RESUMEN

Existing delivery methods for RNAi therapeutics encounter challenges, including stability, specificity, and off-target effects, which restrict their clinical effectiveness. In this study, a novel miR-133a zipper nanoparticle (NP) system that integrates miRNA zipper technology with rolling circle transcription (RCT) to achieve targeted delivery and specific regulation of miR-133a in adipocytes, is presented. This innovative approach can greatly enhance the delivery and release of miR-133a zippers, increasing the expression of thermogenic genes and mitochondrial biogenesis. he miR-133a zipper NP is utilized for the delivery of miRNA zipper-blocking miR-133a, an endogenous inhibitor of Prdm16 expression, to enhance the thermogenic activity of adipocytes by modulating their transcriptional program. Inhibition of miR-133a through the miR-133a zipper NP leads to more significant upregulation of thermogenic gene expression (Prdm16 and Ucp1) than with the free miR-133a zipper strand. Furthermore, miR-133a zipper NPs increase the number of mitochondria and induce heat production, reducing the size of 3D adipose spheroids. In short, this study emphasizes the role of RNA NPs in improving RNAi stability and specificity and paves the way for broader applications in gene therapy. Moreover, this research represents a significant advancement in RNAi-based treatments, pointing toward a promising direction for future therapeutic strategies.

Asunto(s)

Adipocitos , MicroARNs , Nanopartículas , Termogénesis , Factores de Transcripción , MicroARNs/genética , MicroARNs/metabolismo , Nanopartículas/química , Termogénesis/genética , Animales , Ratones , Adipocitos/metabolismo , Adipocitos/citología , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Humanos , Células 3T3-L1 , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Mitocondrias/metabolismo

RESUMEN

BACKGROUND: Diabetic cardiomyopathy (DCM) is a serious health-threatening complication of diabetes mellitus characterized by myocardial fibrosis and abnormal cardiac function. Human umbilical cord mesenchymal stromal cells (hUC-MSCs) are a potential therapeutic tool for DCM and myocardial fibrosis via mechanisms such as the regulation of microRNA (miRNA) expression and inflammation. It remains unclear, however, whether hUC-MSC therapy has beneficial effects on cardiac function following different durations of diabetes and which mechanistic aspects of DCM are modulated by hUC-MSC administration at different stages of its development. This study aimed to investigate the therapeutic effects of intravenous administration of hUC-MSCs on DCM following different durations of hyperglycemia in an experimental male model of diabetes and to determine the effects on expression of candidate miRNAs, target mRNA and inflammatory mediators. METHODS: A male mouse model of diabetes was induced by multiple low-dose streptozotocin injections. The effects on severity of DCM of intravenous injections of hUC-MSCs and saline two weeks previously were compared at 10 and 18 weeks after diabetes induction. At both time-points, biochemical assays, echocardiography, histopathology, polymerase chain reaction (PCR), immunohistochemistry and enzyme-linked immunosorbent assays (ELISA) were used to analyze blood glucose, body weight, cardiac structure and function, degree of myocardial fibrosis and expression of fibrosis-related mRNA, miRNA and inflammatory mediators. RESULTS: Saline-treated diabetic male mice had impaired cardiac function and increased cardiac fibrosis after 10 and 18 weeks of diabetes. At both time-points, cardiac dysfunction and fibrosis were improved in hUC-MSC-treated mice. Pro-fibrotic indicators (α-SMA, collagen I, collagen III, Smad3, Smad4) were reduced and anti-fibrotic mediators (FGF-1, miRNA-133a) were increased in hearts of diabetic animals receiving hUC-MSCs compared to saline. Increased blood levels of pro-inflammatory cytokines (IL-6, TNF, IL-1ß) and increased cardiac expression of IL-6 were also observed in saline-treated mice and were reduced by hUC-MSCs at both time-points, but to a lesser degree at 18 weeks. CONCLUSION: Intravenous injection of hUC-MSCs ameliorated key functional and structural features of DCM in male mice with diabetes of shorter and longer duration. Mechanistically, these effects were associated with restoration of intra-myocardial expression of miRNA-133a and its target mRNA COL1AI as well as suppression of systemic and localized inflammatory mediators.

