Project description:The PI3K-AKT pathway is known to regulate cytokines in dust mite-induced pediatric asthma. However, the underlying molecular steps involved are not clear. In order to clarify further the molecular steps, this study investigated the expression of certain genes and the involvement of miRNAs in the PI3K-AKT pathway, which might affect the resultant cytokine-secretion. In-vivo and in-vitro ELISA, qRT-PCR, western-blot and microarrays analyses were used in this study. A down-expression of miRNA-27b-3p in dust mite induced asthma group (group D) was found by microarray analysis. This was confirmed by qRT-PCR that found the miRNA-27b-3p transcripts that regulated the expression of SYK and EGFR were also significantly decreased (p < 0.01) in group D. The transcript levels of the SYK and PI3K genes were higher, while those of EGFR were lower in the former group. Meanwhile, we found significant differences in plasma concentrations of some cytokines between the dust mite-induced asthma subjects and the healthy controls. On the other hand, this correlated with the finding that the transcripts of SYK and its downstream PI3K were decreased in HBE transfected with miRNA-27b-3p, but were increased in HBE transfected with the inhibitor in vitro. Our results indicate that the differential expression of the miRNAs in dust mite-induced pediatric asthma may regulate their target gene SYK and may have an impact on the PI3K-AKT pathway associated with the production of cytokines. These findings should add new insight into the pathogenesis of pediatric asthma.
Project description:The PI3K-AKT pathway is known to regulate cytokines in dust mite-induced pediatric asthma. However, the underlying molecular steps involved are not clear. In order to clarify further the molecular steps, this study investigated the expression of certain genes and the involvement of miRNAs in the PI3K-AKT pathway, which might affect the resultant cytokine-secretion. in-vivo and in-vitro ELISA, qRT-PCR and microarrays analyses were used in this study. A down-expression of miRNA-27b-3p in dust mite induced asthma group (group D) was found by microarray analysis. This was confirmed by qRT-PCR that found the miRNA-27b-3p transcripts that regulated the expression of SYK and EGFR were also significantly decreased (p < 0.01) in group D. The transcript levels of the SYK and PI3K genes were higher, while those of EGFR were lower in the former group. Meanwhile, we found significant differences in plasma concentrations of some cytokines between the dust mite-induced asthma subjects and the healthy controls. On the other hand, this correlated with the finding that the transcripts of SYK and its downstream PI3K were decreased in HBE transfected with miRNA-27b-3p, but were increased in HBE transfected with the inhibitor in vitro. Our results indicate that the differential expression of the miRNAs in dust mite-induced pediatric asthma may regulate their target gene SYK and may have an impact on the PI3K-AKT pathway associated with the production of cytokines. These findings should add new insight into the pathogenesis of pediatric asthma.
Project description:CD4 T cells are essential mediators of the asthmatic process. We used the clinically relevant allergen house dust mites to induce signs of allergy in mice and performed gene expression arrays specifically on CD4 T cells infiltrating the lung Reference: IL-21-producing CD4+ T cells promote type 2 immunity to house dust mites Primary CD4+ T cells were isolated from mice sensitised and challenged to either house dust mites or PBS. Purification of CD4 T cells was performed by flow cytometry. RNA was isolated, converted to cDNA and then hybridised on Affymetrix GeneChip Mouse Gene 2.0 ST Arrays
Project description:CD4 T cells are essential mediators of the asthmatic process. We used the clinically relevant allergen house dust mites to induce signs of allergy in mice and performed gene expression arrays specifically on CD4 T cells infiltrating the lung Reference: IL-21-producing CD4+ T cells promote type 2 immunity to house dust mites
Project description:Aim: The heart undergoes pathological remodelling under increased stress and neuronal imbalance. MicroRNAs (miRNAs) are involved in post-transcriptional regulation of genes in cardiac physiology and pathology. However, the mechanisms underlying miRNA-mediated regulation of pathological cardiac remodelling remain to be studied. This study aims to explore the function of endogenous microRNA-27b-3p (miR-27b-3p) in pathological cardiac remodelling. Methods and results: We found that miR-27b-3p expression was elevated in heart of patients with cardiac hypertrophy and in transverse aortic constriction (TAC)-induced cardiac hypertrophy mouse model. MiR-27b-3p-knockout mice showed significantly attenuated cardiac hypertrophy, fibrosis, and inflammation induced by two independent pathological cardiac hypertrophy models, TAC and Angiotensin II (Ang II) perfusion. Transcriptome sequencing analysis revealed that miR-27b-3p deletion significantly downregulated TAC-induced cardiac hypertrophy, fibrosis, and inflammatory genes. We identified fibroblast growth factor 1 (FGF1) as a novel miR-27b-3p target gene in the heart, which was upregulated in miR-27b-3p-null mice. Conclusions: Our study has demonstrated that miR-27b-3p induces pathological cardiac remodelling and suggests that inhibition of endogenous miR-27b-3p or administration of FGF1 might have the potential to suppress cardiac remodelling in a clinical setting.
Project description:Allergic asthma, a chronic disease-causing inflammation in the airways, is a significant public health concern. Our research team discovered that more than 80% of allergic children in central Taiwan were sensitized to various house dust mites (HDMs). This investigation focuses on how the crude extracts of HDMs affect human epithelium BEAS-2B cells to understand the early fundamental mechanisms for preventing allergic diseases. Therefore, RNA-seq analysis revealed that three Dermatophagoides HDMs allergens activate a common Toll-like receptor signaling pathway in human epithelial cells within a 4-hour treatment. During this process, the nuclear transcription factor NF-κB translocated into the cell nucleus within 30 minutes of allergen stimulation, triggering the expression of pro-inflammatory genes such as IL-6 and IL-8 over 4 hours. Additionally, treating the cells with specific Dermatophagoides microceras (Der m) allergens, there was an upregulation of genes in regulating type 1 diabetes mellitus signaling pathways, mediating IL-12A inflammation. Moreover, an increase in gene sets related to cilia function and the microtubule cytoskeleton in human epithelial cells following treatment combined with Der m allergens and Dexamethasone. Furthermore, OMICs analysis investigates the effects of HDMs allergenic stimulation on the human epidermal cells. This process aims to enhance our understanding of the cellular molecular mechanisms underlying potential targets and bioactive substances in precision medicine for treating asthma caused by HDMs allergens.
Project description:OBJECTIVE: To identify the changes in gene expression elicited by miR-27b-3p mimic transfection of human osteoarthritis (OA) primary fibroblast-like synoviocytes (FLS) obtained from patients undergoing knee arthroplasty. Cells were cultured, and treated for 48 h with miR-27b-3p mimic or Cel-miR-39-3p control mimic. Total RNA was then extracted, and mRNA libraries were prepared using Illumina's TruSEQ stranded total RNA library preparation kit, pooled together, and sequenced on Illumina's NextSeq550.
Project description:To identify the microRNA-27b (miR-27b) target genes in luminal-type breast cancer cells, we performed the microarray analysis using miR-27b knockdown MCF7-luc cell line (MCF7-luc anti-miR-27b), miR-27b overexpressing MCF7-luc cell line (MCF7-luc miR-27b o.e.) and their contro cell line (MCF7-luc anti-NC).