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1.
Chem Biodivers ; 2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31943763

RESUMO

Engelhardia roxburghiana Wall. leaves are widely used to develop herbal teas in southeast of China due to medicinal use for diabetes mellitus and hyperlipidemia. Studies have demonstrated that the total flavonoids of Engelhardia roxburghiana Wall. leaves (TFER) exhibited regulatory effects on blood glucose and lipids. To clarify the active ingredients of TFER and their targets in treating atherosclerosis, the present study integrated chemical analysis, network pharmacology analysis and animal experimental studies. Firstly, High Performance Liquid Chromatography - Mass Spectrometry/ Mass Spectrometry (HPLC-MS/MS) was utilized to identify components of TFER. Then active ingredients were screened by Oral bioavailability (OB) and Drug-likeness (DL) index. Thirdly, network was constructed to predict major targets of active ingredients against atherosclerosis. Finally, to verify parts of predicted signaling, Apoe-/- mice were used to develop atherosclerosis. Atherosclerotic plaques in aorta were evaluated by echocardiography. Then serum lipids, target genes expressions in thoracic aorta were determined by qRT-PCR and ELISA methods. Chemical analysis revealed 10 components in TFER sample, 7 of which acted as active ingredients, including naringenin, kaempferol, quercetin, isoengeletin, engeletin, astilbin and quercitrin. KEGG pathway analysis highly enriched in some inflammatory signalings, including NF-κB signaling, Toll-like receptor signaling, TNF signaling. The animal studies indicated TFER reduced atherosclerotic plaques size in aorta, and significantly decreased the serum lipids, down-regulated NF-κB signaling by decreasing mRNA  level of NF-κB p65 subunit , TNF-α and VCAM-1 ,as well as IL-1ß expressions in thoracic aorta, eventually alleviating atherosclerosis progression, which was in consistent with our prediction.

2.
Mitochondrion ; 2020 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-31923469

RESUMO

Beauveria bassiana, Cordyceps militaris and Ophiocordyceps sinensis (Ascomycotina) are traditional Chinese medicines. Here, mitogenomes of these three Ascomycotina fungi were sequenced and de-novo assembled using single-molecule real-time sequencing. The results showed that their complete mitogenomes were 31,258, 31,854 and 157,584 bp, respectively, with sequencing depth approximately 278,760×, 326,283× and 69,385×. Types of repeat sequences were mainly (AA)n, (AAT)n, (TA)n and (TATT)n. DNA methylation motifs were revealed in DNA modifications of these three fungi. We discovered new models of RNA editing through analysis of transcriptomes from B. bassiana and C. militaris. These data lay a solid foundation for further genetic and biological studies about these three fungi, especially for elucidating the mitogenome evolution and exploring the regulatory mechanism of adapting environment.

3.
Investig Clin Urol ; 61(1): 75-80, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31942466

RESUMO

Purpose: To examine associations if any between changes in voiding function, hematuria, and bladder ultrasonography metrics in murine cyclophosphamide-induced chemical cystitis. Materials and Methods: Cystitis was induced in 6 female mice by an intraperitoneal injection of cyclophosphamide (300 mg/kg). Voiding frequency, void volume, hematuria assessment, and ultrasonographic measurements of the bladder were obtained at baseline, days 1 to 5, and days 9, 11, and 13. Voiding was induced with preferred sweet drinking solution and voiding data collected using an automated data collection system in 135 minute sessions. Bladder wall thickness, lumen volume, and vascular Doppler were acquired using a high definition ultrasound system. Spearman's correlation was used to analyze the association between the voiding changes, hematuria, and ultrasound findings. Results: Hematuria was present 24 hours after cyclophosphamide injection. All animals displayed increased bladder vascularity, bladder wall thickness, and void frequency that was associated with concurrent decreased total and average void volumes. Increased bladder wall vascularity was correlated with the presence of hematuria (r=0.59, p<0.01) and bladder wall thickness (r=0.79, p<0.01). Hematuria correlated with increased void frequency (r=0.34, p<0.01). Average void volume was negatively correlated with hematuria (r=-0.50, p<0.01) and frequency (r=-0.38, p<0.01). Conclusions: High-definition ultrasound imaging permits in vivo monitoring of changes in bladder morphology associated with voiding function in relation to cyclophosphamide-induced cystitis. Ultrasound imaging of the bladder may assist in differential diagnosis of bladder dysfunction.

