RESUMEN
BACKGROUND: Thin-cap fibroatheroma is a rupture-prone vulnerable plaque that leads to acute coronary syndrome (ACS). However, its underlying mechanisms are not fully understood. Several studies have investigated the clinical association between angiopoietin-like protein 4 (ANGPTL4) and coronary artery disease. Therefore, this study aimed to investigate the correlation of plasma ANGPTL4 in culprit lesion of ACS patients using intravascular ultrasound (IVUS) and virtual-histology IVUS (VH-IVUS). METHODS: Fifty patients newly diagnosed with ACS between March to September 2021 were selected. Blood samples for baseline laboratory tests, including ANGPTL4, were collected before percutaneous coronary intervention (PCI), and all pre- and post-PCI IVUS examinations were performed of the culprit lesions. RESULTS: Linear regression analysis between plasma ANGPTL4 and grayscale IVUS/VH-IVUS parameters revealed that plasma ANGPTL4 was strongly correlated with the necrotic core (NC) of the minimal lumen site (r = -0.666, p = 0.003) and largest NC site (r = -0.687, p < 0.001), and patients with lower plasma ANGPTL4 levels showed a significantly higher proportion of TFCA. CONCLUSION: The present study further demonstrated the protective role of ANGPTL4 in the spectrum of atherosclerotic development in patients with ACS by culprit lesion morphology analysis using IVUS and VH-IVUS.
Asunto(s)
Síndrome Coronario Agudo , Enfermedad de la Arteria Coronaria , Intervención Coronaria Percutánea , Placa Aterosclerótica , Humanos , Síndrome Coronario Agudo/diagnóstico por imagen , Síndrome Coronario Agudo/patología , Angiopoyetinas , Angiografía Coronaria , Enfermedad de la Arteria Coronaria/patología , Vasos Coronarios/diagnóstico por imagen , Necrosis/patología , Placa Aterosclerótica/diagnóstico por imagen , Placa Aterosclerótica/patología , Ultrasonografía IntervencionalRESUMEN
Atherosclerosis, the leading cause of death, is a vascular disease of chronic inflammation. We recently showed that angiopoietin-like 4 (ANGPTL4) promotes cardiac repair by suppressing pathological inflammation. Given the fundamental contribution of inflammation to atherosclerosis, we assessed the role of ANGPTL4 in the development of atherosclerosis and determined whether ANGPTL4 regulates atherosclerotic plaque stability. We injected ANGPTL4 protein twice a week into atherosclerotic Apoe-/- mice and analyzed the atherosclerotic lesion size, inflammation, and plaque stability. In atherosclerotic mice, ANGPTL4 reduced atherosclerotic plaque size and vascular inflammation. In the atherosclerotic lesions and fibrous caps, the number of α-SMA(+), SM22α(+), and SM-MHC(+) cells was higher, while the number of CD68(+) and Mac2(+) cells was lower in the ANGPTL4 group. Most importantly, the fibrous cap was significantly thicker in the ANGPTL4 group than in the control group. Smooth muscle cells (SMCs) isolated from atherosclerotic aortas showed significantly increased expression of CD68 and Krüppel-like factor 4 (KLF4), a modulator of the vascular SMC phenotype, along with downregulation of α-SMA, and these changes were attenuated by ANGPTL4 treatment. Furthermore, ANGPTL4 reduced TNFα-induced NADPH oxidase 1 (NOX1), a major source of reactive oxygen species, resulting in the attenuation of KLF4-mediated SMC phenotypic changes. We showed that acute myocardial infarction (AMI) patients with higher levels of ANGPTL4 had fewer vascular events than AMI patients with lower levels of ANGPTL4 (p < 0.05). Our results reveal that ANGPTL4 treatment inhibits atherogenesis and suggest that targeting vascular stability and inflammation may serve as a novel therapeutic strategy to prevent and treat atherosclerosis. Even more importantly, ANGPTL4 treatment inhibited the phenotypic changes of SMCs into macrophage-like cells by downregulating NOX1 activation of KLF4, leading to the formation of more stable plaques.