Asunto(s)

Diabetes Mellitus Experimental , Cardiomiopatías Diabéticas , Fibrosis , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , MicroARNs , Miocardio , Cordón Umbilical , Animales , Humanos , Masculino , Ratones , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Experimental/metabolismo , Cardiomiopatías Diabéticas/terapia , Cardiomiopatías Diabéticas/metabolismo , Cardiomiopatías Diabéticas/patología , Cardiomiopatías Diabéticas/genética , Fibrosis/terapia , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/metabolismo , Ratones Endogámicos C57BL , MicroARNs/genética , MicroARNs/metabolismo , Miocardio/metabolismo , Miocardio/patología , Cordón Umbilical/citología , Cordón Umbilical/metabolismo

RESUMEN

Objective To explore the correlation between the serum levels expression of microRNA(miR)-133a-3p,protein tyrosine phosphatase nonreceptor type 22(PTPN22)and the severity of psoriasis vulgaris.Methods A total of 86 patients with psoriasis vulgaris who were admitted to Cangzhou People's Hospital from January 2022 to June 2022 were collected as the observation group.They were separated into a progressive group(n=41)and a quiescent group(n=45)based on the area and severity of the skin lesions.Meantime,86 healthy individuals undergoing plastic surgery examinations were regarded as the control group.Real time fluorescent quantitative PCR(qRT-PCR)method was applied to detect the relative expression levels of miR-133a-3p and PTPN22 in serum.Target Scan Human website was applied to predict the targeting relationship between PTPN22 and miR-133a-3p.Spearman method was applied to analyze the correlation between the expression levels of miR-133a-3p and PTPN22 in serum of patients with psoriasis vulgaris,the psoriasis area and the psoriasis area and severity index score(PASI).Logistic regression was applied to analyze the influencing factors of severity in patients with psoriasis vulgaris.Results Compared with the control group,the serum miR-133a-3p(1.85±0.46 vs 1.05±0.21)expression level in the observation group was increased,while the PTPN22 mRNA(0.76±0.13 vs 1.02±0.18)expression level was reduced,and the difference were statistically significant(t=14.671,10.859,all P<0.05).Compared with the quiescent group,the serum miR-133a-3p(2.05±0.52 vs 1.67±0.41)expression level in the progressive group was increased,while the PTPN22 mRNA(0.66±0.11 vs 0.85±0.15)expression level was reduced and the differences were statistically significant(t=3.780,6.643,all P<0.05).Target Scan Human website predicted that there may be a targeting relationship between miR-133a-3p and PTPN22.Spearman analysis showed that there was a positive correlation between serum miR-133a-3p and PASI score in patients with psoriasis vulgaris(r=0.469,P<0.05),while serum TPN22 mRNA level was negatively correlated with PASI score(r=0.469,P<0.05).Serum miR-133a-3p[OR(95%CI)=2.884(1.261～6.595)]was an independent risk factor for the severity of psoriasis vulgaris,while PTPN22[OR(95%CI)=0.562(0.367～0.860)]was an independent protective factor(all P<0.05).Conclusion The expression level of miR-133a-3p in serum of patients with psoriasis vulgaris was increased,while the expression level of PTPN22 was reduced.The two were closely related to the PASI score and may to some extent reflect the severity of psoriasis patients.

RESUMEN

BACKGROUND: Previous cross-sectional studies have shown that several circulating microRNA levels are associated with hypertension, but there are no prospective studies among general populations. OBJECTIVE: We evaluated the impact of circulating inflammatory- and oxidative stress-responsive microRNAs on changes in blood pressure and the development of hypertension in normotensive Japanese. METHOD: The study subjects were 84 normotensive participants (33 men and 51 women) who were given a health examination in both 2012 and 2017. In five years, 29 participants developed hypertension. Serum levels of miRNAs (miR-21, miR-27a, and miR-133a) were measured using qRT-PCR. Odds ratios (ORs) and 95% confidence intervals (CIs) for incident hypertension were estimated by logistic regression analysis. RESULTS: Serum miR-27a and -133a levels were lower in newly hypertensive subjects compared with normotensive subjects. With 1-unit lower serum miR-27a and -133a, the confounders adjusted ORs and 95% CI for incident hypertension were 0.84 (0.72-0.96) and 0.75 (0.58-0.91), respectively. The group with high levels of serum miR-27a and -133a had lower ORs than the group with low levels of these miRNAs (OR and 95% CI of miR-27a: 0.29, 0.08-0.91; miR-133a: 0.08, 0.01-0.37, respectively). CONCLUSIONS: Circulating miR-27a and -133a are potential biomarkers for the prediction and prevention of hypertension.