4.
Eur J Med Chem ; 187: 111936, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31855793

RESUMO

The use of antagonists for each adenosine receptor (AR) subtype as potent clinical candidates is of growing interest due to their involvement in the treatment of various diseases. The recent resolution of several A1 and A2A ARs X-ray structures provides opportunities for structure-based drug design. In this study, we describe the discovery of novel A1AR antagonists by applying a multistage virtual screening approach, which is based on random forest (RF), e-pharmacophore modeling and docking methods. A multistage virtual screening approach was applied to screen the ChemDiv library (1,492,362 compounds). Among the final hits, 22 compounds were selected for further radioligand binding assay analysis against human A1AR, and 18 compounds (81.82% success) exhibited nanomolar or low micromolar binding potency (Ki). Then, we selected six compounds (pKi > 6) to further evaluate their antagonist profile in a cAMP functional assay, and we found that they had low micromolar antagonistic activity (pIC50 = 5.51-6.38) for the A1AR. Particularly, four of six compounds (pKi > 6) showed very good affinity (pKi = 6.11-7.13) and selectively (>100-fold) for A1AR over A2AAR. Moreover, the novelty analysis suggested that four of six compounds (pKi > 6) were dissimilar to existing A1AR antagonists and hence represented novel A1AR antagonists. Further molecular docking and molecular dynamics (MD) studies showed that the three selective compounds 15, 20 and 22 were stabilized (RMSlig value ≤ 2 Å) inside the binding pocket of A1AR with similar orientations to the docking pose in 100-ns MD simulations, whereas they escaped from the binding area of A2AAR with larger values of RMSlig (RMSlig ≥ 2 Å). We hope that these findings provide new insights into the discovery of drugs targeting A1AR and facilitate research on new drugs and treatments for A1AR-related human pathologies.

5.
Indian J Ophthalmol ; 68(1): 216-218, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31856529

RESUMO

Congenital microphthalmia (CM) is a rare anomaly of the fetal orbit, results from developmental defects of the primary optic vesicle, and is characterized by a reduced eyeball volume and axial diameter. Fetal CM cases have rarely been reported. Herein, we present a case of two fetuses with bilateral CM from the same parents, diagnosed using ultrasonography (US) and magnetic resonance imaging (MRI). We found that the antepartum US and MRI measurements were smaller than the postpartum ones. Genetic testing of the parents and fetuses revealed that GL12 gene mutation may be associated with CM.

7.
Analyst ; 2019 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-31820748

RESUMO

Raman spectroscopy has been employed for studying the hydrogen bonding states of water molecules for decades, however, Raman imaging data contain thousands of spectra, making it challenging to obtain information on water with different hydrogen bonds. In the current study, a novel method combining confocal Raman microscopy (CRM) imaging with the iterative curve fitting algorithms was developed to determine the distribution of water contents at the cellular level and water states with different hydrogen bonds in apple tissues. Raman imaging data ranging from 2700 to 3800 cm-1 were acquired from whole cells in the apple tissue, which were then decomposed into seven sub-peaks using the fixed-position Gaussian iterative curve fitting (FPGICF) algorithm. The content and hydrogen bonding states of cellular water were calculated as the area sum of the OH stretching vibration and the area ratio of DA-OH over DDAA-OH stretching vibration or the number of hydrogen bonds of each water molecule, respectively. Finally, the area of each sub-peak, the area sum of the OH stretching vibration, and the area ratio of DA-OH over DDAA-OH stretching vibration were used to visualize the distribution of each sub-peak, water contents and water states with different hydrogen bonds, respectively. In addition, it was found that the number of hydrogen bonds of each water molecule could also be considered as a criterion to describe the hydrogen bond states of water in apple tissues. The availability of such information should provide new insights for future study of cellular water in other food materials.

