RA model. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was performed to assess the cell viability. Cell pyroptosis was detected by flow cytometry. The levels of inflammatory cytokines (TNF-α, IL-6, and IL-18) were assessed by enzyme-linked immunosorbent assay (ELISA). The relationship among specific protein 1 (SP1), microRNA-144-3p (miR-144-3p), and phosphatase and tensin homolog (PTEN) was explored by dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP), respectively. The level of miR-144-3p in N1511 cells was upregulated by IL-1ß. MiR-144-3p knockdown inhibited IL-1ß-induced pyroptosis in N1511 cells, and the expressions of NOD-like receptor family pyrin domain containing 3 (NLRP3), Cleaved caspase-1, Gasdermin D (GSDMD), and Cleaved caspase-3 in IL-1ß-stimulated N1511 cells were increased. The levels of inflammatory cytokines in N1511 cells were increased by IL-1ß, which were restored by miR-144-3p knockdown. MiR-144-3p knockdown abolished IL-1ß-induced inactivation of putative kinase 1 (PINK1)/Parkin RBR E3 ubiquitin-protein (Parkin) signalling. Moreover, transcription factor SP1 could upregulate miR-144-3p expression and miR-144-3p negatively regulated PTEN expression. In summary, MiR-144-3p induced by SP1 could promote IL-1ß-induced chondrocyte pyroptosis via inhibiting PTEN expression and suppressing the activation of PINK1/Parkin signalling, which provided a new strategy against RA.
Assuntos
Condrócitos , MicroRNAs , Piroptose , Animais , Linhagem Celular , Condrócitos/metabolismo , Interleucina-1beta , Camundongos , MicroRNAs/genética , PTEN Fosfo-Hidrolase , Proteínas Quinases/metabolismo , Piroptose/genética , Fator de Transcrição Sp1 , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismoRESUMO
Objectives: To retrospectively analyze the correlation between anti-Müllerian hormone (AMH) and the number of oocytes obtained by controlled ovarian hyperstimulation (COH) in women of different ages and explore the factors affecting in vitro fertilization and embryo transfer (IVF-ET) in clinical pregnancy of infertile women to provide evidence for infertile women to choose assisted reproduction strategies. Methods: Infertile women who received IVF-ET or intracytoplasmic sperm injection and embryo transfer (ICSI-ET) treatment in the reproductive center of XX hospital between October 2018 and September 2019 were included. Patient data on medical records, age, body mass index (BMI), years of infertility, basic follicle-stimulating hormone (FSH), basic luteinizing hormone (LH), basic estradiol (E2), anti-Müllerian hormone level (AMH), antral follicle count (AFC), gonadotropins (Gn) medication days, Gn dosage, endometrial thickness on transplantation day, the number of retrieved oocytes, the number of mature oocytes obtained, the number of embryos transferred, clinical pregnancy status, etc., were collected. Results: A total of 314 patients were enrolled in this study, with an average age of 31.0 ± 4.5 years. The infertility period ranged from 0-21 years. The AMH level showed a downward trend with increasing age. Overall, the AMH level of women of all ages was positively correlated with the number of retrieved oocytes (r = 0.335, p < 0.001). The AMH level of women between 22 and 28 years old was positively correlated with the number of retrieved oocytes (r = 0.164, p < 0.061) but it was not statistically significant. Similarly, the AMH level of women aged 29-35 and 36-43 was positively correlated with the number of retrieved oocytes (r = 0.356, p < 0.001; r = 0.461, p < 0.001). The average age of the pregnant group (30.6 ± 4.4 years) was lower than that of the non-pregnant group (32.2 ± 4.6 years) (p < 0.001). The number of oocytes obtained (9.8 ± 4.5) and the number of embryos transferred (1.9 ± 0.4) in the pregnant group was significantly higher than that in the non-pregnant group (9.2 ± 4.5; 1.7 ± 0.5); the difference was statistically significant. The multivariate logistic regression model showed that age (OR = 0.574 95% CI: 0.350-0.940), AMH (OR = 1.430 95% CI: 1.130-1.820) and the number of oocytes obtained (OR = 1.360 95% CI: 1.030-1.790) were factors affecting clinical pregnancy. Conclusion: We found that the level of AMH in infertile women decreased with age and the number of oocytes obtained in infertile women was positively correlated with AMH. Moreover, the number of oocytes and embryo transferred in the pregnant group was significantly higher than those in the non-pregnant group. Furthermore, age, AMH and the number of oocytes affected the clinical pregnancy.