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Balanitis , Humanos , Balanitis/diagnóstico , Balanitis/terapia , Consenso , Pueblos del Este de AsiaRESUMEN
Introduction: Vitiligo, a common autoimmune acquired pigmentary skin disorder, poses challenges due to its unclear pathogenesis. Evidence suggests inflammation and metabolism's pivotal roles in its onset and progression. This study aims to elucidate the causal relationships between vitiligo and inflammatory proteins, immune cells, and metabolites, exploring bidirectional associations and potential drug targets. Methods: Mendelian Randomization (MR) analysis encompassed 4,907 plasma proteins, 91 inflammatory proteins, 731 immune cell features, and 1400 metabolites. Bioinformatics analysis included Protein-Protein Interaction (PPI) network construction, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Subnetwork discovery and hub protein identification utilized the Molecular Complex Detection (MCODE) plugin. Colocalization analysis and drug target exploration, including molecular docking validation, were performed. Results: MR analysis identified 49 proteins, 39 immune cell features, and 59 metabolites causally related to vitiligo. Bioinformatics analysis revealed significant involvement in PPI, GO enrichment, and KEGG pathways. Subnetwork analysis identified six central proteins, with Interferon Regulatory Factor 3 (IRF3) exhibiting strong colocalization evidence. Molecular docking validated Piceatannol's binding to IRF3, indicating a stable interaction. Conclusion: This study comprehensively elucidates inflammation, immune response, and metabolism's intricate involvement in vitiligo pathogenesis. Identified proteins and pathways offer potential therapeutic targets, with IRF3 emerging as a promising candidate. These findings deepen our understanding of vitiligo's etiology, informing future research and drug development endeavors.
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Condyloma acuminatum is a common sexually transmitted disease caused by human papillomavirus infection and is a benign hyperplastic lesion of the genital and perianal areas. The principle of its treatment is to remove the visible warts as much as possible and to prevent recurrence. Traditional treatment methods of condyloma acuminatum, such as CO2 laser, liquid nitrogen freezing, surgery, and topical medications, can remove warts. However, these methods have disadvantages such as pain, high recurrence rates, long treatment cycles, and scarring. Aminolevulinic acid/photodynamic therapy (ALA-PDT), a safe and effective method, has been widely used to treat condyloma acuminatum in recent years. Condyloma acuminatum occurs relatively rarely in elderly patients, in whom treatment is difficult owing to poorer physiological function. We successfully treated an 87-year-old patient with a giant condyloma acuminatum of the glans penis using six sessions of ALA-PDT at 7-day intervals and obtained satisfactory results. No recurrence was observed during a 6-month follow-up. Therefore, ALA-PDT is worth popularizing in clinical practice.
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Tumor de Buschke-Lowenstein , Condiloma Acuminado , Fotoquimioterapia , Masculino , Humanos , Anciano , Anciano de 80 o más Años , Ácido Aminolevulínico/uso terapéutico , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/uso terapéutico , Tumor de Buschke-Lowenstein/tratamiento farmacológico , Condiloma Acuminado/tratamiento farmacológico , PapillomaviridaeRESUMEN
Testosterone in male mammals is mainly secreted by testicular Leydig cells, and its secretion process is regulated by the hypothalamic-pituitary-gonadal axis. After receiving the luteinizing hormone (LH) stimulus signal, the lutropin/choriogonadotropin receptor (LHCGR) on the Leydig cell membrane transfers the signal into the cell and finally increases the secretion of testosterone by upregulating the expression of steroid hormone synthase. In previous experiments, we found that interfering with the expression of the Luman protein can significantly increase testosterone secretion in MLTC-1 cells. In this experiment, we found that knockdown of Luman in MLTC-1 cells significantly increased the concentration of cAMP and upregulated the expression of AC and LHCGR. Moreover, an analysis of the activity of the LHCGR promoter by a dual luciferase reporter system showed that knockdown of Luman increased the activity of the LHCGR promoter. Therefore, we believe that knockdown of Luman increased the activity of the LHCGR promoter and upregulated the expression of LHCGR, thereby increasing the concentration of intracellular cAMP and ultimately leading to an increase of testosterone secretion by MLTC-1 cells.