8.
Cell Signal ; : 109501, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31837464

RESUMO

Inactivation of glutathione S-transferase pi 1 (GSTP1) via hypermethylation is an early and common event in prostate carcinogenesis. Functional inactivation of GSTP1 increases the susceptibility to oxidative stress and enhance progression risk of the prostatic carcinoma. In this study, we hypothesized that the Piwi-interacting RNA (piRNA) could be a sequence-recognition and guidance molecule for induction of promoter methylation of GSTP1 facilitating prostate carcinogenesis. We found that piR-31470 was highly expressed in prostate cancer cells, and piR-31470 could bind to piwi-like RNA-mediated gene silencing 4 (PIWIL4) to form the PIWIL4/piR-31470 complex. This complex could bind to the nascent RNA transcripts of GSTP1, and recruit DNA methyltransferase 1, DNA methyltransferase 3 alpha and methyl-CpG binding domain protein 2 to initiate and maintain the hypermethylation and inactivation of GSTP1. Our data demonstrated that the overexpression of piR-31470 inhibited the levels of GSTP1 and increased vulnerability to oxidative stress and DNA damage in human prostate epithelial RWPE1 cells. In conclusion, this study characterized the roles of the PIWIL4/piR-31470 complex in the regulation of the transcription of GSTP1 by methylating the CpG island of GSTP1. This discovery may provide a novel therapeutic strategy by targeting piRNAs for the epigenetic treatment of prostate cancer.

9.
Front Plant Sci ; 10: 1473, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31827478

RESUMO

Brassinosteroids (BRs) are a group of plant steroid hormones that play important roles in regulating plant development. In addition, BRs show considerable functional redundancy with other plant hormones such as gibberellins (GAs). BRASSINAZOLE RESISTANT1 (BZR1) and BRI1-EMS-SUPPRESSOR1 (BES1) transcription factors are negative feedback regulators of BR biosynthesis. This study provides evidence for the roles of MdBZR1 and MdBZR1-2like in promoting GA production. These results also show that BRs regulate GA biosynthesis to improve salt tolerance in apple calli. Moreover, this research proposes a regulatory model, in which MdBZR1 and MdBZR1-2like bind to the promoters of GA biosynthetic genes to regulate their expression in a BR-dependent manner. The expression of key GA biosynthetic genes, MdGA20ox1, MdGA20ox2, and MdGA3ox1 in yeast helps to maintain normal growth even under intense salt stress. In summary, this study underscores the roles of MdBZR1 and MdBZR1-2like in improving salt tolerance by regulating GA biosynthesis in apple calli.

10.
J Clin Transl Sci ; 3(6): 332-343, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31827907

RESUMO

Introduction: Recently, volumetric absorptive microsampling (VAMS) has been used for accurate sampling of a fixed peripheral blood volume (10 µL) on a volumetric swab, and long-term sample storage. The mPlex-Flu assay is a novel, high-throughput assay that simultaneously measures the concentration of antibodies against the hemagglutinin (HA) proteins from multiple influenza virus strains with ≤5 µL of serum. Here we describe combining these two methods to measure multidimensional anti-influenza IgG activity in whole blood samples collected by a finger stick and VAMS, with correction for serum volume based on simultaneous hemoglobin measurement. Methods: We compared capillary blood samples obtained from a finger stick using a VAMS device with serum samples collected by traditional phlebotomy from 20 subjects, with the influenza antibody profiles measured by the mPlex-Flu assay. Results: We found that results with the two sampling methods were highly correlated within subjects and across all influenza strains (mean R 2 = 0.9470). Adjustment for serum volume, based on hemaglobin measurement, was used to estimate serum volume of samples and improved the accuracy. IgG measurements were stable over 3 weeks when VAMS samples were stored at room temperature or transported using a variety of shipping methods. Additionally, when volunteers performed finger-stick VAMS at-home by themselves, the comparison results of anti-HA antibody concentrations were highly consistent with sampling performed by study personnel on-site (R 2 = 0.9496). Conclusions: This novel approach can provide a simple, accurate, and low-cost means for monitoring the IgG anti-influenza HA antibody responses in large population studies and clinical trials.

