MicroRNA
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MicroRNAs (miRNAs), a type of short noncoding RNA molecule (21-23 nucleotides), mediate repressive gene regulation through RNA silencing at the posttranscriptional level and play an important role in the defense response to abiotic and biotic stresses. miRNAs of the plant system have been studied in model crops for their diverse regulatory role while less is known about their significance in other plants whose genome and transcriptome data are scarce in the database, including eggplant (Solanum melongena L.). In the present study, a next-generation sequencing platform was used for the sequencing of miRNA, and real-time quantitative PCR for miRNAs was used to validate the gene expression patterns of miRNAs in Solanum melongena plantlets infected with the bacterial wilt-causing pathogen Ralstonia solanacearum (R. solanacearum). Sequence analyses showed the presence of 375 miRNAs belonging to 29 conserved families. The miR414 is highly conserved miRNA across the plant system while miR5658 and miR5021 were found exclusively in Arabidopsis thaliana surprisingly, these miRNAs were found in eggplants too. The most abundant families were miR5658 and miR414. Ppt-miR414, hvu-miR444b, stu-miR8020, and sly miR5303 were upregulated in Pusa purple long (PPL) (susceptible) at 48 h postinfection, followed by a decline after 96 h postinfection. A similar trend was obtained in ath-miR414, stu-mir5303h, alymiR847-5p, far-miR1134, ath-miR5021, ath-miR5658, osa-miR2873c, lja-miR7530, stu-miR7997c, and gra-miR8741 but at very low levels after infection in the susceptible variety, indicating their negative role in the suppression of host immunity. On the other hand, osa-miR2873c was found to be slightly increased after 96 hpi from 48 hpi. Most of the miRNAs under study showed relatively lower expression in the resistant variety Arka Nidhi after infection than in the susceptible variety. These results shed light on a deeper regulatory role of miRNAs and their targets in regulation of the plant response to bacterial infection. The present experiment and their results suggested that the higher expression of miRNA leads to a decline in host mRNA and thus shows susceptibility.
- Publikační typ
- časopisecké články MeSH
PURPOSE OF THE STUDY The aim of the study was to determine miR-146a-5p, miR-223-3p and miR-23a-3p by an enzyme immunoassay in patients with inflammatory and non-inflammatory joint effusion and to verify the usefulness of these miRNAs as biomarkers of joint inflammation. MATERIAL AND METHODS Synovial fluid (SF) samples were collected from 82 patients. The group consisted of 60 non-inflammatory, 11 inflammatory-non-pyogenic, 11 inflammatory-pyogenic SF. SF miRNA was isolated by RNA Isolation Kit Plasma/Serum. The concentrations of miRNA were determined by enzyme-linked immunosorbent assays (ELISA), C-reactive protein, interleukin-6 and procalcitonin on automatic analyser, presepsin on POCT system, interleukin-1 and human neutrofil defensins 1-3 by ELISA. RESULTS A statistically significant negative correlation was found between miR-146-5p and miR-223-3p, WBC, IL-1β, IL-6 and CRP (P < 0.05) in all groups; a statistically significant positive correlation was found between miR-223-3p and miR-23a-3p, WBC, PMN, IL-1beta, IL-6 and HNP1-3, as well as a positive correlation of miR-23a-3p with IL-1β, IL-6 and HNP1-3. A statistically significant difference was found between miR-146a-5p, miR-223-3p and miR-23a-3p and individual SF groups, P = 0.006, P < 0.001, respectively. PMN, WBC, Il-1β, IL-6, HNP 1-3 predicted the inflammatory processes with excellent diagnostic power (AUC > 0.9). The clinical relevance expressed by effect size was the strongest in miR-223-3p, PMN, IL-1 , HNP 1-3 between non-inflammatory and inflammatory-pyogenic group. CONCLUSIONS Our study quantified the SF miRNA by ELISA. We have shown that miR-146a-5p, miR-223-3p and miR-23a-3p can be an important group of biomarkers for the detection and monitoring of various pathophysiological conditions in synovial fluid, including inflammatory conditions. Key words: miRNA, synovial fluid, inflammatory joint disease, enzyme-linked immunosorbent assay.