Asunto(s)
Aterosclerosis , Placa Aterosclerótica , Ratones , Animales , Placa Aterosclerótica/patología , Factor 4 Similar a Kruppel , Músculo Liso Vascular , Regulación hacia Abajo , Ratones Noqueados para ApoE , Aterosclerosis/patología , Fenotipo , Miocitos del Músculo Liso/metabolismo , Inflamación/metabolismo , Ratones Endogámicos C57BL , Células CultivadasRESUMEN
Resistin-like alpha (Retnla) is a member of the resistin family and known to modulate fibrosis and inflammation. Here, we investigated the role of Retnla in the cardiac injury model. Myocardial infarction (MI) was induced in wild type (WT), Retnla knockout (KO), and Retnla transgenic (TG) mice. Cardiac function was assessed by echocardiography and was significantly preserved in the KO mice, while worsened in the TG group. Angiogenesis was substantially increased in the KO mice, and cardiomyocyte apoptosis was markedly suppressed in the KO mice. By Retnla treatment, the expression of p21 and the ratio of Bax to Bcl2 were increased in cardiomyocytes, while decreased in cardiac fibroblasts. Interestingly, the numbers of cardiac macrophages and unsorted bone marrow cells (UBCs) were higher in the KO mice than in the WT mice. Besides, phosphorylated histone H3(+) cells were more frequent in bone marrow of KO mice. Moreover, adiponectin in UBCs was notably higher in the KO mice compared with WT mice. In an adoptive transfer study, UBCs were isolated from KO mice to transplant to the WT infarcted heart. Cardiac function was better in the KO-UBCs transplanted group in the WT-UBCs transplanted group. Taken together, proliferative and adiponectin-rich bone marrow niche was associated with substantial cardiac recovery by suppression of cardiac apoptosis and proliferation of cardiac fibroblast.
Asunto(s)
Adipoquinas/metabolismo , Células de la Médula Ósea/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Infarto del Miocardio/fisiopatología , Animales , Apoptosis , Masculino , RatonesRESUMEN
Mesenchymal stem cells (MSCs) can suppress pathological inflammation. However, the mechanisms underlying the association between MSCs and inflammation remain unclear. Under coculture conditions with macrophages, MSCs highly expressed angiopoietin-like 4 (ANGPTL4) to blunt the polarization of macrophages toward the proinflammatory phenotype. ANGPTL4-deficient MSCs failed to inhibit the inflammatory macrophage phenotype. In inflammation-related animal models, the injection of coculture medium or ANGPTL4 protein increased the antiinflammatory macrophages in both peritonitis and myocardial infarction. In particular, cardiac function and pathology were markedly improved by ANGPTL4 treatment. We found that retinoic acid-related orphan receptor α (RORα) was increased by inflammatory mediators, such as IL-1ß, and bound to ANGPTL4 promoter in MSCs. Collectively, RORα-mediated ANGPTL4 induction was shown to contribute to the antiinflammatory activity of MSCs against macrophages under pathological conditions. This study suggests that the capability of ANGPTL4 to induce tissue repair is a promising opportunity for safe stem cell-free regeneration therapy from a translational perspective.
