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Effects of anticholinergic agent on miRNAs profile and transcriptome in murine model of allergic rhinitis

ABSTRACT: Purpose: Ipratropium bromide (IB) is able to ameliorate symptoms of allergic rhinitis (AR) by neuroimmunologic mechanism. We intended to explorer whether IB induced effect through modulating specific alteration of miRNA profiling or not. Methods: Nasal sRNAs (miRNAs) profiles of control mice and OVA-induced AR mice that were pre-treated with IB or normal saline before intranasal OVA challenge for 2 or 4 weeks were generated by high-throughput sequencing using Illumina Hiseq 2000.Raw data for sRNA (miRNA) profiles were processed to obtain corresponding clean data which were mapped to reference sequence(mm10) by TopHat(v2.0.9).The expression levels for these resulting mapped miRNAs were estimated by TPM (transcript per million),afterwards, the differentially expressed miRNAs were identified, and the candidate allergic-related miRNAs were further determined by RT-qPCR. Results: Using an optimized data analysis workflow,our data showed that in nasal mucosa of allergic mice with ipratropium bromide (IB) treatment 87 miRNAs are differentially expressed whereas in the nasal mucosa of non-allergic mice 113 miRNAs are differentially expressed, when compared with allergic mice treated with normal saline.IB treatment significantly up-regulated the levels of mmu-miR-124-3p/5p, -133b-5p, -133a-3p/5p, -384-3p, -181a-5p, -378a-5p, -3071-5p. RT-qPCR data further validated these miRNAs expression. Conclusions: Our study represents the detailed analysis of nasal mucosa miRNAs profiles of OVA-induced allergic mice treated with IB, generated by RNA-seq technology. Moreover, IB treatment orchestrated expression of allergic immune-related miRNAs of nasal mucosa in allergic mice, which may associate with ameliorated nasal allergic symptoms

ORGANISM(S): Mus musculus

PROVIDER: GSE92975 | GEO | 2018/07/31

REPOSITORIES: GEO

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Project description:We tested the hypothesis that a panel of placental mammal-specific miRNAs and their targets play important to establish receptivity to implantation and their dysregulated expression may be a feature in women with early pregnancy loss. Relative expression levels of miR-340-5p, −542-3p, and −671-5p all increased following treatment of Ishikawa cells with progesterone (10 μg/ml) for 24 hrs (p < 0.05). RNA sequencing of these P4-treated cells identified co-ordinate changes to 6,367 transcripts of which 1713 were predicted targets of miR-340-5p, 670 of miR-542-3p, and 618 of miR-671-5p. Quantitative proteomic analysis of Ishikawa cells transfected with mimic or inhibitor (48 hrs: n=3 biological replicates) for each of the P4-regulated miRNAs was carried out to identify targets of these miRNAs. Excluding off target effects, mir-340-5p mimic altered 1,369 proteins while inhibition changed expression of 376 proteins (p < 0.05) of which, 72 were common to both treatments. A total of 280 proteins were identified between predicted (mirDB) and confirmed (in vitro) targets. In total, 171 proteins predicted to be targets by mirDB were altered in vitro by treatment with miR-340-5p mimic or inhibitor and were also altered by treatment of endometrial epithelial cells with P4. In vitro targets of miR-542-3p identified 1,378 proteins altered by mimic while inhibition altered 975 a core of 200 proteins were changed by both. 100 protein targets were predicted and only 46 proteins were P4 regulated. miR-671-mimic altered 1,252 proteins with inhibition changing 492 proteins of which 97 were common to both, 95 were miDB predicted targets and 46 were also P4-regulated. All miRNAs were detected in endometrial biopsies taken from patients during the luteal phase of their cycle, irrespective of prior or future pregnancy outcomes Expression of mir-340-5p showed an overall increase in patients who had previously suffered a miscarriage and had a subsequent miscarriage, as compared to those who had infertility or previous miscarriage and subsequently went on to have a life birth outcome. The regulation of these miRNAs and their protein targets regulate the function of transport and secretion, and adhesion of the endometrial epithelia required for successful implantation in humans. Dysfunction of these miRNAs (and therefore the targets they regulate) may contribute to endometrial-derived recurrent pregnancy loss in women.

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Effects of anticholinergic agent on miRNAs profile and transcriptome in murine model of allergic rhinitis

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