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Células Intersticiales del Testículo , Receptores de HL , Masculino , Animales , Receptores de HL/genética , Receptores de HL/metabolismo , Testosterona/metabolismo , Testículo/metabolismo , Hormona Luteinizante/farmacología , Hormona Luteinizante/metabolismo , MamíferosRESUMEN
Toxoplasma gondii (T. gondii) is a serious intracellular parasite and mammalian infection can damage the reproductive system and lead to apoptosis of Murine Leydig tumor cells (MLTC-1); however, the mechanism is unclear. The testis Leydig cell is the main testosterone synthesis cell in male mammals. We studied the mechanism of T. gondii infection on Leydig cell apoptosis in vitro. MLTC-1 were divided into control and experimental groups. Experiment group cells and tachyzoites were co-cultured, in a 1:20 ratio, for 3, 6, 9, and 12 h. T. gondii entered the cells and caused lesions at 12 h. Flow cytometry showed that the apoptosis rate of the experiment group increased with time and was significantly higher (P < 0.05) than the control group. RT-qPCR and western blot demonstrated that the expression of P53, Caspase-3, and Bax were significantly increased at 12 h (P < 0.05). Bcl-2 expression was significantly increased at 12 h (P < 0.05). The ER stress (ERS) pathway was important in cell apoptosis. RT-qPCR and western blot showed that the expression of CHOP was significantly increased at 12 h (P < 0.05). These data indicate that T. gondii induced MLTC-1 cell apoptosis may occur via the ERS pathway.
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Toxoplasma , Toxoplasmosis , Ratones , Masculino , Animales , Células Intersticiales del Testículo , Apoptosis , Técnicas de Cocultivo , MamíferosRESUMEN
Hair follicle stem cells (HFSCs) are responsible for hair growth and hair follicle regeneration. MicroRNAs have been demonstrated to be involved in the differentiation of HFSCs. Thus, this study aimed to explore the potential role of miR-149 in the differentiation of HFSCs. The isolated HFSCs were identified by flow cytometric sorting. miR-149 expression was determined during differentiation of HFSCs. Gain- and loss-of-function approaches were conducted to explore the roles of miR-149, MAPK1/ERK2, and FGF2/c-MYC in colony formation and proliferation of HFSCs. Furthermore, in vivo assays were undertaken in miR-149 knockout mice to confirm their roles in HFSC differentiation. miR-149 was found to be downregulated during HFSC differentiation, and overexpressed miR-149 restricted the proliferation and differentiation of HFSCs. miR-149 was confirmed to target and inhibit MAPK1/ERK2, which was highly expressed in and positively associated with HFSC differentiation. The MAPK1/ERK2 promotion in HFSC differentiation was achieved by augmenting expression of FGF2 and c-MYC. The in vitro effects of miR-149 were validated in in vivo experiments. Taken together, upregulated miR-149 restricted HFSC differentiation and hair growth by targeting MAPK1/ERK2 to reduce expression of FGF2 and c-MYC, which sheds light on the underlying molecular mechanism of hair growth.