- MeSH
- biologické markery MeSH
- lidé MeSH
- lipopolysacharidové receptory MeSH
- mikro RNA * genetika MeSH
- peptidové fragmenty MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- lipopolysacharidové receptory MeSH
- mikro RNA * MeSH
- peptidové fragmenty MeSH
- presepsin protein, human MeSH Prohlížeč
PURPOSE OF THE STUDY Nitinol (NiTi) is a biomaterial widely used in medicine based on super-elastic and shape memory properties. miR-124 has a key role in inflammatory process, osteoblasts differentiation, and mineralization. The aim of study was evaluating the differences in gene expression of miR-124 of human physiological osteoblasts (HOB) and human osteoarthritic osteoblasts (OSBA) as a response to NiTi alloy in different heat treatments. MATERIAL AND METHODS The cells were cultivated with NiTi discs with/without addition of bacterial lipopolysaccharide (LPS) for 72 hours. MicroRNAs were isolated, underwent reverse transcription and were analyzed by RT-PCR. RESULTS As a response to LPS, HOB overexpressed miR-124, while in OSBA expression change did not occur. Overexpression was also observed in both cell lines as a response to hydrogen and helium treated NiTi discs. HOB expressed significantly higher amount of miR-124 than OSBA as a response to hydrogen treatment of NiTi discs. In addition, hydrogen treatment caused significantly higher expression in HOB than LPS. The combination of NiTi disc and LPS treatment in HOB didn't cause any expression changes. Comparing to LPS-only treatment, the expression in HOB with combination of LPS and alloy was significantly lower. In OSBA, the expression was increased by the combination of LPS and hydrogen disc, in case of helium disc, the expression was decreased. CONCLUSIONS In conclusion, human physiological and osteoarthritic osteoblasts respond to NiTi alloy with both surface (hydrogen and helium atmosphere) treatment by overexpression of miR-124. The effect of LPS as inflammatory modulator suggests the presence of an "anti-inflammatory preconditioning" in osteoarthritic osteoblasts, as physiological osteoblasts overexpression was significantly higher. Key words: nitinol, osteoblast, miR-124, lipopolysaccharide.
- MeSH
- helium metabolismus farmakologie MeSH
- lidé MeSH
- lipopolysacharidy * farmakologie metabolismus MeSH
- mikro RNA * genetika metabolismus farmakologie MeSH
- osteoartróza genetika MeSH
- osteoblasty metabolismus MeSH
- slitiny metabolismus farmakologie MeSH
- titan MeSH
- vodík metabolismus farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- helium MeSH
- lipopolysacharidy * MeSH
- mikro RNA * MeSH
- MIRN124 microRNA, human MeSH Prohlížeč
- nitinol MeSH Prohlížeč
- slitiny MeSH
- titan MeSH
- vodík MeSH
PURPOSE OF THE STUDY Articular cartilage injury is a common disease in daily life, with a high incidence. The aim of this study was to investigate the effect and mechanism of miRNA-140-3p in bone mesenchymal stem cells (BMSCs)-derived exosomes under hypoxia on inflammatory articular chondrocytes. MATERIAL AND METHODS To simulate the pathological status of arthritis, rat chondrocytes were used to establish the osteoarthritis (OA) model by IL-1β (10 μg/ml) as a modulating in vitro, and exosomes were isolated by differential ultra-high speed centrifugation. The cell counting kit-8, wound healing and flow cytometry assays were utilized to assess proliferation, migration and apoptosis of chondrocytes, respectively. Lipogenic and chondrogenic differentiation of chondrocytes were detected by oil red O staining and toluidine blue staining individually. The expressions of miR-140-3p and chondrocyte-specific gene mRNA were investigated using qRT-PCR. Western blot was applied to assess chondrocyte associated proteins and BMSC-Exo surface protein markers, and immunohistochemistry was adopted to detect the staining of collagen I and II. RESULTS Under scanning electronic microscope, the shape of exosomes was almost round. Exosome treatment prominently impaired the inhibition of chondrocytes' proliferative and migrative ability by IL-1β. It was found hypoxia had a more marked impact on proliferation, expression of collagen II and apoptosis in OA chondrocytes than normoxia, as well as a stronger effect on weakening adipose differentiation and enhancing chondrogenic differentiation in inflammatory chondrocytes. Furthermore, incubation with BMSC-Exo overexpressing miR-140-3p can remarkably increase the survival rate and migration in inflammatory chondrocytes. In addition, overexpression of miR-140-3p was found to enhance the chondrogenic differentiation of inflammatory chondrocytes. Furthermore, we found that the healing effect of exosomes on inflammatory chondrocytes under hypoxic conditions was produced by a rise in miR-140-3p expression within them and that hypoxia-mediated upregulation of miR-140-3p expression occurred through HIF-1α. CONCLUSIONS Under hypoxia, BMSC-Exo enhanced the chondrogenic phenotype, increased the viability of inflammatory chondrocytes. The overexpression of miR-140-3p in BMSC-Exo is beneficial to protect joints and delaying the pathogenesis in OA. Key words: HIF-1α, apoptosis, lipogenic differentiation, chondrogenic differentiation.
