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1.
Journal of Integrative Medicine ; (12): 47-61, 2023.
Article in English | WPRIM | ID: wpr-971646

ABSTRACT

OBJECTIVE@#Huangqi Decoction (HQD), a classical traditional Chinese medicine formula, has been used as a valid treatment for alleviating liver fibrosis; however, the underlying molecular mechanism is still unknown. Although our previous studies showed that microRNA-663a (miR-663a) suppresses the proliferation and activation of hepatic stellate cells (HSCs) and the transforming growth factor-β/small mothers against decapentaplegic (TGF-β/Smad) pathway, whether long noncoding RNAs (lncRNAs) are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported. The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.@*METHODS@#The expression levels of lnc-C18orf26-1, miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction. HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs. The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs. Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs. RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs. Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA. The expression levels of collagen α-2(I) chain (COL1A2), α-smooth muscle actin (α-SMA) and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.@*RESULTS@#Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a. Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation, downregulated TGF-β1-stimulated α-SMA and COL1A2 expression, and inhibited the TGF-β1/Smad signaling pathway. HQD suppressed the proliferation and activation of HSCs. HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs. Further studies showed that HQD inhibited the expression of COL1A2, α-SMA, TGF-β1, TGF-β type I receptor (TGF-βRI) and phosphorylated Smad2 (p-Smad2) in HSCs, and these effects were reversed by miR-663a inhibitor treatment.@*CONCLUSION@#Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis. HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.


Subject(s)
Humans , Transforming Growth Factor beta/pharmacology , Transforming Growth Factor beta1/metabolism , RNA, Long Noncoding/pharmacology , Drugs, Chinese Herbal/pharmacology , MicroRNAs/genetics , Hepatic Stellate Cells/pathology , Liver Cirrhosis/metabolism , Cell Proliferation , Transforming Growth Factors/pharmacology
2.
Journal of Central South University(Medical Sciences) ; (12): 1252-1259, 2023.
Article in English | WPRIM | ID: wpr-1010349

ABSTRACT

As a crucial regulatory molecule in the context of vascular stenosis, transforming growth factor-β (TGF-β), plays a pivotal role in its initiation and progression. TGF-β, a member of the TGF-β superfamily, can bind to the TGF-β receptor and transduce extracellular to intracellular signals through canonical Smad dependent or noncanonical signaling pathways to regulate cell growth, proliferation, differentiation, and apoptosis. Restenosis remains one of the most challenging problems in cardiac, cerebral, and peripheral vascular disease worldwide. The mechanisms for occurrence and development of restenosis are diverse and complex. The TGF-β pathway exhibits diversity across various cell types. Hence, clarifying the specific roles of TGF-β within different cell types and its precise impact on vascular stenosis provides strategies for future research in the field of stenosis.


Subject(s)
Humans , Transforming Growth Factor beta/metabolism , Constriction, Pathologic , Signal Transduction , Cell Differentiation , Vascular Diseases , Transforming Growth Factors , Transforming Growth Factor beta1
3.
Journal of Zhejiang University. Medical sciences ; (6): 646-652, 2023.
Article in English | WPRIM | ID: wpr-1009930

ABSTRACT

Transforming growth factor (TGF)-β is a group of cytokines with anti-inflammatory effects in the TGF family, which participates in the development of stress and depression-related mechanisms, and plays roles in the regulation of inflammatory response in depression and the recovery of various cytokine imbalances. The core symptoms of depression is associated with TGF-β level, and the psychological symptoms of depression are related to TGF-β gene polymorphism. Various antidepressants may up-regulate TGF-β level through the complex interaction between neurotransmitters and inflammatory factors, inhibiting inflammatory response and regulating cytokine imbalance to improve depressive symptoms. Studies have shown that recombinant TGF-β1 protein has beneficial effects in mouse depression models, indicating TGF-β1 might be a potential therapeutic target for depression and nasal sprays having the advantage of being fast acting delivery method. This article reviews the research progress on dynamic changes of TGF-β level before and after depression treatment and the application of TGF-β level as an indicator for the improvement of depressive symptoms. We provide ideas for the development of new antidepressants and for the evaluation of the treatment efficacy in depression.


Subject(s)
Animals , Mice , Transforming Growth Factor beta/metabolism , Transforming Growth Factor beta1 , Depression , Cytokines , Antidepressive Agents/therapeutic use , Transforming Growth Factors
4.
São Paulo; s.n; s.n; 2022. 116 p. tab, graf.
Thesis in English | LILACS | ID: biblio-1378343

ABSTRACT

Stem cells are undifferentiated cells that can be distinguished from others by their ability to self-renew and to differentiate into new specific cell types. Mesenchymal stem cells (MSC) are adult stem cells that can be obtained from different sources, such as adipose tissue, bone marrow, dental pulp, and umbilical cord. They can either replicate, originating new identical cells, or differentiate into cells of mesodermal origin and from other germ layers. MSC have been studied as new tools for regenerative therapy. Although encouraging results have been demonstrated, MSC-based therapies still face a great barrier: the difficulty of isolating these cells from heterogeneous environments. MSC are currently characterized by immunolabelling through a set of multiple surface membrane markers, including CD29, CD73, CD90 and CD105, which are also expressed by other cell types. Hence, the present work aimed to identify new specific biomarkers for the characterization of human MSC using DNA aptamers produced by the SELEX (Systematic Evolution of Ligands by EXponential Enrichment) technique. Our results showed that MSC from different origins bound to DNA candidate aptamers, that is, DNA or RNA oligonucleotides selected from random libraries that bind specifically to biological targets. Aptamer-bound MSC could be isolated by fluorescenceactivated cell sorting (FACS) procedures, enhancing the induction of differentiation into specific phenotypes (chondrocytes, osteocytes and adipocytes) when compared to the whole MSC population. Flow cytometry analyses revealed that candidate aptamers bound to 50% of the MSC population from dental pulp and did not present significant binding rates to human fibroblasts or lymphocytes, both used as negative control. Moreover, immunofluorescence images and confocal analyses revealed staining of MSC by aptamers localized in the surfacemembrane of these cells. The results also showed internal staining of human monocytes by our investigated aptamers. A non-specific control aptamer (CNTR APT) obtained from the random pool was then utilized to compare the specificity of the aptamers bound to the analyzed non-apoptotic cells, showing no staining for MSC. However, 40% of the monocytes bound to the CNTR APT. Normalized data based on the cells bound to candidate aptamers compared to those bound to the CNTR APT, revealed a 10 to 16-fold higher binding rate for MSC against 2-fold for monocytes. Despite its low specificity, monocyte-aptamer binding occurs probably due to the expression of shared markers with MSC, since monocytes are derived from hematopoietic stem cells and are important for the immune system ability to internalize/phagocyte external molecules. Given that, we performed a pull-down assay followed by mass spectrometry analysis to detect which MSC-specific protein or other target epitope not coexpressed by monocytes or the CNTR APT would bind to the candidate aptamer. Distinguishing between MSC and monocyte epitopes is important, as both cells are involved in immunomodulatory effects after MSC transplantations. ADAM17 was found to be a target of the APT10, emerging as a possible biomarker of MSC, since its involvement in the inhibition of the TGF signaling cascade, which is responsible for the differentiation of MSC. Thus, MSC with a higher stemness profile should overexpress the protein ADAM17, which presents a catalytic site with affinity to APT10. Another target of Apt 10 is VAMP3, belonging to a transmembrane protein complex that is involved in endocytosis and exocytosis processes during immune and inflammatory responses. Overall, proteins identified as targets of APT10 may be cell surface MSC biomarkers, with importance for MSC-based cell and immune therapies


