RESUMO
PURPOSE: To study neurochemical reactions to chronic intermittent hypoxia (CIH) in the hypoglossal nucleus (HN) of rats. METHODS: Adult male Sprague-Dawley rats (n = 12) were randomly divided into two groups (the CIH and the control group). The CIH rats were housed in a hypoxic chamber with the fraction of oxygen volume alternating between 21% and 5% by providing air for 60 s and then providing nitrogen for 60 s from 8:30 am to 16:30 pm each day for 35 days. The control group was housed in a cabin with normal oxygen levels. We studied the expression of c-fos protein, 5-hydroxytryptamine (5-HT) positive terminals, and its 2A receptors in hypoglossal nuclei by immunohistochemistry. RESULTS: The expression of c-fos, 5-HT positive terminals, and accordingly 5-HT 2A receptors in the CIH group were significantly higher than that in the controls (p < 0.05). The ventral side of the HN showed a clearly higher expression of 5-HT and its 2A receptors than the dorsal side (p < 0.05). CONCLUSION: There were 2 responses of the HN to CIH. First, CIH induced a higher expression of 5-HT positive terminals and its 2A receptors, and second, this reaction was much more evident in ventral side than in the dorsal side. We postulate that these responses may serve to be a protective and compensatory mechanism for CIH.
Assuntos
Nervo Hipoglosso/metabolismo , Hipóxia/metabolismo , Bulbo/metabolismo , Animais , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Proteínas Proto-Oncogênicas c-fos/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Receptor 5-HT2A de Serotonina/metabolismo , Serotonina/metabolismoRESUMO
The gut microbiota is crucial in the pathogenesis of type 2 diabetes mellitus (T2DM). However, the metabolism of T2DM patients is not well-understood. We aimed to identify the differences on composition and function of gut microbiota between T2DM patients with obesity and healthy people. In this study, 6 T2DM patients with obesity and 6 healthy volunteers were recruited, and metagenomic approach and bioinformatics analysis methods were used to understand the composition of the gut microbiota and the metabolic network. We found a decrease in the abundance of Firmicutes, Oribacterium, and Paenibacillus; this may be attributed to a possible mechanism and biological basis of T2DM; moreover, we identified three critical bacterial taxa, Bacteroides plebeius, Phascolarctobacterium sp. CAG207, and the order Acidaminococcales that can potentially be used for T2DM treatment. We also revealed the composition of the microbiota through functional annotation based on multiple databases and found that carbohydrate metabolism contributed greatly to the pathogenesis of T2DM. This study helps in elucidating the different metabolic roles of microbes in T2DM patients with obesity.
Assuntos
Bactérias/classificação , Diabetes Mellitus Tipo 2/microbiologia , Microbioma Gastrointestinal , Metagenoma , Obesidade/microbiologia , Adulto , Bactérias/metabolismo , Biologia Computacional , Diabetes Mellitus Tipo 2/fisiopatologia , Fezes/microbiologia , Feminino , Voluntários Saudáveis , Humanos , Masculino , Metagenômica , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Previous studies confirmed that the intronic miRNAs participated in regulating host gene-primed biological processes. The coordinated roles of miR-28 with its host gene, LIM domain lipoma-preferred partner (LPP), remain unknown in atherosclerosis. METHODS: In this study, we determined to assess circulating levels of miR-28-5p in unstable angina patients, compared with age- and sex- matched control subjects by quantitative PCR. Furthermore, we attempted to explore whether miR-28-5p could influence the expression of ATP-binding cassette transporter A1 (ABCA1) and liver X receptor (LXR), major mediators of high density lipoprotein (HDL) synthesis and transportation in hepatic cells and macrophages. RESULTS: It was found that plasma levels of miR-28-5p were significantly increased in unstable angina patients with or without type 2 diabetes mellitus. Notably, miR-28-5p upregulated ABCA1 expression at transcription and translation levels, strongly correlated with translational activation of LXRα in HepG2 and THP-1-derived macrophages. CONCLUSIONS: Our findings suggest that circulating miR-28-5p, involved in LXRα-ABCA1 pathway, may be a potential biomarker for diagnosis and prognosis of unstable angina.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/sangue , Angina Instável/sangue , MicroRNAs/sangue , Receptores Nucleares Órfãos/sangue , Transdução de Sinais , Idoso , Angina Instável/diagnóstico , Biomarcadores/sangue , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Feminino , Células Hep G2 , Humanos , Receptores X do Fígado , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
In order to study the molecular differences between type 2 diabetes mellitus (T2DM) and T2DM with depression (DD), we aimed to screen the differential expression of lncRNA, mRNA, and circRNA in the blood of patients with T2DM and DD. Based on the self-rating depression scale (SDS), patient health questionnaire 9 (PHQ9), blood glucose and HbA1c, we divided the patients into T2DM and DD group. Peripheral blood was collected from the two groups of patients to perform lncRNA, mRNA, and circRNA expression profiling and screening DD-related specific molecules. Subsequently, bioinformatics analysis was performed to investigate the functions of differentially expressed genes (DEgenes). Finally, RT-PCR and lncRNA-mRNA regulatory network was performed to verify the expressions of lncRNAs and mRNAs related to the occurrence and development of DD. 28 lncRNAs, 107 circRNAs, and 89 mRNAs were identified in DD differential expression profiles. GO and pathway analysis found that 20 biological process (BP) related entities and 20 pathways associated with DD. The analysis shows that the genes that are differentially expressed in the DD group involved in the development of the neuropsychiatric system, immunity, and inflammation. Then, we screening for the important DElncRNA and mRNA associated with DD were verified by RT-PCR experiments and the results of RT-PCR were consistent with the sequencing results. LncRNA, circRNA, and mRNA differential expression profiles exist in DD patients compared with T2DM. The lncRNA-mRNA regulatory network analysis confirmed the crosslinking and complex regulation patterns of lncRNA and mRNA expression and verified the authenticity of the regulatory network.