- MeSH
- antiflogistika MeSH
- exozómy * genetika MeSH
- hypoxie MeSH
- krysa rodu Rattus MeSH
- mikro RNA * genetika MeSH
- osteoartróza * genetika terapie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antiflogistika MeSH
- mikro RNA * MeSH
- MIRN140 microRNA, rat MeSH Prohlížeč
OBJECTIVE: MicroRNAs (miRs) are noncoding, single-stranded regulatory RNA molecules involved in the posttranscriptional regulation of gene expression. They control the development and maintenance of the diverse cellular processes including proliferation, differentiation, motility and apoptosis. Expression of miRs is tissue-specific and each alteration of the tissue miR profile is associated with a distinct disease status. STUDY DESIGN: We reviewed the literature on the expression of miRs in thyroid tumors, focusing on methodology and diagnostic and prognostic output. Separately, we analyzed 11 studies on miR profiles in thyroid cytological material. RESULTS: Numerous studies have evaluated the miR profiles of thyroid tumors in an attempt to find a possible diagnostic and prognostic role. Both downregulation and upregulation of numerous miRs was found, but differences between the surgical pathology specimens and corresponding fine-needle aspirates in the expression of the same miRs were also reported. CONCLUSIONS: The results from surgically resected material cannot be extrapolated into preoperative use without validation. For diagnostic use, the strong overlap between follicular adenoma and follicular carcinoma miR profiles is challenging. In summary, miR-221 and miR-222 are consistently upregulated in different types of thyroid carcinomas and might be used as markers of malignancy.
- MeSH
- lidé MeSH
- mikro RNA genetika MeSH
- nádorové biomarkery genetika MeSH
- nádory štítné žlázy diagnóza genetika patologie chirurgie MeSH
- prediktivní hodnota testů MeSH
- regulace genové exprese u nádorů MeSH
- stanovení celkové genové exprese MeSH
- štítná žláza chemie patologie chirurgie MeSH
- tenkojehlová biopsie * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- mikro RNA MeSH
- nádorové biomarkery MeSH
Non-Alcoholic Fatty Liver Disease (NAFLD) is one of the most important causes of liver disease worldwide leading the foreground cause of liver transplantation. Recently miRNAs, small non-coding molecules were identified as an important player in the negative translational regulation of many protein-coding genes involved in hepatic metabolism. Visceral adipose tissue was found to take part in lipid and glucose metabolism and to release many inflammatory mediators that may contribute to progression of NAFLD from simple steatosis to Non-Alcoholic SteatoHepatitis. Since visceral adipose tissue enlargement and dysregulated levels of miRNAs were observed in patients with NAFLD, the aim of this paper is to reflect the current knowledge of the role of miRNAs released from visceral adipose tissue and NAFLD.