Células tronco são células indiferenciadas que podem ser distinguidas de outros tipos celulares por meio da habilidade de se auto renovarem e de se diferenciarem em novos tipos celulares. Células tronco mesenquimais (MSC) são células tronco adultas encontradas em diferentes tecidos como tecido adiposo, polpa de dente e cordão umbilical. Estas células podem se autodividir em células idênticas ou se diferenciarem em células de origem mesodermal. Estas células têm sido estudadas em novas aplicações que envolvem terapia regenerativas. Embora resultados encorajadores tenham sido demonstrados, terapias que utilizam MSC ainda encontram uma grande barreira: a dificuldade no isolamento destas células a partir de um ambiente heterogêneo. MSC são caracterizadas por populações positivas em ensaios de imunomarcação para os epítopos membranares CD29, CD73, CD90 e CD105, presentes também em outros tipos celulares. Assim, o presente trabalho tem o objetivo de identificar novos biomarcadores de MSC de origem humana, utilizando aptâmeros de DNA produzidos pela técnica SELEX (Systematic Evolution of Ligands by EXponential Enrichment) como ferramenta. Nossos resultados mostraram que MSC de diferentes origens ligam-se a aptâmeros (oligonucleotídeos de DNA ou RNA que atuam como ligantes específicos de alvos moleculares) de DNA candidatos que atuam no isolamento de MSC por meio da técnica FACS de separação celular, promovendo uma maior indução de diferenciação em células específicas (condrócitos, osteócitos e adipócitos) comparada com a população total de MSC. Análises de citometria de fluxo mostraram que os aptâmeros candidatos se ligam a 50% das MSC de polpa de dente e não apresentam taxa de ligação significante para fibroblastos e linfócitos de origem humana - utilizados como controles negativo. Além domais, imagens de imunofluorescência e confocal mostraram ligação na superfície da membrana de MSC e a marcação interna de monócitos a estes aptâmeros. Portanto, um aptâmero controle (CNTR APT) foi utilizado para comparar a especificidade dos aptâmeros ligados a células viáveis, mostrando a não ligação deste aptâmero a MSC. Porém, 40% da população de monócitos ligou-se ao CNTR APT. Uma normalização baseada na comparação entre as taxas de ligação entre células ligadas com aptâmeros candidatos e o aptâmero controle gerou uma taxa de especificidade entre 10-16 vezes maior para MSC contra 2,5 vezes para os monócitos. Deste modo, embora os resultados tenham mostrado uma taxa de ligação entre monócitos e aptâmeros, as MSC ligadas aos aptâmeros candidatos possuem uma maior taxa de especificidade devido a uma maior presença de antígenos que são expressos em ambas as células. Um ensaio de Pull Down seguido de espectrometria de massas foi utilizado para a identificação de biomarcadores que se ligariam aos aptâmeros candidatos, e que não seriam co-expressos por monócitos e por antígenos ligados ao aptâmero controle. Deste modo, a proteína ADAM17 foi identificada nas amostras de APT10 ligadas às MSC. Tal proteína está relacionada à inibição de uma cascata de sinalização da família de proteínas TGF, responsável pela diferenciação de MSC. Assim, MSC com maior potencial tronco deveriam expressar ADAM17 em maior quantidade. Tal proteína apresenta um sítio catalítico que demonstra interagir com o APT10, de acordo com predição Docking entre proteína e DNA. Foi identificada também, a proteína VAMP3, que pertence a um complexo proteico transmembranar responsável pelos processos de endocitose e exocitose, e que podem ter um papel importante na liberação de citocinas e outras moléculas relacionadas às respostas imune e inflamatórias. Deste modo, o APT10 identificou proteínas importantes que devem estar relacionas com a melhora de imunoterapias que utilizam MSC


Subject(s)
Stem Cells , Biomarkers/analysis , SELEX Aptamer Technique/instrumentation , Mesenchymal Stem Cells/classification , ADAM17 Protein/pharmacology , Patient Isolation , Mass Spectrometry/methods , Staining and Labeling/methods , Transplantation/adverse effects , Umbilical Cord , DNA/agonists , Transforming Growth Factors/agonists , Cell Separation/instrumentation , Cytokines/adverse effects , Adipocytes/metabolism , Chondrocytes/classification , Scientists for Health and Research for Development , Adult Stem Cells/classification , Fibroblasts/chemistry , Flow Cytometry/instrumentation , Germ Layers , Antigens/adverse effects
5.
São Paulo; s.n; s.n; 2022. 62 p. graf.
Thesis in Portuguese | LILACS | ID: biblio-1378655

ABSTRACT

As Células-Tronco Mesenquimais (CTMs), são células multipotentes, presentes em diversos tecidos, sendo bastante estudada devido sua capacidade imunorregulatória por meio da liberação de fatores solúveis. Fatores estes que atuam sobre as funções de células do sistema imunitário. Simultaneamente, estudos indicam que os compostos flavonoides, em destaque a Delfinidina, presente em alguns frutos e flores, possuem atuação anti-inflamatória e inibitória sobre células do sistema imunitário. Todavia, são escassos os estudos em relação entre a capacidade imunorregulatória da CTM e a influência da Delfinidina, sendo este o objetivo deste estudo. Inicialmente, a Delfinidina 3-O-ß-D-glicosídeo foi escolhido, devido a sua maior estabilidade e a dose de 50 µM foi selecionada após análise por citometria de fluxo que mostrou aumento da fase proliferativa do ciclo celular. Posteriormente ao realizar análise da produção de fatores solúveis pelas CTM, os resultados mostraram aumento da produção de IL-10, TGF-ß e Oxido nítrico pelas CTM tratadas com Delfinidina. Bem como, diminuição da expressão de p-NF-κB/NF-κB pelas CTMs tratadas com Delfinidina, quando avaliadas por Wersten Blot. Adicionalmente, para analisar a Delfinidina sobre os efeitos imunorregulatórios da CTM sob macrófagos (RAW 264.7), célula esta, importante no sistema imune inato. Foram realizadas culturas condicionadas, com posterior análise da produção de fatores solúveis, os resultados mostraram aumento da produção de IL-10, e diminuição da produção de TNF-α, IL-1α e IL-12 pelos macrófagos, nas culturas condicionadas. Assim como, diminuição da expressão do fator p-NF-κB/NF-κB pelos macrófagos nas culturas condicionadas, quando avaliadas por Wersten Blot. Ademais, ao analisar a atividade metabólica dos macrófagos por ensaio de MTT, os resultados mostraram que as culturas condicionadas e a Delfinidina per si foi capaz de diminuir a atividade metabólica, sem alterar os efeitos anti-inflamatórios sobre a célula. Em síntese, a Delfinidina mostrou acentuar a atuação imunorregulatória da CTM sobre a linhagem macrofágica, célula esta, de grande importância para o sistema imune inato