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Folículo Piloso , MicroARNs , Animales , Diferenciación Celular/genética , Factor 2 de Crecimiento de Fibroblastos , Folículo Piloso/metabolismo , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Células MadreRESUMEN
BACKGROUND: Condyloma acuminatum is a benign proliferative lesion of the genitalia and perianal region caused by human papillomavirus infection, which is rare in children. Traditional treatments include topical medications, CO2 laser therapy, cryotherapy, and surgical resection, which are often associated with pain, scarring, and recurrence. In recent years, aminolevulinic acid/photodynamic therapy has been applied as a new method for treating condyloma acuminatum in adults. However, its application in young children is limited. We herein report a case of condyloma acuminatum treated by aminolevulinic acid/photodynamic therapy in a young boy. CASE REPORT: The patient was a 2-year-old boy who presented with a 10-day history of warty masses on his glans penis. Condyloma acuminatum was diagnosed based on skin lesions and histological findings. Aminolevulinic acid/photodynamic therapy was selected as the treatment method after obtaining informed consent from his guardians. We performed five rounds with a treatment interval of 7 days and achieved a complete disappearance of the skin lesions without chapping, flushing, or scarring. There was no recurrence during the 6-month follow-up period. CONCLUSION: Photodynamic therapy can be used as an alternative treatment for condyloma acuminatum in young children. More case studies are needed to confirm the value of this treatment.
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Condiloma Acuminado , Fotoquimioterapia , Adulto , Ácido Aminolevulínico/uso terapéutico , Niño , Preescolar , Cicatriz/tratamiento farmacológico , Cicatriz/patología , Condiloma Acuminado/tratamiento farmacológico , Condiloma Acuminado/patología , Humanos , Masculino , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/uso terapéuticoRESUMEN
Coccidiosis is an important parasitic disease of poultry with great economic importance. Due to drug resistance issues, the study was conducted to investigate how probiotics (Lactobacillus plantarum or L. plantarum) affected oocysts per gram of feces (OPG), fecal scores, feed conversion ratio (FCR), immunomodulatory effect in terms of the cell-mediated and humoral immune response. Serum chemistry (ALT, AST, LDH, and creatinine) was measured in different treated chicken groups. mRNA expression levels of antioxidant enzymes (SOD 1 and CAT), peptide transporter 1 (PepT 1), and tight junction proteins (ZO and CLDN 1) were also examined in chicken groups infected with Eimeria tenella (E. tenella). Chickens supplemented with L. plantarum 1 × 108 CFU (colony-forming unit) showed an improved cell-mediated and humoral immune response, compared with the control group (p < 0.05). Probiotics also enhanced the performance of antioxidant enzymes, PepT 1, and tight junction proteins, and improved serum chemistry (AST, ALT, and LDH), compared with control-infected, non-medicated chickens. However, no significant difference (p > 0.05) was observed in CLDN 1 expression level and creatinine in all treated chicken groups. These findings demonstrated that probiotics supplementation in the feed can protect the birds against E. tenella infection.
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BACKGROUND: Condyloma acuminatum (CA) is a common sexually transmitted disease caused by human papillomaviruses. Five-aminolevulinic acid-photodynamic therapy (ALA-PDT) can promote the apoptosis of keratinocytes and inhibit proliferation; however, the effect of ALA-PDT on the immune mechanism of CA tissue is not well understood. In this study, we examined the changes in Toll-like receptor 4 (TRL4) and nuclear factor kappa B (NF-κB) expression in CA tissues before ALA-PDT treatment to determine its effects and possible immune mechanisms. METHODS: Immunohistochemistry (streptavidin-peroxidase) was used to detect the expression of TLR4 and NF-κB in the keratinocytes of the patients with CA before and after ALA-PDT treatment. RESULTS: Before treatment, the positive TLR4 and NF-κB expression rates in the keratinocytes of the patients with CA were 86.53% (45/52) and 94.23% (49/52), respectively, whereas after treatment, these rates were 44.23% (23/52) and 38.46% (20/52), respectively. Positive TLR4 and NF-κB expression in the keratinocytes of CA was mostly ++ to +++ in intensity before ALA-PDT and mostly ranged from - to + after treatment. The positive expression rate and intensity of TLR4 and NF-κB in the two groups before and after ALA-PDT were significantly different (P < 0.05). There was a positive correlation between the expression of TLR4 and NF-κB in the CA tissues after ALA-PDT (r = 0.486, P < 0.05). CONCLUSIONS: ALA-PDT may relieve local immunosuppressive states by reducing TLR4 and NF-κB expression and jointly promoting CA regression, which is a potential molecular mechanism of ALA-PDT in CA treatment.