Asunto(s)

MicroARN Circulante , Hipertensión , MicroARNs , Biomarcadores , Presión Sanguínea , MicroARN Circulante/genética , Femenino , Humanos , Hipertensión/diagnóstico , Hipertensión/epidemiología , Hipertensión/genética , Masculino , MicroARNs/genética

RESUMEN

The present study aimed to develop fast melting tablets (FMTs) using silymarin (SM) owing to FMTs rapid disintegration and dissolution. FMTs represent a pathway to help patients to increase their compliance level of treatment via facile administration without water or chewing beside reduction cost. One of the methods for FMTs formulation is lyophilization. Optimization of SM-FMTs was developed via a 32 factorial design. All prepared SM-FMTs were evaluated for weight variation, thickness, breaking force, friability, content uniformity, disintegration time (DT), and % SM released. The optimized FMT formula was selected based on the criteria of scoring the fastest DT and highest % SM released after 10 min (Q10). Optimized FMT was subjected to Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffraction (XRD), and scanning electron microscopy (SEM) besides investigating its lung-protective efficacy. All SM-FMT tablets showed acceptable properties within the pharmacopeial standards. Optimized FMT (F7) scored a DT of 12.5 ± 0.64 Sec and % SM released at Q10 of 82.69 ± 2.88%. No incompatibilities were found between SM and excipients, it showed a porous structure under SEM. The optimized formula decreased cytokines, up-regulated miRNA133a, and down-regulated miRNA-155 and COX-2 involved in the protection against lung toxicity prompted by HgCl2 in a manner comparable to free SM at the same dosage.

Asunto(s)

MicroARNs , Silimarina , Administración Oral , Animales , Humanos , Pulmón , Ratas , Silimarina/química , Silimarina/farmacología , Solubilidad , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Comprimidos/química

RESUMEN

Cadmium is a common environmental heavy metal pollutant that can accumulate over long periods of time and cause disease. Thus, analysis of the molecular mechanisms affected by cadmium in the body could be of great significance for the prevention and treatment of cadmium-related diseases. In this study, flow cytometry, immunofluorescence, transmission electron microscopy, H&E (Hematoxylin Eosin) staining and TUNEL (TdT-mediated dUTP Nick-End Labeling) assays were used to verify that cadmium induced apoptosis and immune responses in bovine mammary epithelial cells (BMECs) and in mouse mammary gland. Isolated BMECs cultured with or without cadmium were collected to screen miRNA (microRNA) using high-throughput sequencing. There were 42 differentially-expressed miRNAs among which 27 were upregulated and 15 downregulated including bta-miR-133a, bta-miR-23b-5p, bta-miR-29e, bta-miR-365-5p, bta-miR-615, bta-miR-7, bta-miR-11975, bta-miR-127, and bta-miR-411a. Among those, miR-133a (which can specifically target TGFB2 (Recombinant Transforming Growth Factor Beta 2) was the most significantly downregulated with a fold-change of 5.27 in BMECs cultured with cadmium. Application of the double luciferase reporter system, western blotting, and qRT-PCR (Quantitative Real-time PCR) revealed that circ08409 can directly bind to miR-133a. Experiments demonstrated that circRNA-08409 could adsorb bta-miR-133a. Both circ08409 and TGFB2 significantly increased apoptosis and altered expression level of a series of inflammatory factors in BMECs. In contrast, miR-133a decreased significantly apoptosis and inflammation in the cells. Compared with cultures receiving only cadmium, the miR-133a+cadmium cultures exhibited significant reductions in the occurrence of late apoptosis. Overall, results indicated that circ08409 could relieve the inhibitory effect of miR-133a on TGFB2 expression by combining with miR-133a and subsequently modulating cell proliferation, apoptosis and inflammation. Overall, the data suggested that the circ08409/miR-133a/TGFB2 axis might play a role in mediating the effect of cadmium on BMECs. As such, data provide novel insights into controlling hazards that cadmium could induce in the mammary gland.

Asunto(s)

Cadmio , MicroARNs , Animales , Apoptosis , Cadmio/toxicidad , Bovinos , Células Epiteliales , Inflamación/inducido químicamente , Ratones , MicroARNs/genética

RESUMEN

IMPACT STATEMENT: This article describes a method for producing microRNA (miRNA)-enriched extracellular vesicles in large quantities. It enables in vivo delivery of specific miRNA for therapeutic applications.