Mesenchymal Stem Cells (MSCs) are multipotent cells present in various tissues, being widely studied due to their immunoregulatory capacity through the release of soluble factors. These factors act on the functions of cells of the immune system. Simultaneously, studies indicate that flavonoid compounds, especially Delphinidin, present in some fruits and flowers, have anti inflammatory and inhibitory effects on immune system cells. However, there are few studies on the relationship between the immunoregulatory capacity of MSC and the influence of Delphinidin, which is the objective of this study. Initially, Delphinidin 3-O-ß-D-glycoside was chosen due to its greater stability and the 50 µM dose was selected after analysis by flow cytometry which showed an increase in the proliferative phase of the cell cycle. Subsequently, when analyzing the production of soluble factors by MSCs, the results showed an increase in the production of IL-10, TGF-ß and nitric oxide by MSCs treated with Delphinidin. As well as decreased expression of p-NF-κB/NF-κB by MSCs treated with Delphinidin, when evaluated by Wersten Blot. Additionally, to analyze Delphinidin on the immunoregulatory effects of MSC on macrophages (RAW 264.7), this cell is important in the innate immune system. Conditioned cultures were performed, with subsequent analysis of the production of soluble factors, the results showed an increase in the production of IL-10, and a decrease in the production of TNF-α, IL-1α and IL-12 by macrophages, in the conditioned cultures. As well as decreased expression of p-NF-κB/NF-κB factor by macrophages in conditioned cultures, when evaluated by Wersten Blot. Furthermore, when analyzing the metabolic activity of macrophages by MTT assay, the results showed that conditioned cultures and Delphinidin itself was able to decrease the metabolic activity, without altering the anti-inflammatory effects on the cell. In summary, Delphinidin has shown to enhance the immunoregulatory action of MSC on the macrophage lineage, a cell that is of great importance for the innate immune system


Subject(s)
Flavonoids/analysis , Immune System , Transforming Growth Factors , Interleukin-1/adverse effects , Interleukin-10/adverse effects , Mesenchymal Stem Cells/classification , Flow Cytometry/instrumentation , Anti-Inflammatory Agents/administration & dosage
6.
Int. j. morphol ; 39(1): 18-24, feb. 2021. ilus, tab
Article in English | LILACS | ID: biblio-1385293

ABSTRACT

SUMMARY: Diabetes is a metabolic disorder characterized by high blood sugar levels and it causes complications in many systems, including the reproductive system. As a result of diabetic conditions, one of the mechanisms that can cause repression of reproductive activity is testicular oxidant stress. The identification of diabetes on the cell signaling molecules axis is still under discussion. The aim of this study was to determine the effect of Transforming Growth Factor (TGFβ), Nuclear Factor kappa B (NF-kB), Heat-schock 90β (HSP90β) signal pathways and E-cadherin cell adhesion molecule on infertility in diabetic rat testicular tissue. In our study, includes histological, molecular and biochemical analysis of testicular tissue removed at the end of the 2 weeks experiment period. A total of 14 adult male rats were divided as control and diabetes. No intervention was given to 7 male rats in the control group. For the diabetic group, 7 male rats were injected by intraperitoneal with a single dose of 55 mg/kg streptozotocin (STZ). TGFβ, NF-kB, HSP90β and E-cadherin proteins were immunohistochemically studied to investigate possible tissue damage, inflammatory process, cell stabilization and integrity due to diabetes. In order to determine oxidant stress, lipid peroxidation product malondialdehyde (MDA), glutathione (GSH) and glutathione peroxidase (GPx) analyzes were performed. Fibrosis, inflammatory changes and loss of spermatogenetic series are prominent findings in the diabetic group. On analysis of all the samples with immunostaining, in the diabetic group, TGFβ and NF-kB immunoexpression significantly increased, while Hsp90β and E-cadherin immunoexpression significantly decreased compared with control groups. Experimental diabetes was found to cause fibrosis, inflammation, disrupting cell adhesion and stabilization in testicular tissue. These results suggest that cellular therapy studies are needed for possible damage.


RESUMEN: La diabetes es una enfermedad metabólica caracterizada por niveles altos de azúcar en sangre y causa complicaciones en muchos sistemas, incluido el sistema reproductivo. Como resultado de las condiciones diabéticas, uno de los mecanismos que puede causar alteraciones en la actividad reproductiva es el estrés oxidativo testicular. La identificación de la diabetes en el eje de las moléculas de señalización celular aún está en discusión. El objetivo de este estudio fue determinar el efecto del factor de crecimiento transformante (TGFβ), el factor nuclear kappa B (NF-kB), las vías de señalización de Heat-Schock 90b (HSP90β) y la molécula de adhesión celular de E-cadherina sobre la infertilidad en testículo de rata diabética. Al término de dos semanas se realizaron análisis histológico, molecular y bioquímico del tejido testicular extraído. Las 7 ratas macho del grupo control no fueron intervenidas. Para el grupo de diabéticos, 7 ratas macho fueron inyectadas por vía intraperitoneal con una dosis única de 55 mg / kg de estreptozotocina (STZ). Se estudiaron inmunohistoquímicamente las proteínas TGFβ, NF-kB, HSP90β y E-cadherina para investigar el posible daño tisular, el proceso inflamatorio, la estabilización celular y la integridad debido a la diabetes. Para determinar el estrés oxidativo, se realizaron análisis del producto de peroxidación lipídica malondialdehído (MDA), glutatión (GSH) y glutatión peroxidasa (GPx). La fibrosis, los cambios inflamatorios y la pérdida de series espermatogenéticas son hallazgos destacados en el grupo de ratas diabéticas. En el análisis de todas las muestras con inmunotinción, en el grupo diabético, la inmunoexpresión de TGFβ y NF-kB aumentó significativamente, mientras que la inmunoexpresión de Hsp90β y e-cadherina disminuyó significativamente en comparación con los grupos control. Se encontró que la diabetes experimental causa fibrosis, inflamación, alteración de la adhesión celular y estabilización en el tejido testicular. Estos resultados sugieren que son necesarios estudios de terapia celular para verificar posibles daños.


