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Hypoxic-ischemic brain damage (HIBD) is a familiar neurological disorder. Emerging reports manifest that microRNAs (miRs) are related to the progression of HIBD. The goal of this study is to explore the mechanism of miR-192-5p in HIBD via regulation of Yes-associated protein 1 (YAP1)-mediated Hippo signaling pathway. The miR-192-5p, YAP1, and Hippo pathway-related factors Phospho (p)-Triaminoguanidinium azide (TAZ) in hippocampal tissues and neurons were detected. The regulatory relationship between miR-192-5p and YAP1 was verified. Neonatal hypoxic ischemia and oxygen-glucose deprivation (OGD) were used to simulate HIBD in vivo and in vitro. The neurobehavioral impairment, neuronal damage and vascular endothelial growth factor (VEGF) expression of neonatal rats in each group were detected. The viability, apoptosis and VEGF expression of hippocampal neurons in each group were also examined. MiR-192-5p expression was elevated while YAP1 expression was reduced in hippocampal tissues of HIBD rats in vivo and OGD neurons in vitro. MiR-192-5p had a targeting relation with YAP1. Suppressed miR-192-5p or overexpressed YAP1 in HIBD rats alleviated neurobehavioral impairment and neuronal damage, and decreased the expression levels of p-TAZ and VEGF expression in vivo. Reduced miR-192-5p or augmented YAP1 decelerated the neuron apoptosis, decreased the p-TAZ level and VEGF level and promoted cell viability of OGD hippocampal neurons in vitro. The study highlights that inhibited miR-192-5p protects against HIBD via regulation of YAP1 and Hippo signaling pathway, which is beneficial for HIBD treatment.
Assuntos
Hipóxia-Isquemia Encefálica , MicroRNAs , Animais , Animais Recém-Nascidos , Apoptose/fisiologia , Modelos Animais de Doenças , Via de Sinalização Hippo , Hipocampo/metabolismo , Hipóxia-Isquemia Encefálica/metabolismo , MicroRNAs/metabolismo , Ratos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas de Sinalização YAPRESUMO
BACKGROUND: The incidence of liver fibrosis remains high due to the lack of effective therapies. Our previous work found that microRNA (miR)-34a expression was increased, while acy1-CoA synthetase long-chain family member1 (ACSL1) was decreased, in a dimethylnitrosamine (DNS)-induced hepatic fibrosis rat model. We hypothesized that miR-34a may play a role in the process of hepatic fibrosis by targeting ACSL1. MATERIAL AND METHODS: From days 2 to 14, cultured primary hepatic stellate cells (HSCs) underwent cell morphology, immunocytochemical staining, and quantitative reverse transcription PCR (RT-qPCR) for alpha smooth muscle actin (a-SMA), desmin, rno-miR-34a, and ACSL1 expression. Wild-type and mutant luciferase reporter plasmids were constructed according to the predicted miR-34a binding site on the 3'-untranslated region (UTR) of the ACSL1 mRNA and then transfected into HEK293 cells. rno-miR-34a was silenced in HSCs to confirm that rno-miR-34a negatively regulates ACSL1 expression. mRNA and protein expression of α-SMA, type I collagen, and desmin were assayed in miR-34a-silenced HSCs. RESULTS: HSCs were deemed quiescent during the first 3 days and activated after 10 days. rno-miR-34a expression increased, and ACSL1 expression decreased, from day 2 to 7 to 14. rno-miR-34a was shown to specifically bind to the 3'-UTR of ACSL1. miR-34a-silenced HSCs showed higher ACSL1and lower α-SMA, type I collagen, and desmin expression than that of matching negative controls and non-transfected cells. CONCLUSIONS: miR-34a appears to play an important role in the process of liver fibrosis by targeting ACSL1 and may show promise as a therapeutic molecular target for hepatic fibrosis.