11.
Nicotine Tob Res ; 2019 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-31830263

RESUMO

INTRODUCTION: Electronic cigarette use (vaping) has increased in recent years. Chronic obstructive pulmonary disease (COPD) is the third leading cause of death associated with smoking. METHODS: Based on 2016 and 2017 Behavioral Risk Factor Surveillance System (BRFSS) national survey data on 891,242 adult participants who indicated their smoking and vaping status, the cross-sectional association of vaping with self-reported COPD diagnosis was investigated, using univariable and multivariable weighted logistic regression models. RESULTS: Compared to never users, while dual users showed the highest association with self-reported COPD diagnosis (aOR=4.39, 95% CI: 3.98, 4.85), current vapers who were either ex-smokers or never smoked showed significantly higher association with self-reported COPD diagnosis (aOR= 3.24, 95% CI: 2.78, 3.78 and aOR=1.47, 95% CI: 1.01, 2.12, respectively). Current vapers who were ex-smokers showed higher association with self-reported COPD diagnosis than ex-smokers who do not vape (aOR=1.27, 95% CI: 1.09, 1.48). Dual users showed higher association with self-reported COPD diagnosis than current smokers who do not vape (aOR=1.16, 95% CI: 1.05, 1.27). Ex-smokers showed significantly less association with self-reported COPD diagnosis (aOR=0.67, 95% CI: 0.64, 0.71) than current smokers. Current vapers who were either ex-smokers or never smoked had less association with self-reported COPD diagnosis compared to current smokers, with aOR=0.85 (95% CI: 0.73, 0.99) and aOR=0.39 (95% CI: 0.27, 0.56). CONCLUSIONS: Vaping is significantly associated with self-reported COPD diagnosis in adults, even among vapers who never smoked. Whether there is a benefit for COPD of switching from smoking to vaping requires study of the long-term effects of vaping.

12.
Biomed Pharmacother ; 123: 109616, 2019 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-31881485

RESUMO

Breast cancer (BC) is a major contributor of cancer-associated mortality in women. It is essential to find new therapeutic targets and drugs. Polyrhachis vicina Rogers is one of the Traditional Chinese Medicine (TCM). Our previous studies have shown an active fraction of Polyrhachis vicina Rogers (AFPR) has significant anti-inflammatory activity, suggesting its anti-cancer effect. Here, we aimed to explore the inhibitory effects of AFPR on BC and reveal its mechanism. The effects of AFPR on BC were examined by cell proliferation assay, wound healing assay, invasion assay and xenograft assay. Microarray sequencing, qRT-PCR, Western blot, chromatin immunoprecipitation assay and luciferase reporter assay were performed to investigate the regulation of AFPR on related genes and underlying mechanisms. As a result, AFPR suppressed BC cell growth, migration and invasion and inhibited tumor growth. LncRNA NKILA was most prominently upregulated in AFPR-treated MCF7 cells. AFPR inactivated NF-κB signaling pathway via regulating NKILA. Furthermore, AFPR regulated the expression of NKILA by inhibiting its transcript suppressor EGR1. This study firstly indicated that AFPR was a potential inhibitor of BC development via regulating EGR1/NKILA/NF-κB axis.

13.
Comput Biol Chem ; 84: 107194, 2019 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-31881526

RESUMO

The tumor suppressor p53, a transcription factor, plays a critical role in many cellular processes, including DNA repair and apoptosis and cell cycle arrest. Missense mutations in the p53 have closely related to human cancer. R249S mutation at the p53 core DNA binding domain (DBD) is frequently observed in hepatocellular carcinoma. This mutation is away from the p53 DBD-DNA binding interface. However, how the R249S mutation causes the structural changes of p53 DBD that lead to weak the binding of p53 mutant to DNA has not been clearly understood. Here, microsecond-scale molecular dynamics (MD) simulations of p53 DBD in the wild type (WT) and R249S mutated states in the absence of DNA binding were performed to explore the effect of the R249S mutation on the conformational dynamics of p53 DBD. The R249S mutation does not cause the global conformational changes, and it only affects the local domains at the mutation site and the DNA binding interface, particularly at the S1-S2 turn. The allosteric effects of the S1-S2 turn induced by the R249S mutation lead to the extension of the S1-S2 turn into the ß-strands, which in turn interferes with the binding of DNA at the major groove. The results can help decipher the allosteric regulatory mechanism by which the R249S mutation of p53 DBD affects the p53 DBD-DNA interactions.