Asunto(s)

Vesículas Extracelulares/metabolismo , Técnicas de Transferencia de Gen , MicroARNs/administración & dosificación , MicroARNs/aislamiento & purificación , Animales , Femenino , Células HEK293 , Humanos , Ratones , Ratones Endogámicos C57BL , MicroARNs/sangre , MicroARNs/genética

RESUMEN

OBJECTIVE: To investigate the effects of hydrogen sulfide (H2S) on the negatively regulation of cardiomyocyte hypertrophy and the relationship between the effect of H2S with miRNA-133a-mediated Ca2+/calcineurin/NFATc4 signal pathway. METHODS: Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (Leica). The expression of brain natriuretic peptide(BNP), ß-myosin heavy chain(ß-MHC), cystathionase (CSE), miRNA-133a, calcineurin (CaN) were detected by qRT-PCR. The protein expressions of CaN、nuclear factors of activated T cells (NFATc4) were detected by Western blot. The concentration of H2S in the cardiomyocyte was detected by Elisa. The concentration of intracellular calcium was measured by calcium imaging using confocal microscope. The nuclear translocation of NFATc4 was checked by immuno-fluorescence cell staining technique. RESULTS: ①The level of system of CSE/H2S and expression of miRNA-133a were significantly reduced in cardiomyocyte hypertrophy. Pretreatment with NaHS increased the concentration of H2S and the expression of miRNA-133a mRNA in cardiomyocytes, and suppressed cardiomyocyte hypertrophy. ②The concentration of intracellular calcium, the expression of CaN and nulear protein NFATc4 were significantly increased, and the nuclear translocation of NFATc4 were obviously enhanced in cardiomyocyte hypertrophy. NaHS pretreatment markedly inhibited these effects of ISO induced cardiomyocyte hypertrophy. ③Application of antagomir-133a reversed the inhibitory effects of NaHS on cardiomyocyte hypertrophy, and increased the influx of intracellular calcium, and elevated the expression of CaN and nuclear protein NFATc4, and enhanced the nuclear translocation of NFATc4. CONCLUSIONS: H2S can negatively regulate cardiomyocyte hypertrophy. The effects might be associated with H2S increasing expression of miRNA-133a and inhibiting inactivation of Ca2+/calcineurin/NFATc4 signal pathway.

Asunto(s)

Calcineurina/metabolismo , Cardiomegalia/metabolismo , Sulfuro de Hidrógeno/metabolismo , MicroARNs/metabolismo , Factores de Transcripción NFATC/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Transducción de Señal , Animales , Cardiomegalia/inducido químicamente , Células Cultivadas , Cistationina gamma-Liasa/metabolismo , Miocitos Cardíacos/metabolismo , Cadenas Pesadas de Miosina/metabolismo , Péptido Natriurético Encefálico/metabolismo , Ratas

RESUMEN

Recent studies have recognized the involvement of microRNAs (miRNAs) in the development of osteoporosis, which regulate the balance between osteogenesis and osteoclasis. In this study, we investigated the regulation by miRNA-133a-5p on the osteoblast differentiation-associated markers in the mouse osteoblast-like MC3T3-E1 cells by RUNX2. First, we manipulated the miRNA-133a level in the MC3T3-E1 cells with 20 or 40 nM miR-133a-5p mimics, miR-133a-5p inhibitor, or scramble miRNA. Then, we quantified with real-time polymerase chain reaction (qRT-PCR) the expression of Collagen I, osteocalcin (OCN), and osteopontin (OPN) in the miR-133a-5p-manipulated MC3T3-E1 cells. And the confocal microscopy was also utilized to confirm the regulation by miR-133a-5p on the expression of the three molecules. We also investigated the extracellular matrix (ECM) mineralization and the alkaline phosphatase (ALP) activity in the miR-133a-5p-manipulated MC3T3-E1 cells. In addition, we explored the possible targeting by miR-133a-5p on RUNX2, which was a well-recognized promoter to osteoblast differentiation, with luciferase reporter, qRT-PCR, and Western blotting assay. Results demonstrated that the miRNA-133a-5p mimics markedly reduced, whereas the miRNA-133a-5p inhibitor significantly promoted the expression of Collagen I, OCN, and OPN, the ECM mineralization, and the ALP activity in MC3T3-E1 cells. The alignment analysis demonstrated a high homology between miRNA-133a-5p and the 3' UTR of RUNX2. Moreover, the luciferase reporter assay demonstrated that miRNA-133a-5p targeted the 3' UTR of RUNX2, and inhibited the expression of RUNX2 in both mRNA and protein levels. In conclusion, we identified the inhibition by miRNA-133a-5p to the expression of osteoblast differentiation markers, to the ECM mineralization, and to the ALP activity in MC3T3-E1 cells, by targeting the 3' UTR of RUNX2. Our study suggests that miRNA-133a-5p might be an important target to inhibit osteoblast differentiation in osteoporosis.