Subject(s)
Animals , Male , Rats , Testis/pathology , Diabetes Mellitus, Experimental/metabolism , Testis/metabolism , Immunohistochemistry , Transforming Growth Factors/metabolism , Cadherins/metabolism , NF-kappa B/metabolism , HSP90 Heat-Shock Proteins/metabolism
7.
Article in English | LILACS, BBO | ID: biblio-1180856

ABSTRACT

ABSTRACT Objective: To compare Transforming growth factor-β1 (TGF-β1) expression in various L-PRF concentrations on the hDPSC differentiation process. Material and Methods: hDPSC cell cultures were subjected to serum starvation by reducing FBS levels in the hDPSC culture media. Lysate PRF was obtained from the PRF gel, which was then incubated at 4°C for 24 h. The supernatant was dried, transferred to a 2-ml Eppendorf tube, and stored at −20°C. The evaluation of TGF-β1 expression in 1%, 5%, 10%, and 25% L-PRF samples and 10% FBS (control) during the process of hDPSC differentiation was quantified using an ELISA reader on day 7. The expression of TGF-β1 was subjected to a one-way ANOVA test, followed by Bonferroni's post hoc test with significant values (p<0.05). Results: Significant differences were noted in TGF-β1 expression between 1%, 5%, 10%, and 25% L-PRF and the control group (10% FBS). The highest TGF-β1 expression occurred with 25% L-PRF (0.645 ± 0.048), followed by 10% L-PRF (0.461 ± 0.035), 10% FBS (0.374 ± 0.013), 5% L-PRF (0.275 ± 0.045), and the lowest expression was with 1% L-PRF (0.160 ± 0.045). Conclusion: The best result of TGF-B1 expression in hDPSC differentiation was in the 25% L-PRF group.


Subject(s)
Humans , Cell Culture Techniques , Culture Media/analysis , Dental Pulp , Platelet-Rich Fibrin/microbiology , Enzyme-Linked Immunosorbent Assay , Transforming Growth Factors , Cell Differentiation/immunology , Analysis of Variance , Indonesia
8.
Journal of Nutrition and Health ; : 1-12, 2020.
Article in Korean | WPRIM | ID: wpr-811256

ABSTRACT

PURPOSE: Diabetic nephropathy is one of the most important diabetic complications prompted by chronic hyperglycemia, characterized by glomerulosclerosis, tubular fibrosis, and it eventually causes kidney failure. Nobiletin is a polymethoxyflavone present in tangerine and other citrus peels, and has anti-cancer and anti-inflammatory effects. This study investigated the effects of nobiletin on glomerular fibrosis through inhibition of the transforming growth factor (TGF)-β1-Src-caveolin-1 pathway.METHODS: Human renal mesangial cells (HRMC) were incubated in media containing 33 mM glucose with or without 1–20 uM nobiletin for 3 day. The cellular expression levels of fibrogenic collagen IV, fibronectin, connective tissue growth factor (CTGF), TGF-β1, Src and caveolin-1 were all examined. In addition, TGF-β1, Src and caveolin-1 proteins were screened to reveal the relationship among TGF-β1-Src-caveolin-1 signaling in glomerular fibrosis.RESULTS: High glucose promoted the production of collagen IV, fibronectin and CTGF in HRMC, which was inhibited in a dose dependent manner by 1–20 uM nobiletin. The Western blot data showed that high glucose elevated the expression of TGF-β1, Src, caveolin-1 and Rho GTPase. When nobiletin was treated to the HRMC exposed to high glucose, the expression of TGF-β1-Src-caveolin-1 was dampened. Finally, TGF-β1-Src-caveolin-1 signaling pathway was activated in high glucose-exposed HRMC, and such activation was encumbered by nobiletin.CONCLUSION: These result demonstrated that nobiletin blunted high glucose-induced extracellular matrix accumulation via inhibition of the TGF-β1-Src-caveolin-1 related intracellular signaling pathway. Nobiletin may be a potent renoprotective agent to counteract diabetes-associated glomerular fibrosis that leads to kidney failure.


Subject(s)
Humans , Blotting, Western , Caveolin 1 , Citrus , Collagen , Connective Tissue Growth Factor , Diabetes Complications , Diabetic Nephropathies , Extracellular Matrix , Fibronectins , Fibrosis , Glucose , GTP Phosphohydrolases , Hyperglycemia , Mesangial Cells , Renal Insufficiency , Transforming Growth Factors
9.
Araçatuba; s.n; 2019. 76 p. ilus, tab, graf.
Thesis in Portuguese | LILACS, BBO | ID: biblio-1399507

ABSTRACT

Na presença de Diabetes Mellitus, a doença periodontal (DP) pode acentuar o aparecimento de complicações e aumentar sua prevalência. O objetivo desta pesquisa foi avaliar se o Fator de Crescimento Transformador -ß do tipo 1 (TGFß1, sigla em inglês) e Metaloproteinase de matriz -9 (MMP-9, sigla em ingês) estão modulando a formação de componentes de matriz extracelular em fibroblastos gengivais (FG) de camundongos diabéticos e com DP. Os 24 animais foram separados em 4 grupos, e então foram tratados com estreptozotocina para indução de diabetes e foi realizado indução da DP através de ligadura com fio de seda. Após a eutanásia e coleta da gengiva, FG dos camundongos foram cultivados. A expressão gênica de RNAm e produção de proteína para MMP-9, TGF-ß1 e colágeno tipo 1 (Col1a1) foram avaliadas. Eles também foram tratados com inibidores de MMP-9 e TGF-ß1, e dexametasona (Dexa) por 24h. FG de animais normais e diabéticos com DP induzida tiveram expressão aumentada de MMP-9 e TGF-ß1 em 6 e 24h, diminuindo em 48h. Expressão gênica de Col1a1 foi diminuída em FG com DP em 6, 24 e 48h. Inibidor de MMP-9 (MMP-9i) bloqueou a expressão gênica de MMP-9 e de TGF-ß1 em FG normais e diabéticos em 24h. A expressão de col1a1 foi inibida pelo MMP-9i somente em FG normais, mas não em diabéticos. A expressão de receptor de TGF-ß do tipo 1 (TGF-ß R1) foi aumentada em FG normais e diabéticos, porém MMP-9i diminuiu esta expressão somente em FG diabéticos. Inibidor de TGF-ß (SB431542) e Dexa interromperam a expressão gênica de MMP-9, TGF-ß1 e col1a1 em FG normais e diabéticos com DP em 24h. TGF-ß R1 também foi inibido em FG normais e diabéticos com DP em 24h de tratamento com SB431542. Juntos, esses achados sugerem que indivíduos diabéticos têm síntese aumentada de TGF-ß e MMP-9 em FG durante a DP devido a ativação do TGF-ß R1 via ALK5 e presença de MMP-9. Por outro lado, o aumento na expressão de colágeno é devido a ativação de TGF-ß R1 em FG normais e diabéticos, e presença de MMP-9 em FG normais, somente(AU)