Assuntos
Depressão/genética , Diabetes Mellitus Tipo 2/genética , Redes Reguladoras de Genes , RNA não Traduzido/genética , Idoso , Depressão/complicações , Diabetes Mellitus Tipo 2/complicações , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA não Traduzido/metabolismo , TranscriptomaRESUMO
By using substractive hybridization (SSH) and rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR), a full-length cDNA encoding Brassica napus small nuclear ribonucleoprotein, named BnSmD1, was obtained. It had 484 base pairs in length containing an open reading frame (ORF) of 354 bp and encoding a predicted protein of 118 amino acids with a molecular weight of 13 kDa. The BnSmD1 protein shares two highly conserved Sm folds (Sm-1 and Sm-2) and a C-terminal RG dipeptide repeat. Northern blot analysis revealed that BnSmD1 was expressed in all tested organs in B. napus, but its transcript level in early floral buds was much higher than that in leaf and stem tissues. No obvious expression difference was observed in leaf and stem tissues between the apetalous line Apet33-10 petalled near-isogenic line Pet33-10. Compared with wild type, the expression of BnSmD1 in the early floral buds of apetalous mutant Apet33-10 was significantly reduced. Taken together, our results suggest that BnSmD1 may play an important role in early floral petal development in B. napus.
Assuntos
Brassica napus/genética , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas Centrais de snRNP/genética , Sequência de Aminoácidos , Western Blotting , Brassica napus/crescimento & desenvolvimento , Clonagem Molecular , Flores/genética , Flores/crescimento & desenvolvimento , Humanos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Proteínas de Plantas/química , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Proteínas Centrais de snRNP/químicaRESUMO
Increasing evidence has suggested that microRNA (miRNA) may play a role in the pathogenesis of cardiovascular disease, which has led to a greater understanding of the complex pathophysiological processes underlying unstable angina (UA). The present study aimed to investigate changes in the miRNA expression profiles of patients with UA using gene-chip analysis, in order to further elucidate the pathogenesis of UA. Total RNA was extracted and purified from plasma samples collected from patients with UA and healthy controls. The samples underwent microarray analysis using an Exiqon miRCURY LNA™ microRNA Array. Differentially expressed miRNAs were identified by volcano plot filtering, and were validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In addition, functional annotation of the differentially expressed miRNAs involved gene ontology analyses. Among the 212 miRNAs differentially expressed between the two groups, 82 were upregulated and 130 were downregulated. Notably, the results of the RT-qPCR were consistent with the gene-chip results. The miRNAs identified in the present study may be potential novel biomarkers for the prevention and early diagnosis of UA. Furthermore, the results of the present study suggested that UA occurs as a result of complex and dynamic processes regulated by numerous factors, including multiple miRNAs.