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Ácido Aminolevulínico , Fotoquimioterapia , Ácido Aminolevulínico/metabolismo , Ácido Aminolevulínico/farmacología , Ácido Aminolevulínico/uso terapéutico , Humanos , Queratinocitos , FN-kappa B/metabolismo , FN-kappa B/farmacología , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Receptor Toll-Like 4/metabolismoRESUMEN
Parasitic diseases are major trouble in many parts of the world. We consider that if a chemical can break a DNA barcode sequence, it might be used to develop a species-specific anti-parasitic agent. To examine this hypothesis, we constructed sgRNAs that target both the control (5.8S rDNA) and a DNA barcode (ITS) sequence in Eimeria tenella. In vitro experiment showed that Cas9 mRNA combined with sgRNAs could reduce the sporulation percentage of oocysts and the survival rate of sporulated oocysts and sporozoites. Quantitative real-time PCR showed that the DNAs of parasites exposed to Cas9 mRNA and sgRNAs were significantly affected, regardless of whether they were exposed to a combination of two sgRNAs or just a single sgRNA. The DNA sequencing also indicated that the experimental group exposed to two sgRNAs mixed with Cas9-induced deletion of large parts and a single sgRNA mixed with Cas9-induced mutation at sgRNA targeted fragments. In vivo trial, the effect of sgRNA and Cas9 RNA on the pathogenicity of E. tenella in chicken showed less lesion score and oocysts score (P < 0.05) in experimental groups than control groups. The results and concepts presented in this research can lead to discovering novel nucleic acid therapeutic drugs for Eimeriasis and other parasitic infections, which provide insights into the development of species-specific anti-parasitic agents.
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Eimeria tenella , Animales , Sistemas CRISPR-Cas , Pollos/genética , Eimeria tenella/genética , ARN , ARN Mensajero/genéticaRESUMEN
AIMS: The purpose of this study was to explore the precise role and mechanism of p38 inhibiting cutaneous squamous cell carcinoma associated lincRNA (PICSAR) in CSCC. MATERIALS AND METHODS: The expression levels of PICSAR, microRNA-125b (miR-125b) and yes-associated protein1 (YAP1) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation, apoptosis and invasion were evaluated by Cell Counting Kit-8 (CCK-8) assay, flow cytometry, transwell assay, respectively. The interaction between miR-125b and PICSAR or YAP1 was predicted by bioinformatics software and confirmed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Western blot was employed to detect the protein expression of YAP1. The mice xenograft model was established to investigate the role of PICSAR in vivo. KEY FINDINGS: PICSAR was upregulated in CSCC tissues and cells. PICSAR knockdown inhibited cell proliferation and invasion and induced apoptosis in CSCC cells. Moreover, miR-125b could directly bind to PICSAR and its inhibition reversed the effect of PICSAR knockdown on proliferation, invasion and apoptosis in CSCC cells. In addition, YAP1 was a direct target of miR-125b and its overexpression attenuated the anti-cancer role of miR-125b in CSCC cells. Furthermore, YAP1 expression was positively regulated by PICSAR and negatively regulated by miR-125b. Besides, interference of PICSAR suppressed tumor growth by upregulating miR-125b and downregulating YAP1. SIGNIFICANCE: PICSAR knockdown suppressed cell proliferation and invasion and promoted apoptosis in CSCC cells by regulating miR-125b/YAP1 axis, providing new sights for treatment of CSCC.