14.
Sci Total Environ ; 707: 136139, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31863983

RESUMO

Di-n-butyl phthalate (DBP), as one of the environmental chemicals, can cause male reproductive decline including testicular hypoplasia and impairments of spermatogenesis. Testicular inflammation is positively related to decline in male reproductive function. However, whether exposure to DBP in utero can cause testicular inflammation in progeny has not been studied. In this study, we established an animal model and observed that DBP exposure during gestation induced testicular inflammation in progeny with the increased expression of pro-inflammatory cytokines and chemokines including tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), monocyte chemotactic protein-1 (MCP-1) and CXC chemokine ligand-10 (CXCL-10), representing the activation of the nuclear factor kappa B (NF-κB). However, NF-κB was activated within 1 h in Sertoli cells (SCs) when exposed to MBP (a metabolite of DBP) in vitro. Meanwhile, we detected increased expression of inflammatory NLR family pyrin domain containing 3 (NLRP3), resulting from Pellino2-mediated NLRP3 inflammasome priming. Further, we confirmed that the activation of the NLRP3/caspase-1/IL-1ß canonical inflammasome pathway induced secretion of inflammatory factors of SCs and immune response, and INF39 (an inhibitor of NLRP3) could inhibit the inflammation in vitro. Collectively, these findings indicated that NLRP3 inflammasomes played key roles in DBP-induced inflammation in testicular SCs.

15.
BMC Genomics ; 20(1): 987, 2019 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-31842754

RESUMO

BACKGROUND: As a photoperiod-sensitive and self-pollinated species, the growth periods traits play important roles in the adaptability and yield of soybean. To examine the genetic architecture of soybean growth periods, we performed a genome-wide association study (GWAS) using a panel of 278 soybean accessions and 34,710 single nucleotide polymorphisms (SNPs) with minor allele frequencies (MAF) higher than 0.04 detected by the specific-locus amplified fragment sequencing (SLAF-seq) with a 6.14-fold average sequencing depth. GWAS was conducted by a compressed mixed linear model (CMLM) involving in both relative kinship and population structure. RESULTS: GWAS revealed that 37 significant SNP peaks associated with soybean flowering time or other growth periods related traits including full bloom, beginning pod, full pod, beginning seed, and full seed in two or more environments at -log10(P) > 3.75 or -log10(P) > 4.44 were distributed on 14 chromosomes, including chromosome 1, 2, 3, 5, 6, 9, 11, 12, 13, 14, 15, 17, 18, 19. Fourteen SNPs were novel loci and 23 SNPs were located within known QTLs or 75 kb near the known SNPs. Five candidate genes (Glyma.05G101800, Glyma.11G140100, Glyma.11G142900, Glyma.19G099700, Glyma.19G100900) in a 90 kb genomic region of each side of four significant SNPs (Gm5_27111367, Gm11_10629613, Gm11_10950924, Gm19_34768458) based on the average LD decay were homologs of Arabidopsis flowering time genes of AT5G48385.1, AT3G46510.1, AT5G59780.3, AT1G28050.1, and AT3G26790.1. These genes encoding FRI (FRIGIDA), PUB13 (plant U-box 13), MYB59, CONSTANS, and FUS3 proteins respectively might play important roles in controlling soybean growth periods. CONCLUSIONS: This study identified putative SNP markers associated with soybean growth period traits, which could be used for the marker-assisted selection of soybean growth period traits. Furthermore, the possible candidate genes involved in the control of soybean flowering time were predicted.

16.
J Exp Bot ; 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31740930

RESUMO

The dormancy-associated MADS-box genes PpDAM5 and PpDAM6 have been shown to play an important role in bud endodormancy. However, the molecular regulatory mechanism of PpDAM5 and PpDAM6 involvement in peach endodormancy is still unclear. Here, through yeast one-hybrid (Y1H) screening, we isolated PpTCP20, a TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factor in the peach cultivar 'Zhongyou 4' (Prunus persica var. nectarina). PpTCP20 was localized in the nucleus and was capable of forming a homodimer. The electrophoretic mobility shift assay (EMSA) demonstrated that PpTCP20 binds to a GCCCR element in the promoter of PpDAM5 and PpDAM6, and transient dual luciferase (Dual-LUC) experiments showed that PpTCP20 inhibited PpDAM5 and PpDAM6 expression as the peach flower bud endodormancy release period approached. Furthermore, PpTCP20 interacted with PpABF2 to form heterodimers to regulate peach bud endodormancy, and the content of abscisic acid (ABA) decreased with the release of peach flower bud endodormancy. In addition, PpTCP20 inhibited PpABF2 expression to regulate peach flower bud endodormancy. Taken together, our results suggest that PpTCP20 regulates peach flower bud endodormancy by negatively regulating the expression of PpDAM5, PpDAM6 and interacting with PpABF2. Therefore, these results reveal the novel mechanism regulating bud endodormancy in perennial deciduous trees, including peach.