In presence of Diabetes Mellitus, periodontal disease (PD) can accentuate complication appearance and increase prevalence. The aim of this study was to evaluate whether Transforming Growth factor-ß 1 (TGF-ß1) and Matrix Metaloproteinase-9 (MMP-9) are modulating the extracellular matrix components in gingival fibroblasts (GF) from diabetic mice (D) with PD. 24 animals were divided in 4 groups then they were treated with streptozotocin to diabetes induction and DP was inducted through a ligature with silk thread. After euthanasia and gingiva harvest, mice GF were cultured. MMP-9, TGF- ß1, and type 1 Collagen (col1a1) mRNA expression and protein production were evaluated. They were also treated with MMP-9 and TGF-ß inhibitors, and dexamethasone (Dexa) for 24h. Normal and diabetic GF from PD-induced mice had increased MMP-9 and TGF-ß1 expression at 6 and 24 h decreasing at 48 h. Col1a1 gene expression was decreased in normal GF with PD at 6, 24 and 48 h. In diabetic GF col1a1 expression was inhibited at 6, 24 and 48 h. MMP-9 inhibitor (MMP-9i) blocked the MMP-9 expression, and TGF-ß1 gene expression in normal and diabetic GF at 24 h. The col1a1 gene expression was inhibited by MMP-9i only in normal GF but not in diabetic. TGF-ß R1 was increased in normal and diabetic GF with PD, but MMP9i decreased this expression only in diabetic GF. TGF-ß inhibitor (SB431542) and Dexa abrogated MMP-9, TGF-ß1 and col1a1 expression in normal and diabetic GF with PD at 24 h. TGF-ß R1 was also inhibited in normal and diabetic GF with PD after 24 h of treatment with SB431542. Taken together, these findings suggests that diabetic individuals has increased synthesis of TGF-ß and MMP-9 in GF during PD due to TGF-ß R1 activation via Alk5 and presence of MMP-9. On the other hand, increased in the col1a1 expression is due to TGF-ß R1 activation in normal and diabetic GF and presence of MMP-9 in normal GF(AU)


Subject(s)
Animals , Mice , Periodontal Diseases , Transforming Growth Factors , Matrix Metalloproteinases , Diabetes Mellitus , Fibroblasts , Gingiva , Dexamethasone , Collagen , Collagen Type I , Diabetes Mellitus/epidemiology , Matrix Metalloproteinase Inhibitors
10.
Allergy, Asthma & Respiratory Disease ; : 150-157, 2019.
Article in Korean | WPRIM | ID: wpr-762190

ABSTRACT

PURPOSE: Atopic dermatitis (AD) is the most common chronic and relapsing inflammatory skin disease with skin barrier defects and altered immune responses. Chronic inflammation leads to irreversible fibrosis in the skin and there is no treatment to completely abolish the inflammation and fibrosis. To prevent or treat the chronic process of AD, it is necessary to develop a murine model of AD that reflects the chronic process to identify the mechanism. The aims of this study were to develop a chronic AD model with a crude extract Aspergillus fumigatus (Af) antigen. METHODS: We applied Af extract (40 µg) epicutaneously to the dorsal skin of BALB/c mice for 5 consecutive days per week during a period of 5 weeks for a chronic AD model, and 5 consecutive days repeatedly with 2 weeks interval for an acute AD model. RESULTS: The clinical score and transepidermal water loss were more increased in the chronic AD model than in the acute AD model. Histologic findings showed that more increased epidermal thickness, neutrophil infiltration and hyperkeratosis in the chronic model than in the acute model. Skin fibrosis was more prominent in the chronic model than in the acute model. The mRNA expression levels of transforming growth factor (TGF)-β, thymic stromal lymphopoietin, and interleukin-33 were increased in the skin of the chronic model compared to the acute model. The levels of total IgE, Af-specific IgE, IgG1, and IgG2a were significantly increased in the chronic model compared to controls. CONCLUSION: The Af-induced chronic AD model showed prominent fibrosis and increased TGF-β expression in the skin, which suggests that these models may be useful in the research for the mechanism of the chronic process in AD.


Subject(s)
Animals , Mice , Aspergillus fumigatus , Aspergillus , Dermatitis, Atopic , Fibrosis , Immunoglobulin E , Immunoglobulin G , Inflammation , Interleukin-33 , Neutrophil Infiltration , RNA, Messenger , Skin , Skin Diseases , Transforming Growth Factors , Water
11.
Yonsei Medical Journal ; : 561-569, 2019.
Article in English | WPRIM | ID: wpr-762078

ABSTRACT

PURPOSE: Liver fibrosis is a major cause of morbidity and mortality and the outcome of various chronic liver diseases. Activation of hepatic stellate cells (HSCs) is the key event in liver fibrosis. Studies have confirmed that miR-140-3p plays a potential regulatory effect on HSC activation. However, whether miR-140-3p mediates the liver fibrosis remains unknown. MATERIALS AND METHODS: Expression of miR-140-3p was detected by real-time quantitative PCR (qPCR). Cell proliferation was measured by MTT, while cell apoptosis rate was determined via flow cytometry. Western blot assay was used to detect the expression of cleaved PARP. The fibrogenic effect was evaluated by expression of α-smooth muscle actin and desmin. Functional experiments were performed in transforming growth factor β1 (TGF-β1)-induced HSC-T6 cells with transfection of anti-miR-140-3p and/or siPTEN. Target binding between miR-140-3p and PTEN was predicted by the TargetScan database and identified using luciferase reporter assay and RNA immunoprecipitation. RESULTS: TGF-β1 induced the activation of HSC-T6 cells, and miR-140-3p expression varied according to HSC-T6 cell activation status. Knockdown of miR-140-3p reduced cell proliferation and the expressions of α-SMA and desmin, as well as increased apoptosis, in TGF-β1-induced HSC-T6 cells, which could be blocked by PTEN silencing. Additionally, inactivation of the AKT/mTOR signaling pathway stimulated by miR-140-3p knockdown was abolished when silencing PTEN expression. PTEN was negatively regulated by miR-140-3p via direct binding in HSC-T6 cells. CONCLUSION: miR-140-3p is an important mediator in HSC-T6 cell activation, and miR-140-3p knockdown suppresses cell proliferation and fibrogenesis in TGF-β1-induced HSC-T6 cells, indicating that miR-140-3p may be a potential novel molecular target for liver fibrosis.