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Proteínas Adaptadoras Transductoras de Señales/metabolismo , Biomarcadores de Tumor/metabolismo , Carcinoma de Células Escamosas/patología , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , ARN Largo no Codificante/genética , Neoplasias Cutáneas/patología , Factores de Transcripción/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Apoptosis , Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Movimiento Celular , Proliferación Celular , Progresión de la Enfermedad , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Pronóstico , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/metabolismo , Factores de Transcripción/genética , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de Xenoinjerto , Proteínas Señalizadoras YAPAsunto(s)
Mano , Nevo Pigmentado/patología , Neoplasias Cutáneas/patología , Adulto , Femenino , HumanosRESUMEN
BACKGROUND: Dysfunction of the DNA methylation was associated with stem cell reprogramming. Moreover, DNA methyltransferase 1 (DNMT1) deficiency was involved in the differentiation of hair follicle stem cell (HFSc), but the molecular mechanisms remain unknown. METHODS: HFSc from human scalp tissues were isolated and cultured. The oil red O staining was used to observe the adipogenesis. The interaction relationship between microRNA (miR)-214-3p and mitogen-activated protein kinase 1 (MAPK1) was accessed by dual-luciferase reporter gene assay. The methylation level of miR-214-3p promoter was detected by methylation-specific PCR and the enrichment of DNMT1 in miR-214-3p promoter by chromatin immunoprecipitation assay. A mouse model of trauma was established to observe the skin regeneration at 0, 6, and 14 days. RESULTS: Expression of DNMT1 and MAPK1 was increased in the HFSc, while the expression of miR-214-3p was reduced. Moreover, DNMT1 inhibited the expression of miR-214-3p by promoting the promoter methylation of miR-214-3p. Overexpression of DNMT1 could reduce the expression of miR-214-3p, but increase the expression of MAPK1 and the extent of extracellular signal regulated kinase (ERK)1/2 phosphorylation, leading to enhanced adipogenic differentiation. Importantly, DNMT1 promoted skin regeneration in vivo. Conversely, overexpression of miR-214-3p could reverse the effects of DNMT1 on adipogenesis of HFSc. CONCLUSION: DNMT1 promotes adipogenesis of HFSc by mediating miR-214-3p/MAPK1/p-ERK1/2 axis. This study may provide novel biomarkers for the potential application in stem cell therapy.
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Adipogénesis , ADN (Citosina-5-)-Metiltransferasa 1/metabolismo , Folículo Piloso/citología , MicroARNs , Células Madre/citología , Adipogénesis/genética , Diferenciación Celular , Células Cultivadas , Metilación de ADN , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Células Madre/metabolismoRESUMEN
Hair follicle stem cells (HFSCs) contribute to the regeneration of hair follicles (HFs), thus accelerating hair growth. microRNAs (miRs) are potential regulators in various cellular processes, including HFSC proliferation and differentiation. This study proposed a potential target, enhancer of zeste homolog 2 (EZH2) for facilitating hair growth, due to its function over HFSC activities by mediating the miR-22/serine/threonine kinase 40 (STK40)/myocyte enhancer factor 2 (MEF2)/alkaline phosphatase (ALP) axis. Gain- and loss-of-function approaches were adopted to explore the roles of EZH2, miR-22, and STK40 in the proliferation and apoptosis of HFSCs, along with the functional relevance of MEF2-ALP activity. STK40 was elevated during HFSC differentiation, which was found to facilitate HFSC proliferation, but impede their apoptosis by activating MEF2-ALP. Mechanically, miR-22 targeted and inversely regulated STK40, which inhibited MEF2-ALP activity to impede HFSC proliferation and differentiation. Moreover, EZH2 elevated the STK40 expression by repressing miR-22 to promote the proliferation and differentiation of HFSCs. Furthermore, in vivo experiments further validated the roles of EZH2 and STK40 on hair follicle neogenesis and hair growth. Collectively, EZH2 elevated the STK40 expression by downregulating miR-22, consequently accelerating differentiation of HFSCs and hair growth, which sheds light on the underlying molecular mechanism responsible for hair growth.