- Klíčová slova
- NAFLD, NASH, adipose tissue, inflammation, lipid metabolism, miRNA, microRNA, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis,
- MeSH
- lidé MeSH
- mikro RNA metabolismus MeSH
- nealkoholová steatóza jater metabolismus MeSH
- nitrobřišní tuk metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- mikro RNA MeSH
Pregnane X receptor (PXR) is the major regulator of xenobiotic metabolism. PXR itself is controlled by various signaling molecules including glucocorticoids. Moreover, negative feed-back regulation has been proposed at the transcriptional level. We examined the involvement of the 3'-untranslated region (3'-UTR) of NR1I2 mRNA and microRNAs in PXR- and glucocorticoid receptor (GR)-mediated regulation of NR1I2 gene expression. PXR ligands were found to significantly downregulate NR1I2 mRNA expression in a set of 14 human hepatocyte cultures. Similarly, PXR was downregulated by PCN in the C57/BL6 mice liver. In mechanistic studies with the full-length 3'-UTR cloned into luciferase reporter or expression vectors, we showed that the 3'-UTR reduces PXR expression. From the miRNAs tested, miR-18a-5p inhibited both NR1I2 expression and CYP3A4 gene induction. Importantly, we observed significant upregulation of miR-18a-5p expression 6 h after treatment with the PXR ligand rifampicin, which indicates a putative mechanism underlying NR1I2 negative feed-back regulation in hepatic cells. Additionally, glucocorticoids upregulated NR1I2 expression not only through the promoter region but also via 3'-UTR regulation, which likely involves downregulation of miR-18a-5p. We conclude that miR-18a-5p is involved in the down-regulation of NR1I2 expression by its ligands and in the upregulation of NR1I2 mRNA expression by glucocorticoids in hepatic cells.
- Klíčová slova
- 3′-UTR, 3′-untranslated region, CAR, constitutive androstane receptor, CYP3A4, cytochrome P450 3A4, Cytochrome P450 3A4, DEX, dexamethasone, DMEs, drug metabolizing enzymes, DMSO, dimethyl sulfoxide, ER, estrogen receptor, GRα, glucocorticoid receptor α, Gene expression, Gluc, Gaussia luciferase, Glucocorticoid, LBD, ligand binding domain, MRE, miRNA-response element, MicroRNA, NR, nuclear receptor, PB, phenobarbital, PCN, pregnenolone 16α-carbonitrile, PHHs, primary human hepatocytes, PPARα, peroxisome proliferator-activated receptor α, PXR, pregnane X receptor, Pregnane X receptor, RXRα, retinoid X receptor α, Regulation, Rif, rifampicin, SEAP, secreted alkaline phosphatase, miRNA, microRNA,
- Publikační typ
- časopisecké články MeSH
MicroRNAs (miRNAs, miRs) represent a group of powerful and versatile posttranscriptional regulators of gene expression being involved in the fine control of a plethora of physiological and pathological processes. Besides their well-established crucial roles in the regulation of cell cycle, embryogenesis or tumorigenesis, these tiny molecules have also been shown to participate in the regulation of lipid metabolism. In particular, miRs orchestrate cholesterol and fatty acids synthesis, transport, and degradation and low-density and high-density lipoprotein (LDL and HDL) formation. It is thus not surprising that they have also been reported to affect the development and progression of several lipid metabolism-related disorders including liver steatosis and atherosclerosis. Mounting evidence suggests that miRs might represent important "posttranscriptional hubs" of lipid metabolism, which means that one miR usually targets 3'-untranslated regions of various mRNAs that are involved in different steps of one precise metabolic/signaling pathway, e.g., one miR targets mRNAs of enzymes important for cholesterol synthesis, degradation, and transport. Therefore, changes in the levels of one key miR affect various steps of one pathway, which is thereby promoted or inhibited. This makes miRs potent future diagnostic and even therapeutic tools for personalized medicine. Within this chapter, the most prominent microRNAs involved in lipid metabolism, e.g., miR-27a/b, miR-33/33*, miR-122, miR-144, or miR-223, and their intracellular and extracellular functions will be extensively discussed, in particular focusing on their mechanistic role in the pathophysiology of atherosclerosis. Special emphasis will be given on miR-122, the first microRNA currently in clinical trials for the treatment of hepatitis C and on miR-223, the most abundant miR in lipoprotein particles.