Asunto(s)

Diferenciación Celular , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , MicroARNs/genética , Osteoblastos/fisiología , Regiones no Traducidas 3' , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/metabolismo , Animales , Secuencia de Bases , Sitios de Unión , Biomarcadores/metabolismo , Calcificación Fisiológica , Línea Celular , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Expresión Génica , Ratones , Osteocalcina/genética , Osteocalcina/metabolismo , Osteopontina/genética , Osteopontina/metabolismo , Osteoporosis/metabolismo , Interferencia de ARN

RESUMEN

BACKGROUND: Corticosteroids are used in the treatment of asthma. The aim of this study was to determine the efficacy of anti-IgE and anti-TNF alpha as asthma treatments. METHODS: A mouse model of chronic asthma was developed. The fluticasone group was exposed to fluticasone and the anti-IgE and anti-TNF groups were administered anti-IgE or anti-TNF. IL-4, and IgE levels were measured, and histological analysis, pathological analysis and miRNA-126, miRNA-133a analyses were applied. RESULTS: The cell concentration in the BAL fluid decreased in all the treatment groups. The rate of perivascular and peribronchial cell infiltration decreased in the lung in the high-dose anti-IgE and anti-TNF groups. Smooth muscle thickness decreased in the lung tissue in the low-dose anti-IgE and anti-TNF groups. Bronchial wall thickness decreased in the lung tissue in the fluticasone+anti-IgE group. The IL-4 level in BAL fluid decreased in the high-dose anti-IgE, fluticasone+anti-IgE and anti-TNF groups. IgE levels increased in the BAL fluid in the high-dose anti-IgE and anti-TNF groups. The lymphocyte level increased in the BAL fluid in the high-dose anti-IgE group. The macrophage level decreased in the BAL fluid in the anti-TNF group. The relative expression of miRNA-126 increased in all groups. The relative expression of miRNA-133a decreased in the placebo and fluticasone groups. The relative expression of miRNA-133a increased in the low-dose anti-IgE, high-dose anti-IgE, fluticasone+anti-IgE and anti-TNF groups. CONCLUSIONS: The results showed that anti-IgE is successful as a treatment. Fluticasone+anti-IgE and anti-TNF were seen to be superior to other therapeutic modalities when used for prophylaxis.

Asunto(s)

Antiasmáticos/farmacología , Anticuerpos Antiidiotipos/farmacología , Asma/inmunología , Fluticasona/farmacología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Animales , Enfermedad Crónica , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos BALB C

RESUMEN

OBJECTIVE@#To investigate the effects of hydrogen sulfide (HS) on the negatively regulation of cardiomyocyte hypertrophy and the relationship between the effect of HS with miRNA-133a-mediated Ca/calcineurin/NFATc4 signal pathway.@*METHODS@#Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (Leica). The expression of brain natriuretic peptide(BNP), β-myosin heavy chain(β-MHC), cystathionase (CSE), miRNA-133a, calcineurin (CaN) were detected by qRT-PCR. The protein expressions of CaN、nuclear factors of activated T cells (NFATc4) were detected by Western blot. The concentration of HS in the cardiomyocyte was detected by Elisa. The concentration of intracellular calcium was measured by calcium imaging using confocal microscope. The nuclear translocation of NFATc4 was checked by immuno-fluorescence cell staining technique.@*RESULTS@#①The level of system of CSE/HS and expression of miRNA-133a were significantly reduced in cardiomyocyte hypertrophy. Pretreatment with NaHS increased the concentration of HS and the expression of miRNA-133a mRNA in cardiomyocytes, and suppressed cardiomyocyte hypertrophy. ②The concentration of intracellular calcium, the expression of CaN and nulear protein NFATc4 were significantly increased, and the nuclear translocation of NFATc4 were obviously enhanced in cardiomyocyte hypertrophy. NaHS pretreatment markedly inhibited these effects of ISO induced cardiomyocyte hypertrophy. ③Application of antagomir-133a reversed the inhibitory effects of NaHS on cardiomyocyte hypertrophy, and increased the influx of intracellular calcium, and elevated the expression of CaN and nuclear protein NFATc4, and enhanced the nuclear translocation of NFATc4.@*CONCLUSIONS@#HS can negatively regulate cardiomyocyte hypertrophy. The effects might be associated with HS increasing expression of miRNA-133a and inhibiting inactivation of Ca/calcineurin/NFATc4 signal pathway.