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Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Folículo Piloso/citología , MicroARNs/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Diferenciación Celular/genética , Células Cultivadas , Proteína Potenciadora del Homólogo Zeste 2/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , MicroARNs/genética , Proteínas Serina-Treonina Quinasas/genética , Células Madre/metabolismoRESUMEN
Condyloma acuminata are a type of verrucous hyperplasia in the genital and perianal areas caused by human papilloma virus (HPV) infection. Traditional treatment methods for condyloma acuminata, such as topical medications, carbon dioxide (CO2) laser, cryotherapy, and surgical excision, can be effective at removing warts, however, they do not eliminate subclinical and latent HPV infection, resulting in a high recurrence rate and even leaving trauma and scars. We report a case of condyloma acuminata covering the glans penis, considering our patient had a singular, large lesion, thus, to reduce the risk of recurrence and minimize the trauma caused by treatment, we chose ALA/photodynamic therapy (ALA-PDT). We provided eight rounds of ALA-PDT at a treatment interval of 7 days, the skin lesion disappeared completely, leaving a chapped, flushed glans without scarring. There was no recurrence during the 6-month follow-up period. Thus, we firmly believe that ALA-PDT is an effective, safe, and curative treatment, and is worth popularizing in clinical practice.
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Condiloma Acuminado , Fotoquimioterapia , Ácido Aminolevulínico/uso terapéutico , Condiloma Acuminado/tratamiento farmacológico , Humanos , Masculino , Pene , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/uso terapéuticoRESUMEN
Fibroblast growth factor (FGF2) is reported to affect the proliferation, differentiation, and survival abilities of stem cells. In this study, we hypothesize that FGF2 might promote the differentiation of hair follicle stem cell (HFSCs) into endothelial cells (ECs), in a manner dependent on STAT5 activation. We first treated human HFSCs with recombinant human FGF2 to determine the involvement of FGF2 in the differentiation of HFSCs. Then the expression of EC-specific markers including von Willebrand factor (vWF), VE-cadherin, CD31, FLT-1, KDR and Tie2 was evaluated using immunofluorescence and flow cytometry, while the expression of HFSC-specific markers such as K15, K19, Lgr5, Sox9 and Lhx2 was determined by flow cytometry. Next, in vitro tube formation was performed to confirm the function of FGF2, and low-density lipoprotein (LDL) uptake by ECs and HFSCs was studied by Dil-acetylated LDL assay. In addition, we transduced FGF2-treated HFSCs with constitutive-active or dominant-negative STAT5A adenovirus vectors. FGF2 up-regulated the expression of EC-specific markers, and promoted the differentiation of HFSCs into ECs, tube formation and LDL uptake. The phosphorylated STAT5 was translocated into the nucleus of HFSCs after FGF2 treatment, but this translocation was blocked by the dominant-negative STAT5A mutant. FGF2 increased the differentiation potential through the activation of STAT5 in vivo. Taken together, we find that FGF2 promotes the differentiation of HFSCs into ECs via activated STAT5, which gives a new perspective on the role of FGF2 in the development of ischemic vascular disease.