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Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Regulação da Expressão Gênica , Células Estreladas do Fígado/citologia , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Actinas/metabolismo , Animais , Dimetilnitrosamina , Inativação Gênica , Células HEK293 , Humanos , Imuno-Histoquímica , Fígado/efeitos dos fármacos , Fígado/patologia , Cirrose Hepática/patologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Knowledge on natural background levels (NBLs) of aluminum (Al) in groundwater can accurately assess groundwater Al contamination at a regional scale. However, it has received little attention. This study used a combination of preselection and statistic methods consisting of the oxidation capacity and the boxplot iteration methods to evaluate the NBL of shallow groundwater Al in four groundwater units of the Pearl River Delta (PRD) via eliminating anthropogenic-impacted groundwaters and to discuss driving factors controlling high NBLs of Al in groundwater in this area. A total of 280 water samples were collected, and 18 physico-chemical parameters including Redox potential, dissolved oxygen, pH, total dissolved solids, HCO3 -, NH4 +, NO3 -, SO4 2-, Cl-, NO2 -, F-, K+, Na+, Ca2+, Mg2+, Fe, Mn, and Al were analyzed. Results showed that groundwater Al NBLs in groundwater units A-D were 0.11, 0.16, 0.15, and 0.08 mg/L, respectively. The used method in this study is acceptable for the assessment of groundwater Al NBLs in the PRD, because groundwater Al concentrations in various groundwater units in residual datasets were independent of land-use types, but they were opposite in the original datasets. The dissolution of Al-rich minerals in sediments/rocks was the major source for groundwater Al NBLs in the PRD, and the interaction with Al-rich river water was secondary one. The high groundwater Al NBL in groundwater unit B was mainly attributed to the acid precipitation and the organic matter mineralization inducing the release of Al in Quaternary sediments. By contrast, the high groundwater Al NBL in groundwater unit C mainly was ascribed to the release of Al complexes such as fluoroaluminate from rocks/soils into groundwater induced by acid precipitation, but it was limited by the dissolution of Mg minerals (e.g., dolomite) in aquifers. This study provides not only useful groundwater Al NBLs for the evaluation of groundwater Al contamination but also a reference for understanding the natural geochemical factors controlling groundwater Al in urbanized deltas such as the PRD. PRACTITIONER POINTS: The natural background level (NBL) of groundwater aluminum in the Pearl River Delta (PRD) was evaluated. The dissolution of aluminum-rich minerals in sediments/rocks was the major source for groundwater aluminum NBLs in the PRD. The acid precipitation and organic matter mineralization contribute to high groundwater Al NBL in the groundwater unit B. The acid precipitation contributes to high groundwater Al NBL in the groundwater unit C, while dissolution of magnesium minerals limits it.
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Alumínio , Monitoramento Ambiental , Água Subterrânea , Poluentes Químicos da Água , Água Subterrânea/química , Água Subterrânea/análise , Alumínio/análise , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/química , Rios/química , China , UrbanizaçãoRESUMO
Alzheimer's disease (AD) is a type of neurodegenerative disease characterized by cognitive impairment that is aggravated with age. The pathological manifestations include extracellular amyloid deposition, intracellular neurofibrillary tangles and loss of neurons. As the world population ages, the incidence of AD continues to increase, not only posing a significant threat to the well-being and health of individuals but also bringing a heavy burden to the social economy. There is epidemiological evidence suggesting a link between AD and metabolic diseases, which share pathological similarities. This potential link would deserve further consideration; however, the pathogenesis and therapeutic efficacy of AD remain to be further explored. The complex pathogenesis and pathological changes of AD pose a great challenge to the choice of experimental animal models. To understand the role of metabolic diseases in the development of AD and the potential use of drugs for metabolic diseases, the present article reviews the research progress of the comorbidity of AD with diabetes, obesity and hypercholesterolemia, and summarizes the different roles of animal models in the study of AD to provide references for researchers.
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Stroke has become the most common cause of death among residents in China, among which ischemic stroke accounts for the vast majority reaching 70% to 80%. It is of great importance to actively investigate the protective mechanism of cerebral ischemia injury after IS (ischemic stroke). We constructed cerebral ischemia injury models in vivo MACO rat and in vitro (oxygen-glucose deprivation cell model) and set up different interference groups. RT-PCR (reverse transcription PCR) was conducted to detect the expression of lncRNA in neuronal cells, brain tissue, and plasma of different groups, and ELISA (enzyme-linked immunosorbent assay) and western blot were used to detect the expression of the protein in neuronal cells, brain tissue, and plasma of different groups. Cell activity was detected by the CCK-8 assay, while cell apoptosis was examined by TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay. In the rats' neuronal cells and brain tissue, curcumin can inhibit the expression of lncRNA GAS5 (long noncoding RNA growth arrest-specific 5). In oxygen-glucose-deprived neuronal cells in vitro, curcumin and low-expressed lncRNA GAS5 can enhance cell activity and decline cell apoptosis, but the addition of curcumin and overexpressed lncRNA GAS5 can make this phenomenon disappear. In neuronal cells, plasma, and brain tissue, curcumin and the low-expressed lncRNA GAS5 can inhibit the expression of IL-1ß (interleukin 1 beta), TNF-α (tumor necrosis factor alpha), IL-6 (interleukin 6), Sox2 (SRY-box transcription factor 2), Nanog, and Oct4 (octamer-binding transcription factor 4). However, overexpressed lncRNA GAS5 and curcumin made the inhibitory effect disappear. In conclusion, this study demonstrated that curcumin could inhibit the expression of lncRNA GAS5, thereby inhibiting the expression of inflammation-related factors IL-1ß, TNF-α, and IL-6, and ultimately achieve the purpose of attenuating cerebral ischemic cell damage. However, curcumin and lncRNA GAS5 may not alleviate cerebral ischemic cell damage by affecting stem cell differentiation.