Asunto(s)

Animales , Ratas , Calcineurina , Metabolismo , Cardiomegalia , Metabolismo , Células Cultivadas , Cistationina gamma-Liasa , Metabolismo , Sulfuro de Hidrógeno , Metabolismo , MicroARNs , Metabolismo , Miocitos Cardíacos , Metabolismo , Cadenas Pesadas de Miosina , Metabolismo , Factores de Transcripción NFATC , Metabolismo , Péptido Natriurético Encefálico , Metabolismo , Proteínas del Tejido Nervioso , Metabolismo , Transducción de Señal

RESUMEN

tonomous Prefecture,Enshi Hubei 445000,China;2.Graduate School,Guangdong Medical College,Zhanjiang Guangdong 524001,China) Objective To study the relationship of miRNA-133a-3p and hypoxia in rat cardiomyocytes,and to detect the expression of the probable target genes.Methods SD rats cardiomyocytes were cultivated,and treated in hypoxia condition(37 ℃,5%CO2 ,1%O2 )for 0 hour,4 hours,8 hours,12 hours,16 hours,20 hours and 24 hours.The miRNA-133a-3p level was detected by qRT-PCR,and its biological information and target genes were predicted by targetScan picTar and miRanda,etc.Then we carried out qRT-PCR and Western blot to detect the expression of the probable target genes.Results miRNA-133a-3p expression was much higher in 4 hours hypoxia (P =0.000),then its expression gradually declined,and stabilized after 12 hours hypoxia.Bioinformatics prediction found that TGF-β1 may be its target.The mR-NA expression of TGF-β1 was obviously higher in 4 hours hypoxia than those in 0 hour and 24 hours(P Objective To study the relationship of miRNA-133a-3p and hypoxia in rat cardiomyocytes,and to detect the expression of the probable target genes.Methods SD rats cardiomyocytes were cultivated,and treated in hypoxia condition(37 ℃,5%CO2 ,1%O2 )for 0 hour,4 hours,8 hours,12 hours,16 hours,20 hours and 24 hours.The miRNA-133a-3p level was detected by qRT-PCR,and its biological information and target genes were predicted by targetScan picTar and miRanda,etc.Then we carried out qRT-PCR and Western blot to detect the expression of the probable target genes.Results miRNA-133a-3p expression was much higher in 4 hours hypoxia (P =0.000),then its expression gradually declined,and stabilized after 12 hours hypoxia.Bioinformatics prediction found that TGF-β1 may be its target.The mR-NA expression of TGF-β1 was obviously higher in 4 hours hypoxia than those in 0 hour and 24 hours(P <0.05).And the protein expression of TGF-β1 was significantly higher in 4 hours hypoxia(P <0.05).Conclusion miRNA-133a-3p may participate in cardiomyocytes necrosis in rats through regulating TGF-β1.

RESUMEN

AIM:To investigate the effects of ursolic acid ( UA) on the migration and invasion of human lung cancer cell line A549, and to explore its mechanism .METHODS:The cell viability was detected by MTT assay .The ex-pression of miRNA-133a was detected in the A549 cells treated with UA by real-time PCR.The miRNA-133a mimics and inhibitor were transfected into the A 549 cells, and the transfection efficiency was analyzed by real-time PCR.The cell mi-gratory and invasive abilities were determined by wound healing and Transwell methods , respectively .RESULTS:The via-bility of the human lung cancer A549 cells was significantly inhibited by UA in a dose-dependent manner (P<0.05).IC50 of UA (24 h) for lung cancer A549 cells was 31.04 μmol/L.UA treatment significantly inhibited the migratory and inva-sive abilities of A549 cells in a concentration-dependent manner , accompanied by significantly elevation of miRNA-133a expression.The mimics and inhibitor of miRNA-133a significantly upregulated and downregulated the expression of miRNA -133a in the transfected A549 cells, respectively.In addition, the viability of the A549 cells was decreased extremely after tansfected with the miRNA-133a mimics (P<0.01), so did the results of the cell migration and invasion test .The A549 cells tansfected with the miRNA-133a inhibitor showed an opposite changes of the cell viability , migration and invasion . CONCLUSION:UA inhibited the viability , migration and invasion of lung cancer A 549 cells by elevating the expression of miRNA-133a.