RESUMO
The biological function of the intronic microRNA-28 (miR-28) may be associated with the biological roles of its host gene, LIM domain lipomapreferred partner (LPP). LPP has been reported to promote smooth muscle cell migration in arterial injury and atherosclerosis. However, the mechanism of miR28 in atherosclerosis remains unclear. In the current study, the aim was to validate the inhibitory effect of miR285p on extracellular signalregulated kinase 2 (ERK2), to investigate its biological role in atherosclerosis and its association with cardiovascular disease. Western blotting and stemloop reverse transcriptionquantitative polymerase chain reaction combined with TaqMAN microRNA analysis was conducted. The current study demonstrated that miR285p upregulated the expression of ATPbinding cassette transporter A1 (ABCA1) via the inhibition of ERK2 in HepG2 cells. In addition, increased levels of plasma miR285p were positively correlated with the levels of highdensity lipoprotein cholesterol in patients with unstable angina. This suggests that miR-28-5p participates in atherosclerosis via ERK2-mediated upregulation of the ABCA1 pathway.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/metabolismo , MicroRNAs/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Angina Instável/sangue , Angina Instável/enzimologia , Angina Instável/genética , Sítios de Ligação , Estudos de Casos e Controles , HDL-Colesterol/sangue , Feminino , Flavonoides/farmacologia , Células Hep G2 , Humanos , Masculino , MicroRNAs/sangue , MicroRNAs/genética , Pessoa de Meia-Idade , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The miR-146-mediated IL-1 receptor-associated kinase 1 (IRAK-1) feedback circuit has been shown to inhibit inflammatory response in macrophages against lipopolysaccharide (LPS). The aim of this study is to compare the antagonized effects of miR-146a-5p on Porphyromonas gingivalis (P.g) lipolysaccharide (LPS)- and advanced glycation end products (AGEs)-triggered ABCA1 and ABCG1 dysregulation and explore the underlying mechanism. THP-1-derived macrophages transfected with miRNA mimics or not were treated with P.g LPS or AGE-BSA, respectively. The mechanism of endotoxin tolerance was mimicked. miR-146a-5p levels and protein levels of IRAK-1, LXRα/ß, ABCA1, and ABCG1 were detected by stem-loop reverse transcription followed by TaqMan PCR analysis and Western blotting. Our results showed that miR-146a-5p levels were significantly increased in macrophages after 24 h of stimulation with high dose of P.g LPS or AGE-BSA. Macrophages transfected with miR-146a-5p mimics attenuated the dysregulation of ABCA1/G1 induced by P.g LPS and AGEs through IRAK-1 downregulation. In low-dose LPS-tolerized cells, elevated miR-146a-5p antagonized the increase of ABCA1, ABCG1, and IRAK-1. However, low-dose AGE-BSA did not increase miR-146a-5p levels. In conclusion, the model of endotoxin tolerance is suitable for the antagonistic effects on the dysregulation of ABCA1/G1 induced by high dose of P.g LPS. Conversely, low-dose AGEs did not induce the model of P.g LPS-mediated tolerance.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/biossíntese , Transportadores de Cassetes de Ligação de ATP/biossíntese , Produtos Finais de Glicação Avançada/toxicidade , Quinases Associadas a Receptores de Interleucina-1/biossíntese , MicroRNAs/biossíntese , Porphyromonas gingivalis , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Animais , Bovinos , Linhagem Celular , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/fisiologia , Humanos , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , MicroRNAs/antagonistas & inibidores , Soroalbumina Bovina/toxicidadeRESUMO
OBJECTIVE: To study the change of TIZ expression in epithelial ovarian cancer cells. METHODS: HO8910 cells were transinfected with siRNA to inhibit the expression of TIZ. pcDNA3.1-TIZ vectors were combined to increase the TIZ expression level. The cell viability, colony forming efficiency and cycle distribution of HO8910, HO8910/NC, HO8910/pcDNA3.1-NC, HO8910/TIZ-573 and H08910/pcDNA3.1-TIZ were compared, and the invasion rate, migration rate and adhesion rate between 5 groups of cells were compared. RESULTS: Compared with those of HO8910, HO8910/NC and HO8910/pcDNA3.1-NC, the cell viability, colony forming efficiency and cell cycle distribution of HO8910/TIZ-573 were increased, while the indexes of H08910/pcDNA3.1-NC were decreased with statistical significant difference (P<0.05). There was no statistical significant difference in the invasion rate, migration rate and adhesion rate between 5 groups of cells (P>0.05). CONCLUSIONS: The expression of TIZ can inhibit the proliferation of epithelial ovarian cancer cells.
RESUMO
Type 2 diabetes mellitus (T2DM) is a complex metabolic disorder caused by the interaction of environmental factors and multiple genes. The genetic background of T2DM is complex and remains to be fully elucidated. MicroRNAs (miRNAs) are negative regulators of gene expression and several miRNAs are associated with the development of T2DM. However, the expression and biological function of miRNA93p in lipid metabolism of patients with T2DM remain to be fully elucidated. The predominant aim of the present study was to examine the effect of miRNA93p on lipid accumulation in HepG2 cells. To investigate this, an MTT assay was used to determine cell proliferation, and the effects of miRNA93p on triglycerides (TG) and total cholesterol (TC) in the HepG2 cells were also examined. Reverse transcriptionquantitative polymerase chain reaction and western blot analyses were used to measure the expression levels of SIRT1 at the gene and protein levels, respectively. The date revealed that downregulation of miRNA93p inhibited the proliferation of HepG2 cells, and significantly reduced the accumulation of lipids, and decreased TG and TC content. In addition, the present study demonstrated that inhibition of miRNA93p increased the protein expression of sirtuin type 1 (SIRT1), but had no effects on the gene expression of SIRT1. Therefore, these findings demonstrated that the inhibition of miRNA93p reduced the proliferation of HepG2 cells and lipid accumulation by upregulating the expression of SIRT1, indicating its potential as a therapeutic target.