Asunto(s)
Proteína 4 Similar a la Angiopoyetina/fisiología , Activación de Macrófagos , Macrófagos/metabolismo , Células Madre Mesenquimatosas/metabolismo , Infarto del Miocardio/terapia , Peritonitis/terapia , Proteína 4 Similar a la Angiopoyetina/genética , Proteína 4 Similar a la Angiopoyetina/metabolismo , Animales , Antiinflamatorios no Esteroideos , Polaridad Celular , Células Cultivadas , Técnicas de Cocultivo , Modelos Animales de Enfermedad , Humanos , Inflamación/inmunología , Inflamación/terapia , Mediadores de Inflamación/metabolismo , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Infarto del Miocardio/inmunología , Miocarditis/etiología , Miocarditis/prevención & control , Miembro 1 del Grupo F de la Subfamilia 1 de Receptores Nucleares/metabolismo , Peritonitis/inmunología , Receptores de Ácido Retinoico/metabolismoRESUMEN
Bone marrow-derived mesenchymal stem cells (BMMSCs) are used extensively for cardiac repair and interact with immune cells in the damaged heart. Macrophages are known to be modulated by stem cells, and we hypothesized that priming macrophages with BMMSCs would enhance their therapeutic efficacy. Rat bone marrow-derived macrophages (BMDMs) were stimulated by lipopolysaccharide (LPS) with or without coculture with rat BMCs. In the LPS-stimulated BMDMs, induction of the inflammatory marker iNOS was attenuated, and the anti-inflammatory marker Arg1 was markedly upregulated by coculture with BMMSCs. Myocardial infarction (MI) was induced in rats. One group was injected with BMMSCs, and a second group was injected with MIX (a mixture of BMMSCs and BMDMs after coculture). The reduction in cardiac fibrosis was greater in the MIX group than in the BMC group. Cardiac function was improved in the BMMSC group and was substantially improved in the MIX group. Angiogenesis was better in the MIX group, and anti-inflammatory macrophages were more abundant in the MIX group than in the BMMSC group. In the BMMSCs, interferon regulatory factor 5 (IRF5) was exclusively induced by coculture with macrophages. IRF5 knockdown in BMMSCs failed to suppress inflammatory marker induction in the macrophages. In this study, we demonstrated the successful application of BMDMs primed with BMMSCs as an adjuvant to cell therapy for cardiac repair.
Asunto(s)
Macrófagos/metabolismo , Trasplante de Células Madre Mesenquimatosas/métodos , Infarto del Miocardio/terapia , Animales , Arginasa/genética , Arginasa/metabolismo , Línea Celular , Células Cultivadas , Medios de Cultivo Condicionados/farmacología , Humanos , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Ratones , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Although cell therapy is emerged for cardiac repair, its efficacy is modest by intracoronary infusion. Therefore, we established the intramyocardial delivery technique using a left ventricular (LV) mapping system (NOGA® XP) using 18 pigs. After adipose tissue-derived mesenchymal stem cells (ATSCs) were delivered intramyocardially to porcine infarcted heart, LV ejection fraction (EF) was increased, and LV chamber size was decreased. We proved the therapeutic effect of intramyocardial injection of ATSC through a LV mapping system in the porcine model for the first time in Korea. The adoption of this technique may accelerate the translation into a clinical application in the near future.
Asunto(s)
Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Infarto del Miocardio/terapia , Animales , Modelos Animales de Enfermedad , Vías de Administración de Medicamentos , Ecocardiografía , Corazón/diagnóstico por imagen , Infarto del Miocardio/diagnóstico por imagen , República de Corea , Porcinos , Función Ventricular Izquierda/fisiologíaRESUMEN
Embryonic stem (ES) cells can be valuable for monitoring differentiation processes and for improving applications in basic developmental biology. The application of ES cells can be a useful tool for drug discovery and toxicology. Therefore, we suggest the high-throughput screening (HTS) system based on ES cells in this study. Firstly, we optimized the feeder-free condition and seeding cell number which can maintained for at least 7 days without over-confluency. We analyzed the system by cell viability, proliferation activity, RT-PCR and morphologic/immunohistochemical evaluations. The optimal cell seeding number was 30/well that was maintained the typical colonial morphology over 9 d with 1,000 U/ml LIF in the limited space. The cell in optimized condition expressed ALP, SSEA-1, Oct 4 and Nanog and the genetic expressions showed similar to protein expressions. The cell lineage marker expressions showed faint or none. The cell viability and proliferation activity were increased in time-dependent manner in our optimized HTS system. In conclusion, the novel HTS system using ES cells can by useful for developing models for drug discovery as well as toxicological screening in the near future.