- Klíčová slova
- Atherosclerosis, Cholesterol, HDL, LDL, Lipid metabolism, miR-122, miR-223, miR-33,
- MeSH
- ateroskleróza genetika metabolismus patologie MeSH
- cholesterol genetika metabolismus MeSH
- energetický metabolismus MeSH
- lidé MeSH
- lipoproteiny HDL genetika metabolismus MeSH
- lipoproteiny LDL genetika metabolismus MeSH
- metabolické sítě a dráhy MeSH
- metabolismus lipidů * MeSH
- mikro RNA genetika metabolismus MeSH
- regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- cholesterol MeSH
- lipoproteiny HDL MeSH
- lipoproteiny LDL MeSH
- mikro RNA MeSH
- MIRN122 microRNA, human MeSH Prohlížeč
- MIRN223 microRNA, human MeSH Prohlížeč
- MIRN33a microRNA, human MeSH Prohlížeč
Cardiovascular diseases are major causes of morbidity and mortality in developed countries. Cerebrovascular diseases, especially stroke, represent major burden of disability and economy impact. Major advances in primary and secondary prevention and therapy are needed in order to tackle this public health problem. Our better understanding of pathophysiology is essential in order to develop novel diagnostic and therapeutic tools and strategies. microRNAs are a family of important post-transcriptional regulators of gene expression and their involvement in the pathophysiology of cerebrovascular diseases has already been reported. Moreover, microRNAs may represent above-mentioned potential diagnostic and therapeutic tools in clinical practice. Within this chapter, we briefly describe basic epidemiology, aetiology and clinical manifestation of following cerebrovascular diseases: extracranial carotid atherosclerosis, acute stroke, intracranial aneurysms and cerebral arterio-venous malformations. Further, in each chapter, the current knowledge about the involvement of specific microRNAs and their potential use in clinical practice will be summarized. More specifically, within the subchapter "miRNAs in carotid atherosclerosis", general information about miRNA involvement in atherosclerosis will be described (miR-126, miR-17-92, miR-155 and others) with special emphasis put on miRNAs affecting carotid plaque progression and stability (e.g. miR-145, miR-146 or miR-217). In the subchapter "miRNAs in acute stroke", we will provide insight into recent knowledge from animal and human studies concerning miRNA profiling in acute stroke and their expression dynamics in brain tissue and extracellular fluids (roles of, e.g. let-7 family, miR-21, miR-29 family, miR-124, miR-145, miR-181 family, miR-210 and miR-223). Subchapters dealing with "miRNAs and AV malformations" and "miRNAs and intracranial aneurysms" will focus on miR-21, miR-26, miR-29 family and miR-143/145.
- Klíčová slova
- Carotid atherosclerosis, Cerebral arterio-venous malformations, Intracranial aneurysms, Ischemic stroke, microRNA,
- MeSH
- cerebrovaskulární poruchy diagnóza genetika terapie MeSH
- cévní mozková příhoda etiologie genetika terapie MeSH
- ischemie mozku komplikace MeSH
- lidé MeSH
- mikro RNA genetika MeSH
- modely nemocí na zvířatech MeSH
- mozek metabolismus patologie MeSH
- nemoci arterie carotis diagnóza genetika terapie MeSH
- regulace genové exprese * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- mikro RNA MeSH
MicroRNAs (miRNAs) in urine are examined as potential biomarkers. We examined the urine samples from 70 individuals (45 males, 25 females, mean age 65 years, range 20-84 years). Of the urine donors, 15 were healthy volunteers, 5 were patients with non-cancer diseases, 50 were patients with different stages of bladder cancer. To examine the spectrum of miRNAs in the cell-free fraction of urine, TaqMan Human miRNA Array Card A v.2.1 was used. A set of 30 miRNAs were found that are constantly present in urine supernatants independently of sex, age and health status of the subjects. We compared this set with miRNAs found in plasma, expressed in kidney and genito-urinary tract. Our results indicate that some miRNA could be transferred from the circulation into urine.
- Klíčová slova
- Genito-urinary tract, MicroRNA, Plasma, Transrenal passage of microRNAs, Urine supernatant,
- MeSH
- dospělí MeSH
- ledviny metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikro RNA krev genetika moč MeSH
- mladý dospělý MeSH
- nádorové biomarkery krev genetika moč MeSH
- nádory močového měchýře diagnóza genetika MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- regulace genové exprese u nádorů MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- mikro RNA MeSH
- nádorové biomarkery MeSH