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The aim of this study was to investigate the effect of vestibular disruption on autophagy-related proteins and the tumour-associated pathway P13K/Akt in rat sleep and its hypothalamus tissue and to examine whether catechins trigger tumour autophagy. Healthy adult male rats were randomly selected and divided into the vestibular damage group, the sham operation group, and the control group, with 8 rats in each group. A vestibular damage model was established through penetrating the tympanic membrane of the external auditory canal by injecting sodium p-aminophenylarsonate. The electroencephalogram (EGG) activity was used to record the sleep-wakefulness cycle of rats, and the expression levels of hypothalamic orexin (orexin) mRNA and autophagy proteins were detected. Primary hippocampal neurons were intervened with orexin at different concentrations and at different times to detect cell viability and the expression of autophagy protein and P13K/Akt signal pathway protein. The results showed that compared with the control group and the sham operation group, NREM duration in the vestibular damage group decreased significantly (P < 0.05), while its W time increased significantly (P < 0.05). The expression level of orexin mRNA in the hypothalamus of the vestibular damage group was significantly higher than that of the other two groups (P < 0.05), the expression of autophagy microtubule-related proteins LC3B and Beclin-1 increased significantly (P < 0.05), and the protein expression level of p62 decreased significantly (P < 0.05). After orexin intervention, compared with the control group, the expression of Beclin-1 protein that positively correlated with autophagy decreased significantly (P < 0.05) and the expression of mTOR, PDK1, and Akt protein increased significantly (P < 0.05). Compared with the orexin intervention group, the expression of Beclin-1 and LC3B proteins in cells of the orexin receptor inhibitor (Almorexant) group, the autophagy activator (Rapamycin) group, the orexin + Almorexant group, and the orexin + Rapamycin group increased significantly (P < 0.05), and the expression of mTOR, PDK1, and Akt proteins decreased significantly (P < 0.05). Catechins trigger autophagy in part by regulating the p-Akt/p-mTOR and P13K pathways and by stimulating the MAPK pathway. Catechins initiate apoptosis in common tumour types of hepatocellular carcinoma cells by activating autophagy-related pathways. The conclusion is that vestibular damage can affect the sleep-wakefulness cycle of rats; the level of autophagy in hypothalamic tissue is upregulated and may affect cell proliferation and activity through mTOR-P13K/Akt, which has a certain reference value for tumor formation and provides a basis for the research of insomnia or sleep disorders caused by tumors. Autophagy activation is a key process by which catechins promote apoptosis in tumour cells, providing an avenue for more research on the use of catechins-rich diets for cardiovascular protection in the treatment of tumours.
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AIM: Cerebral ischemic injury is one of the debilitating diseases showing that inflammation plays an important role in worsening ischemic damage. Therefore, studying the effects of some potential anti-inflammatory compounds can be very important in the treatment of cerebral ischemic injury. METHODS: This study investigated anti-inflammatory effects of triblock copolymer nanomicelles loaded with curcumin (abbreviated as NC) in the brain of rats following transient cerebral ischemia/reperfusion (I/R) injury in stroke. After preparation of NC, their protective effects against bilateral common carotid artery occlusion (BCCAO) were explored by different techniques. Concentrations of free curcumin (C) and NC in liver, kidney, brain, and heart organs, as well as in plasma, were measured using a spectrofluorometer. Western blot analysis was then used to measure NF-κB-p65 protein expression levels. Also, ELISA assay was used to examine the level of cytokines IL-1ß, IL-6, and TNF-α. Lipid peroxidation levels were assessed using MDA assay and H&E staining was used for histopathological examination of the hippocampus tissue sections. RESULTS: The results showed a higher level of NC compared to C in plasma and organs including the brain, heart, and kidneys. Significant upregulation of NF-κB, IL-1ß, IL-6, and TNF-α expressions compared to control was observed in rats after induction of I/R, which leads to an increase in inflammation. However, NC was able to downregulate significantly the level of these inflammatory cytokines compared to C. Also, the level of lipid peroxidation in pre-treated rats with 80mg/kg NC was significantly reduced. CONCLUSION: Our findings in the current study demonstrate a therapeutic effect of NC in an animal model of cerebral ischemia/reperfusion (I/R) injury in stroke through the downregulation of NF-κB-p65 protein and inflammatory cytokines.