Asunto(s)
Células Madre Embrionarias/metabolismo , Ensayos Analíticos de Alto Rendimiento/métodos , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Biomarcadores/metabolismo , Diferenciación Celular , Linaje de la Célula , Células Madre Embrionarias/citología , Proteínas de Homeodominio/metabolismo , Antígeno Lewis X/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Proteína Homeótica Nanog , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: To evaluate the potential of human umbilical cord-derived stromal cells (hUCSCs) as a human feeder for human embryonic stem cells (ESCs). DESIGN: Prospective study. SETTING: Laboratory of Molecular Genetics and Stem Cell Differentiation, Dental Research Institute, School of Dentistry, Seoul National University. INTERVENTION(S): The hUCSCs were established, and human ESCs were cultured on established hUCSCs without serum. MAIN OUTCOME MEASURE(S): Cell-surface markers, karyotyping, and teratoma formation. RESULT(S): Primary cultures of hUCSCs from individual umbilical cords were maintained by an established protocol. Human ESCs on hUCSC layers were successfully maintained in serum-free culture medium past passage 30. Compared with hESCs on mouse feeder cells, the hESCs on hUCSCs showed similar levels of pluripotency-related cell-surface markers, self-renewal capacity, and teratoma formation in immune-deficient mice. These ESCs cultured on hUCSCs had a normal karyotype, even after long-term culture. CONCLUSION(S): The hUCSCs supported self-renewal of hESCs in serum-free conditions. This culture system has the potential to facilitate the development of clinical-grade hESCs for regenerative medicine.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Células Madre Embrionarias/citología , Cordón Umbilical/citología , Inestabilidad Cromosómica , Medio de Cultivo Libre de Suero , Células Madre Embrionarias/metabolismo , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Humanos , Células Madre Pluripotentes/citología , Células del EstromaRESUMEN
Application of human embryonic stem cells (hESCs) to stem-cell therapy is not feasible because of the risk of tumorigenicity and rejection. In contrast, human mesenchymal stem cells (hMSCs) are free from the risk of tumorigenicity and also have immune privilege. However, hMSCs obtained from adults have infinite variety in terms of the biological characteristics and functionality. We report here a new derivation method of hMSCs from hESCs. The derivation of hMSCs from three different hESC lines (SNUhES3, CHA3-hESC, and H9) was performed by embryoid bodies formation and subsequent culture with stage-different media without using inductive xenogenic feeder and mechanical selection procedure. The derived cells were morphologically similar to the unique fingerprint-like pattern of hMSCs and grew stably for at least 35 passages in vitro. These cells had hMSCs-like immunophenotypes: negative for CD34 and CD45; positive for CD29, CD44, CD73, CD90, and CD105. They could be differentiated into multiple lineages including osteocytes, chondrocytes, adipocytes, and myocytes. They maintained normal karyotype during the long-term cultivation and did not show tumorigenicity when transplanted into the immunodeficient mice. In conclusion, the new embryoid body-based derivation method of hMSCs from hESCs is simple, safe, and reproducible in three different hESC lines. We expect that this method will provide a more effective and powerful tool to derive hMSCs from various hESC lines.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Embrión de Mamíferos/citología , Células Madre Embrionarias/citología , Células Madre Mesenquimatosas/citología , Adulto , Animales , Diferenciación Celular , Línea Celular , Proliferación Celular , Forma de la Célula , Electrocardiografía , Células Madre Embrionarias/metabolismo , Estudios de Factibilidad , Citometría de Flujo , Humanos , Cariotipificación , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Ratones , Miocardio/patologíaRESUMEN
Mouse two-cell embryos of block (ICR) or nonblock (F1 of C57BL x DBA) strains were cultured in modified Chatot, Ziomek, and Bavister medium, supplemented with bovine serum albumin or polyvinyl alcohol, a synthetic macromolecule. The supplementation did not influence morula compaction and blastocyst formation, mean cell numbers of total blastomeres, inner cell mass (ICM) cells and trophectodermal (TE) cells in blastocysts, rates of pregnancy and delivery, mean litter size, number of implantation spots in pregnant females, and the expression of several genes related to pluripotency, organogenesis, and implantation.