Subject(s)
Actins , Apoptosis , Blotting, Western , Cell Proliferation , Desmin , Flow Cytometry , Hepatic Stellate Cells , Immunoprecipitation , Liver Cirrhosis , Liver Diseases , Luciferases , Mortality , Polymerase Chain Reaction , RNA , Transfection , Transforming Growth Factors
12.
Journal of Korean Neurosurgical Society ; : 153-165, 2019.
Article in English | WPRIM | ID: wpr-788767

ABSTRACT

OBJECTIVE: Spinal cord injury (SCI) is a very serious health problem, usually caused by a trauma and accompanied by elevated levels of inflammation indicators. Stem cell-based therapy is promising some valuable strategies for its functional recovery. Nestin-positive progenitor and/or stem cells (SC) isolated from pancreatic islets (PI) show mesenchymal stem cell (MSC) characteristics. For this reason, we aimed to analyze the effects of rat pancreatic islet derived stem cell (rPI-SC) delivery on functional recovery, as well as the levels of inflammation factors following SCI.METHODS: rPI-SCs were isolated, cultured and their MSC characteristics were determined through flow cytometry and immunofluorescence analysis. The experimental rat population was divided into three groups : 1) laminectomy & trauma, 2) laminectomy & trauma & phosphate-buffered saline (PBS), and 3) laminectomy+trauma+SCs. Green fluorescent protein (GFP) labelled rPI-SCs were transplanted into the injured rat spinal cord. Their motilities were evaluated with Basso, Beattie and Bresnahan (BBB) Score. After 4-weeks, spinal cord sections were analyzed for GFP labeled SCs and stained for vimentin, S100β, brain derived neurotrophic factor (BDNF), 2’,3’-cyclic-nucleotide 3'-phosphodiesterase (CNPase), vascular endothelial growth factor (VEGF) and proinflammatory (interleukin [IL]-6, transforming growth factor [TGF]-β, macrophage inflammatory protein [MIP]-2, myeloperoxidase [MPO]) and anti-inflammatory (IL-1 receptor antagonis) factors.RESULTS: rPI-SCs were revealed to display MSC characteristics and express neural and glial cell markers including BDNF, glial fibrillary acidic protein (GFAP), fibronectin, microtubule associated protein-2a,b (MAP2a,b), β3-tubulin and nestin as well as antiinflammatory prostaglandin E2 receptor, EP3. The BBB scores showed significant motor recovery in group 3. GFP-labelled cells were localized on the injury site. In addition, decreased proinflammatory factor levels and increased intensity of anti-inflammatory factors were determined.CONCLUSION: Transplantation of PI-SCs might be an effective strategy to improve functional recovery following spinal cord trauma.


Subject(s)
Animals , Rats , Brain-Derived Neurotrophic Factor , Dinoprostone , Fibronectins , Flow Cytometry , Fluorescent Antibody Technique , Glial Fibrillary Acidic Protein , Inflammation , Islets of Langerhans , Laminectomy , Macrophages , Mesenchymal Stem Cells , Microtubules , Nestin , Neuroglia , Peroxidase , Regeneration , Spinal Cord Injuries , Spinal Cord , Stem Cell Transplantation , Stem Cells , Transforming Growth Factors , Vascular Endothelial Growth Factor A , Vimentin , Wounds and Injuries
13.
Intestinal Research ; : 527-536, 2019.
Article in English | WPRIM | ID: wpr-785863

ABSTRACT

BACKGROUND/AIMS: Transforming growth factor-β1 (TGF-β1) induction of epithelial-mesenchymal transition (EMT) is one of the mechanisms by which colorectal cancer (CRC) cells acquire migratory and invasive capacities, and subsequently metastasize. Parthenolide (PT) expresses multiple anti-cancer and anti-inflammatory activities that inhibit nuclear factor κB by targeting the IκB kinase complex. In the present study, we aimed to investigate whether PT can inhibit TGF-β1-induced EMT in CRC cell lines.METHODS: HT-29 and SW480 cell lines were used in the experiment. Cell viability was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and sub-G1 analysis was measured by flow cytometry. The induction of EMT by TGF-β1 and inhibition of the process by PT was analyzed by phase contrast microscopy, wounding healing, cellular migration and invasion assays, and Western blotting.RESULTS: TGF-β1 inhibits HT-29 cell proliferation, but has no effect on SW480 cell proliferation; different concentrations of TGF-β1 did not induce apoptosis in HT-29 and SW480 cells. PT attenuates TGF-β1-induced elongated, fibroblast-like shape changing in cells. PT inhibits TGF-β1-induced cell migration and cell invasion. In addition, other EMT markers such as β-catenin, Vimentin, Snail, and Slug were suppressed by PT, while E-cadherin was increased by PT.CONCLUSIONS: Our findings show that PT inhibits TGF-β1-induced EMT by suppressing the expression of the mesenchymal protein and increasing expression of the epithelial protein. These findings suggest a novel approach for CRC treatment by suppression of TGF-β1-induced EMT.


Subject(s)
Humans , Apoptosis , Blotting, Western , Cadherins , Cell Line , Cell Movement , Cell Proliferation , Cell Survival , Colorectal Neoplasms , Epithelial-Mesenchymal Transition , Flow Cytometry , Gastropoda , HT29 Cells , Microscopy, Phase-Contrast , Phosphotransferases , Snails , Transforming Growth Factors , Vimentin , Wounds and Injuries
14.
International Journal of Stem Cells ; : 400-409, 2019.
Article in English | WPRIM | ID: wpr-785834

ABSTRACT

BACKGROUND AND OBJECTIVES: The release of microvesicles (MVs) from mesenchymal stem cells (MSCs) has been implicated in intercellular communication, and may contribute to beneficial paracrine effects of stem cell-based therapies. We investigated the effect of administration of MSC-MVs on the therapeutic potential of carbon tetrachloride (CCL₄) induced liver fibrosis in rats.METHODS: Our work included: isolation and further identification of bone marrow MSC-MVs by transmission electron microscopy (TEM). Liver fibrosis was induced in rats by CCl4 followed by injection of prepared MSC-MVs in injured rats. The effects of MSC-MVs were evaluated by biochemical analysis of liver functions, RNA gene expression quantitation for collagen-1α, transforming growth factor β (TGF-β), interleukin-1β (IL-1β), vascular endothelial growth factor (VEGF) by real time reverse transcription PCR (RT-PCR) techniques. Finally histopathological examination of the liver tissues was assessed for all studied groups.RESULTS: BM-MSC-MVs treated group showed significant increase in serum albumin levels, VEGF quantitative gene expression (p < 0.05), while it showed a significant decrease in serum alanine transaminase (ALT) enzyme levels, quantitative gene expression of TGF-β, collagen-1α, IL-1β compared to CCL₄ fibrotic group (p < 0.05). Additionally, the histopathological assessment of the liver tissues of BM-MSC-MVs treated group showed marked decrease in the collagen deposition & improvement of histopathological picture in comparison with CCL₄ fibrotic group.CONCLUSIONS: Our study demonstrates that BM-MSC-MVs possess anti-fibrotic, anti-inflammatory, and pro-angiogenic properties which can promote the resolution of CCL₄ induced liver fibrosis in rats.