Assuntos
Isquemia Encefálica/tratamento farmacológico , Inflamação/tratamento farmacológico , Micelas , NF-kappa B/metabolismo , Nanopartículas/química , Polímeros/química , Transdução de Sinais , Animais , Anti-Inflamatórios/farmacologia , Encéfalo/patologia , Isquemia Encefálica/sangue , Isquemia Encefálica/complicações , Curcumina/farmacologia , Citocinas/sangue , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Inflamação/complicações , Inflamação/patologia , Lactatos/química , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Nanopartículas/ultraestrutura , Fosforilação/efeitos dos fármacos , Polietilenoglicóis/química , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Distribuição Tecidual/efeitos dos fármacos , Fator de Transcrição RelA/metabolismoRESUMO
BACKGROUND AND AIMS: Liver stiffness measurement (LSM) detected by FibroScan, combined with biochemical indexes, has shown potential values for assessment of liver fibrosis pathological degrees. Here we aimed to investigate a noninvasive method for hepatitis B virus-related liver fibrosis. PATIENTS AND METHODS: In all, 307 patients who underwent liver biopsy and LSM measurement were included. Inflammation grades and fibrosis stages were evaluated according to METAVIR scoring system. Spearman's rank correlation analysis, logistic regression analysis, and receiver operating characteristic (ROC) curves analysis were performed to assess the factors' role in inflammation grades/fibrosis stages. RESULTS: Spearman's rank correlation analysis showed that LSM, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and AST-to-platelet ratio index were positively correlated with inflammation grades and histologic fibrosis stages; platelets showed negative correlation, and AST-to-ALT ratio was not related. Logistic regression analysis indicated that LSM and APRI were risk factors for inflammation grades; LSM was the independent risk factor for fibrosis stages, P<0.0001, odds ratio>1. ROC curve analysis found LSM cutoff values and areas under the curve for the diagnosis of fibrosis scores: 6.95 and 0.804, respectively, for the diagnosis of significant fibrosis (F≥F2); 10.35 and 0.856, respectively, for severe fibrosis (F≥F3); 11.35 and 0.897, respectively, for cirrhosis (F=F4). Considering ALT as a confounding factor, ROC analysis was repeated in patients with normal and elevated ALT separately; the results indicated that when ALT was up to 40 U/l, LSM cutoff value and areas under the curve for the diagnosis of significant fibrosis (F≥F2) were 6.55 and 0.748, respectively. CONCLUSION: This study provided a noninvasive treatment and prevention indicator for early hepatitis B virus-related liver fibrosis.
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Técnicas de Imagem por Elasticidade , Hepatite B Crônica/diagnóstico por imagem , Hepatite B Crônica/virologia , Cirrose Hepática/diagnóstico por imagem , Fígado/diagnóstico por imagem , Adulto , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Biomarcadores/sangue , Biópsia , Plaquetas , Ensaios Enzimáticos Clínicos , Diagnóstico Precoce , Feminino , Hepatite B Crônica/sangue , Hepatite B Crônica/patologia , Humanos , Fígado/enzimologia , Fígado/patologia , Fígado/virologia , Cirrose Hepática/sangue , Cirrose Hepática/patologia , Cirrose Hepática/virologia , Masculino , Pessoa de Meia-Idade , Contagem de Plaquetas , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Índice de Gravidade de DoençaRESUMO
Early diagnosis of liver fibrosis is critical for early intervention and prognosis of various chronic liver diseases. Conventional repeated histological assessment is impractical due to the associated invasiveness. In the current study, we evaluated circulating miR-185 as a potential biomarker to predict initiation and progression of liver fibrosis. We found that miR-185 was significantly up-regulated in blood specimens from patients with HBV-liver fibrosis and rats with liver fibrosis, the miR-185 levels were correlated with liver fibrosis progression, but not with the different viral loads in HBV-infected patients. miR-185 was observed in collagen deposition regions during advanced liver fibrosis. We found that differences in miR-185 levels facilitated the discrimination between early-staged or advanced-staged liver fibrosis and the healthy controls with high specificity, sensitivity, and likelihood ratio using receiver-operator characteristic analysis. miR-185 targeted SREBF1, and increased expression of COL1A1 and a-SMA genes that are hallmarks of liver fibrosis. Our data supported that circulating miR-185 levels could be used as potential biomarkers for the early diagnosis of liver fibrosis.