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Diferenciación Celular , Células Endoteliales/citología , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Folículo Piloso/citología , Factor de Transcripción STAT5/metabolismo , Células Madre/citología , Animales , Células Cultivadas , Células Endoteliales/metabolismo , Factor 2 de Crecimiento de Fibroblastos/genética , Folículo Piloso/metabolismo , Humanos , Factor de Transcripción STAT5/genética , Células Madre/metabolismoAsunto(s)
Vesícula/diagnóstico , Pénfigo/diagnóstico , Diagnóstico Diferencial , Humanos , Recién Nacido , MasculinoRESUMEN
MicroRNAs (miRNAs) have been widely accepted to play important roles in the regulation of keratinocyte functions. Here, we aimed to further explore the role and underlying mechanism of miR-125b-5p and miR-181b-5p in psoriasis. The expression levels of miR-125b-5p, miR-181b-5p and Akt3 mRNA were detected by qRT-PCR assay. Cell proliferation ability was determined by MTT assay and BrdU incorporation assay. Dual-luciferase reporter assay and RNA Immunoprecipitation assay were used to confirm the targeted interaction between miR-125b-5p or miR-181b-5p and Akt3 in human epidermal keratinocytes (HEKs). The levels of ki-67, Akt3 protein, Akt, p-Akt, mTOR and p-mTOR were measured by Western blot. Our study indicated that miR-125b-5p and miR-181b-5p were downregulated (about 61.3% with miR-125b-5p and 60.4% with miR-181b-5p) and Akt3 was upregulated (about 2.68-fold) in psoriasis. Upregulation of miR-125b-5p or miR-181b-5p resulted in about a 33% or 40% reduction of HEKs proliferation in vitro, while Akt3 overexpression triggered a 1.3-fold enhancement on HEKs proliferation. Akt3 was a direct target of miR-125b-5p or miR-181b-5p. Moreover, HEKs proliferation ability in cotransfection of miR-125b-5p mimics (or miR-181-5p mimics) and vector-Akt3 group was about 2-fold (or 1.98-fold) that in the miR-125b-5p mimics (or miR-181-5p mimics) alone group. Akt/mTOR signaling was involved in miR-125b-5p mimics- or miR-181b-5p mimics-mediated inhibition effect on HEKs proliferation. Our study suggested that the upregulation of miR-125b-5p or miR-181b-5p inhibited HEKs proliferation at least partly by targeting Akt3, providing novel mechanisms of miRNAs involved in psoriasis.
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Proliferación Celular/genética , MicroARNs/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Psoriasis/genética , Antagomirs/farmacología , Biopsia , Estudios de Casos y Controles , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Regulación hacia Abajo , Voluntarios Sanos , Humanos , Queratinocitos , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , Cultivo Primario de Células , Proteínas Proto-Oncogénicas c-akt/metabolismo , Psoriasis/patología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Piel/citología , Piel/patología , Serina-Treonina Quinasas TOR/metabolismo , Regulación hacia ArribaRESUMEN
The proliferation and differentiation of hair follicle stem cells (HFSCs) is regulated by several signaling pathways, including BMP and PTEN. Therefore, this study intended to clarify the potential effects of two such regulators, BMP2 and PTEN, on HFSC differentiation. HFSCs were subjected to BMP2, noggin (BMP2 ligand inhibitor), rapamycin (Rapa, autophagy inducer), 3-methyladenine (3-MA, autophagy inhibitor), or shRNA against PTEN. The differentiation of HFSCs was evaluated using oil red O staining and autophagy was assessed using the transmission electron microscope. Then expression of epidermal differentiation marker (K10 and involucrin), adipogenic markers (PPAR-γ2, aP2, perilipin2, and Adipoq), keratinocyte-specific marker (K15), proliferation-related markers (PCNA and Ki67) and autophagy-related factors (Atg5, Atg7, Atg12, Beclin-1 and LC3-II/LC3-I) was examined by RT-qPCR and Western blot analysis. Next, HFSCs were treated with 3-MA, or shRNA against Atg5 or Atg7 to verify the effect of autophagy on differentiation of BMP2-treated HFSCs. Finally, the effect of BMP2 on HFSC differentiation was verified by a mouse wound model. HFSCs overexpressing BMP2 exhibited elevated expression of epidermal differentiation marker, adipogenic markers and autophagy-related factors but inhibited expression of keratinocyte-specific marker and proliferation-related markers. Furthermore, we found that PTEN promoted the differentiation of BMP2-treated HFSCs by inducing autophagy. In vivo experiments further confirmed the roles of BMP2/PTEN on differentiation of HFSCs. Taken together, BMP2 up-regulated PTEN and consequently induced autophagy to facilitate HFSC differentiation.