Subject(s)
Animals , Rats , Alanine Transaminase , Bone Marrow , Carbon Tetrachloride , Collagen , Gene Expression , Liver Cirrhosis , Liver , Mesenchymal Stem Cells , Microscopy, Electron, Transmission , Polymerase Chain Reaction , Reverse Transcription , RNA , Serum Albumin , Transforming Growth Factors , Vascular Endothelial Growth Factor A
15.
International Journal of Stem Cells ; : 170-182, 2019.
Article in English | WPRIM | ID: wpr-764050

ABSTRACT

OBJECTIVE: The aim of our study was to investigate the effect of Transforming growth factor beta-1 (TGF-β1) gene therapy on the surface markers, multilineage differentiation, viability, apoptosis, cell cycle, DNA damage and senescence of human Dental Pulp-derived Mesenchymal Stromal Cells (hDPSC). METHODS: hDPSCs were isolated from human teeth, and were cultured with 20% Fetal Bovine Serum (FBS) in minimum essential media-alpha (α-MEM). TGF-β1 gene transfer into hDPSCs was performed by electroporation method after the plasmid was prepared. The transfection efficiency was achieved by using western blot and flow cytometry analyses and GFP transfection. Mesenchymal stem cell (MSC) markers, multilineage differentiation, cell proliferation, apoptosis, cell cycle, DNA damage and cellular senescence assays were performed by comparing the transfected and non-transfected cells. Statistical analyses were performed using GraphPad Prism. RESULTS: Strong expression of TGF-β1 in pCMV-TGF-β1-transfected hDPSCs was detected in flow cytometry analysis. TGF-β1 transfection efficiency was measured as 95%. Western blot analysis showed that TGF-β1 protein levels increased at third and sixth days in pCMV-TGF-β1-transfected hDPSCs. The continuous TGF-β1 overexpression in hDPSCs did not influence the immunophenotype and surface marker expression of MSCs. Our results showed that TGF-β1 increased osteogenic and chondrogenic differentiation, but decreased adipogenic differentiation. Overexpression of TGF-β1 increased the proliferation rate and decreased total apoptosis in hDPSCs (p<0.05). The number of cells at “S” phase was higher with TGF-β1 transfection (p<0.05). Cellular senescence decreased in TGF-β1 transfected group (p<0.05). CONCLUSIONS: These results reflect that TGF-β1 has major impact on MSC differentiation. TGF-β1 transfection has positive effect on proliferation, cell cycle, and prevents cellular senescence and apoptosis.


Subject(s)
Humans , Aging , Apoptosis , Blotting, Western , Cellular Senescence , Cell Cycle , Cell Differentiation , Cell Proliferation , DNA Damage , Electroporation , Flow Cytometry , Genetic Therapy , Mesenchymal Stem Cells , Methods , Plasmids , Population Characteristics , Tooth , Transfection , Transforming Growth Factors
16.
Gut and Liver ; : 430-439, 2019.
Article in English | WPRIM | ID: wpr-763856

ABSTRACT

BACKGROUND/AIMS: The current study aims to investigate the protective effects of Bifidobacterium infantis on the abnormal immune response to inflammatory bowel disease (IBD) in dextran sodium sulfate (DSS)-induced colitis. METHODS: Eight-week-old BALB/c mice were separated into five groups at random (control, DSS, DSS+B9 [B. infantis 1×10⁹ CFU], DSS+B8 [B. infantis 1×10⁸ CFU], and DSS+B7 [B. infantis 1×10⁷ CFU]). Colitis was induced by 5% DSS ad libitum for 7 days, at which time we assessed weight, the disease activity index (DAI) score, and the histological damage score. The nuclear transcription factor Foxp3 (a marker of Treg cells), cytokines interleukin-10 (IL-10) and transforming growth factor β1 (TGF-β1), and related proteins (programmed cell death ligand 1 [PD-L1] and programmed cell death 1 [PD-1]) were detected by an immunohistochemical method and Western blot. RESULTS: B. infantis increased weight, decreased DAI scores and histological damage scores, increased the protein expression of Foxp3 (p<0.05) and cytokines IL-10 and TGF-β1 in mouse colon tissue (p<0.05), and increased the expression of PD-L1 in the treatment groups relative to that in the DSS group (p<0.05). The effect of B. infantis on Foxp3 and PD-L1 was dose dependent in the treatment groups (p<0.05). PD-L1 was positively correlated with Foxp3, IL-10, and TGF-β1. CONCLUSIONS: In a mouse model of IBD, B. infantis can alleviate intestinal epithelial injury and maintain intestinal immune tolerance and thus may have potential therapeutic value for the treatment of immune damage in IBD.


Subject(s)
Animals , Mice , Bifidobacterium , Blotting, Western , Cell Death , Colitis , Colon , Cytokines , Dextrans , Immune Tolerance , Inflammatory Bowel Diseases , Interleukin-10 , Methods , Models, Theoretical , Sodium , T-Lymphocytes, Regulatory , Transcription Factors , Transforming Growth Factors
17.
Clinical Endoscopy ; : 464-471, 2019.
Article in English | WPRIM | ID: wpr-763473

ABSTRACT

BACKGROUND/AIMS: Epithelial-mesenchymal transition (EMT) is a developmental process, wherein the epithelial cells show reduced intercellular adhesions and acquire migratory fibroblastic properties. EMT is associated with downregulation in epithelial marker expression, abnormal translocation of E-cadherin, and upregulation in mesenchymal marker expression. Here, we investigated the immunohistochemical (IHC) expression of EMT markers in early gastric cancer (EGC) between cancer and noncancer tissues. METHODS: Tissue samples were prospectively obtained from 19 patients with EGC that underwent endoscopic submucosal dissection (ESD). We compared the expression level of transforming growth factor (TGF)-β, vascular endothelial growth factor (VEGF), E-cadherin, α-smooth muscle actin (α-SMA), and vimentin between cancer and noncancer tissues using IHC. Among the 19 patients, 15 patients had follow-up biopsy at 3 months after ESD for EGC. RESULTS: Cancer tissues presented higher values of EMT mesenchymal markers (α-SMA/vimentin/TGF-β/VEGF) than the noncancerous tissues (p<0.05) that were significantly low after ESD (p<0.05). No significant correlation was reported for tumor location and initial Helicobacter pylori infection. CONCLUSIONS: The mesenchymal expression of EMT markers was higher in the cancerous tissues than in the noncancer tissues.