17.
Food Funct ; 10(12): 8310, 2019 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-31710079

RESUMO

Correction for 'Insights into melanoidin conversion into fluorescent nanoparticles in the Maillard reaction' by Dongmei Li et al., Food Funct., 2019, 10, 4414-4422.

18.
Reproduction ; 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31710595

RESUMO

Health risk of human exposure to microcystin-leucine arginine (MC-LR) has drawn more and more attention in recent years. In the present study, MC-LR inhibited miR-3473g expression of mouse granulosa cells both in vitro and in vivo. By dual-luciferase reporter assay, we confirmed miR-3473g is able to bind the 3'-untranslated region of steroidogenic acute regulatory protein (StAR) mRNA and suppress StAR expression. Thus, down-regulation of miR-3473g after MC-LR exposure led to StAR overexpression. Excessive StAR probably transported much more cholesterol into the inner membrane of the mitochondria and finally resulted in overproduction of progesterone. Our results revealed that MC-LR exposure was associated with premature luteinization of granulosa cells and may adversely affect women's fertility.

19.
Front Genet ; 10: 907, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31681403

RESUMO

Autism spectrum disorders (ASD) are hypothesized to originate in utero from perturbations in neural stem cell niche regions of the developing brain. Dynamic epigenetic processes including DNA methylation are integral to coordinating typical brain development. However, the extent and consequences of alterations to DNA methylation states in neural stem cell compartments in ASD are unknown. Here, we report significant DNA methylation defects in the subventricular zone of the lateral ventricles from postmortem brain of 17 autism diagnosed compared to 17 age- and gender-matched typically developing individuals. Both array- and sequencing-based genome-wide methylome analyses independently revealed that these alterations were preferentially targeted to intragenic and bivalently modified chromatin domains of genes predominately involved in neurodevelopment, which associated with aberrant precursor messenger RNA splicing events of ASD-relevant genes. Integrative analysis of our ASD and typically developing postmortem brain methylome datasets with that from fetal brain at different neurodevelopmental stages revealed that the methylation states of differentially methylated loci associated with ASD remarkably resemble the methylation states at earlier time points in fetal brain development. This observation was confirmed using additional methylome datasets from three other brain regions. Altogether, these findings implicate an epigenetic delay in the trajectory of normal DNA methylation states during the course of brain development that may consequently lead to deleterious transcriptomic events in ASD and support the hypothesis of an early developmental origin of ASD.

20.
J Extracell Vesicles ; 8(1): 1684816, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31762962

RESUMO

Extracellular vesicles (EVs) play a vital role in normal lung physiology to maintain homeostasis in the airways via intercellular communication. EVs include exosomes and microvesicles, and are characterized by their phospholipid bilayers. EVs have been recognized as novel circulating biomarkers of disease, which are released by different cell types. In this study, we used different EV isolation and purification methods to characterize the plasma-derived EV miRNAs from non-smokers, smokers and patients with COPD. A small RNA sequencing (RNA-seq) approach was adapted to identify novel circulating EV miRNAs. We found that plasma-derived EVs from non-smokers, smokers and patients with COPD vary in their size, concentration, distribution and phenotypic characteristics as confirmed by nanoparticle tracking analysis, transmission electron microscopy, and immunoblot analysis of EV surface markers. RNA-seq analysis confirmed the most abundant types of small RNAs, such as miRNAs, tRNAs, piRNAs snRNAs, snoRNAs and other biotypes in plasma-derived EVs. We mainly focused on miRNAs as novel biomarkers in smokers and patients with COPD for further analysis. Differential expression by DESeq2 identified distinct miRNA profiles (up-regulated: miR-22-3p, miR-99a-5p, miR-151a-5p, miR-320b, miR-320d; and down-regulated: miR-335-5p, miR-628-3p, miR-887-5p and miR-937-3p) in COPD versus smokers or non-smokers in a pairwise comparison. Gene set enrichment analysis (GSEA) of differentially expressed miRNAs revealed the top pathways, gene ontology and diseases associated with smokers and patients with COPD. We selectively validated miRNAs in EVs isolated from BEAS-2B cells treated with cigarette smoke extract by quantitative PCR analysis. For the first time, we report that plasma-derived EV miRNAs are novel circulating pulmonary disease biomarkers. Thus, molecular profiling of EV miRNAs has great translational potential for the development of biomarkers that may be used in the diagnosis, prognosis, and therapeutics of COPD.

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