Subject(s)
Humans , Actins , Biopsy , Cadherins , Down-Regulation , Epithelial Cells , Epithelial-Mesenchymal Transition , Fibroblasts , Follow-Up Studies , Helicobacter pylori , Immunohistochemistry , Prospective Studies , Stomach Neoplasms , Transforming Growth Factors , Up-Regulation , Vascular Endothelial Growth Factor A , Vimentin
18.
Cancer Research and Treatment ; : 1420-1429, 2019.
Article in English | WPRIM | ID: wpr-763218

ABSTRACT

PURPOSE: MicroRNAs (miRNAs) are a group of small non-coding RNAs involved in different cancers, including lung cancer. Here, we aim to investigate the expression profiles of circulating miRNAs and their roles contributed to the progress of lung cancer. MATERIALS AND METHODS: The levels of circulating miRNA in lung cancer patients were investigated by miRNAs assay. Then we predicted the target genes of aberrantly expressing miRNAs by searching genetic databases. Based on the A549 cells transfected with miR-1246 mimics or miR-1246 inhibitor,we further measured the roles of miR-1246 involving in the epithelial-mesenchymal transition (EMT), migration and invasion capacities of lung cancer cells in vitro. Finally, we detected the effects of miR-1246 on glycogen synthase kinase-3β (GSK-3β)/β-catenin pathway by immunofluorescence and Western blot, respectively. RESULTS: We identified that 14 miRNAs were aberrantly expressed in the serum of lung cancer patients. Among them, miR-1246 was the most up-regulated. The cell assays indicated that miR-1246 significantly increased the migration and invasion capabilities of A549 lung cancer cells. Meanwhile, immunofluorescence analysis revealed that miR-1246 promoted EMT process of A549 cells accompanying with decreasing E-cadherin expression, while increasing vimentin and transforming growth factor β (TGF-β) expression. Furthermore, an online tool predicated that miR-1246 might bind to 3′-untranslated region of GSK-3β, which was confirmed by overexpression and knockdown of miR-1246 assays. CONCLUSION: Taken together, the study illustrates that miR-1246 regulates Wnt/β-catenin pathway through targeting GSK-3β/β-catenin, which partly contributing to tumor metastasis. MiR-1246 may play an essential role in the diagnosis and therapeutic of lung cancer.


Subject(s)
Humans , Blotting, Western , Cadherins , Databases, Genetic , Diagnosis , Epithelial-Mesenchymal Transition , Fluorescent Antibody Technique , Glycogen Synthase , In Vitro Techniques , Lung Neoplasms , MicroRNAs , Neoplasm Metastasis , RNA, Small Untranslated , Transforming Growth Factors , Vimentin
19.
Tuberculosis and Respiratory Diseases ; : 42-52, 2019.
Article in English | WPRIM | ID: wpr-719618

ABSTRACT

BACKGROUND: Transforming growth factor β (TGF-β), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-β1. METHODS: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-β1 and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-β1 and Smad3 were evaluated at 1 and 3 weeks. RESULTS: When A549 cells were pre-stimulated with TGF-β1 prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-β1 stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-β1 failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-β1-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-β1 and Smad3 at 1 and 3 weeks. CONCLUSION: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-β1 in vitro, and RA also decreased the expression of TGF-β1 at 1 and 3 weeks in vivo. Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-β1/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-β1-induced lung injury and fibrosis.


Subject(s)
Animals , Mice , Apoptosis , Bleomycin , Blotting, Western , Cell Differentiation , Epithelial Cells , Fibroblasts , Fibrosis , In Vitro Techniques , Lung Injury , Mitogen-Activated Protein Kinase Kinases , Mitogen-Activated Protein Kinases , Models, Animal , p38 Mitogen-Activated Protein Kinases , Phosphorylation , Protein Kinases , Smad Proteins , Transforming Growth Factor beta , Transforming Growth Factors , Tretinoin
20.
Korean Journal of Pediatrics ; : 95-101, 2019.
Article in English | WPRIM | ID: wpr-760188

ABSTRACT

PURPOSE: Increased apoptosis was recently found in the hypertrophied left ventricle of spontaneously hypertensive rats (SHRs). Although the available evidence suggests that apoptosis can be induced in cardiac cells by various insults including pressure overload, cardiac apoptosis appears to result from an exaggerated local production of angiotensin in adult SHRs. Altered expressions of Bcl associated X (Bax), Bcl-2, chemokine receptor (CCR)-2, monocyte chemoattractant protein (MCP)-1, transforming growth factor (TGF)-β1, phosphorylated extracellular signal-regulated kinases (PERK), and connexin 43 proteins, and kallikrein mRNA were investigated to explore the effects of losartan on the SHR model. METHODS: Twelve-week-old male rats were grouped as follows: control (C), SHR (hypertension: H), and losartan (L; SHRs were treated with losartan [10 mg/kg/day] for 5 weeks). Western blot and reverse transcription polymerase chain reaction assays were performed. RESULTS: Expression of Bax, CCR-2, MCP-1, TGF-β1, PERK, and connexin 43 proteins, and kallikrein mRNA was significantly increased in the H group compared to that in the C group at weeks 3 and 5. Expression of Bax, CCR-2, MCP-1, TGF-β1, and connexin 43 proteins and kallikrein mRNA was significantly decreased after losartan treatment at week 5. PERK protein expression was significantly decreased after losartan treatment at weeks 3 and 5. Bcl-2 protein expression was significantly decreased in the H group compared to that in the C group at weeks 3 and 5. CONCLUSION: Losartan treatment reduced expression of Bax, CCR-2, MCP-1, TGF-β1, PERK, and connexin 43 proteins, and kallikrein mRNA in SHRs, along with decreased inflammation and apoptosis.


Subject(s)
Adult , Animals , Humans , Male , Rats , Angiotensins , Apoptosis , Blotting, Western , Connexin 43 , Extracellular Signal-Regulated MAP Kinases , Gene Expression , Heart Ventricles , Inflammation , Kallikreins , Losartan , Monocytes , Polymerase Chain Reaction , Rats, Inbred SHR , Reverse Transcription , RNA, Messenger , Transforming Growth Factors
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