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Medios de Cultivo/química , Medios de Cultivo/metabolismo , Desarrollo Embrionario/fisiología , Desarrollo Fetal/fisiología , Técnicas de Cultivo de Órganos/métodos , Animales , Estudios de Factibilidad , Femenino , Ratones , Ratones Endogámicos ICR , Proteínas/química , Proteínas/metabolismoRESUMEN
OBJECTIVE: To assess whether embryonic stem (ES) cells could be derived from the aggregation of diploid cells with tetraploid embryos. DESIGN: Randomized, prospective study. SETTING: University embryology and gamete biotechnology laboratory. ANIMAL(S): F1 (C57BL6/DBA2) mice. INTERVENTION(S): Four- to eight-cell F1 tetraploid embryos were aggregated with 10 to 15 donor E14 ES cells. MAIN OUTCOME MEASURE(S): Embryogenesis and ES cell establishment. RESULT(S): No difference (78% to 89%) in blastocyst formation was detected between the aggregated tetraploid and the control diploid embryos. In a total of 27 transfers, pregnancy was detected in three tetraploid (23.1%) and five diploid (35.7%) cases, and three live births developed from the aggregated tetraploid embryos. The tetraploid blastocysts without aggregation were plated, but no ES cell-like colony was formed. Six of eight aggregated blastocysts derived well-proliferated colonies, which were positive for anti-stage-specific embryonic antigen (SSEA)-1 antibody, Oct-4, and alkaline phosphatase. The microsatellite assay confirmed the homogenous makeup among the donor E14 cells and live-birth and ES-like cells derived from the E14-aggregated, tetraploid embryo. CONCLUSION(S): The aggregation of pluripotent diploid cells with tetraploid embryos yielded live births and ES-like cells that were homogenous to the donor diploid cells.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Células Clonales/citología , Clonación de Organismos/métodos , Diploidia , Embrión de Mamíferos/citología , Células Madre Pluripotentes/citología , Poliploidía , Animales , Agregación Celular/fisiología , Células Cultivadas , Células Clonales/fisiología , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Células Madre Pluripotentes/fisiología , Ingeniería de Tejidos/métodosRESUMEN
This study was conducted to evaluate whether refining mineral oil and the addition of hemoglobin and/or glucose to a serum-free medium could improve in vitro-development of embryos cultured in a chemically semi-defined microdroplet culture system. Block strain, outbred (ICR) mouse 1- or 2-cell embryos were cultured in 5 microl droplets of Chatot, Ziomek and Bavister medium overlaid with mineral oil of different types, and preimplantation development to the blastocyst stage was subsequently monitored. In the experiment 1, either Sigma (M-8410) or BDH (GPR) mineral oil with or without washing was used for embryo culture and, distilled water (DW) or culture medium was used as a washing agent. As results, better (P<0.0001) development of 1-cell embryos was found in the Sigma than in the BDH; more blastocysts developed in Sigma oil washed with culture medium than in the others (37% vs. 0%). Subsequently, 1- (experiment 2) or 2-cell (experiment 3) embryos were cultured in the droplets overlaid with medium-washed Sigma oil, to which 0.001 mg/ml hemoglobin and/or 5.6 mM glucose were supplemented at the 1-cell and the 4-cell stages, respectively. Regardless of embryo stages, blastocyst formation was significantly improved by the addition of hemoglobin (54 to 48% vs. 42 to 31% in 1-cell and 83 to 78% vs. 65 to 68% in 2-cell embryos) and this effect was independent of glucose addition. In conclusion, the selection and washing of mineral oil, and the addition of hemoglobin is beneficial for improving the efficacy of a drop embryo culture system using a serum-free medium.
Asunto(s)
Embrión de Mamíferos/citología , Glucosa/farmacología , Hemoglobinas/farmacología , Animales , División Celular/efectos de los fármacos , Medio de Cultivo Libre de Suero , Embrión de Mamíferos/efectos de los fármacos , Embrión de Mamíferos/fisiología , Desarrollo Embrionario y Fetal/efectos de los fármacos , Fertilización In Vitro/métodos , Ratones , Ratones Endogámicos ICR , Aceite Mineral , Técnicas de Cultivo de Órganos/métodosRESUMEN
PURPOSE: This study was to investigate the effects of problem solving group counseling on the index of obesity and health habits for obese children. METHOD: Forty seven obese children participated in the study(Exp.=22, Cont.=25). Children were recruited from the forth and fifth grade with higher than 20% of the obesity degree. The problem solving counseling lasted for 10 weeks. In order to evaluate the effects of counseling, physical characteristics and health habits were measured three times; pretest, posttest, and at 10 weeks follow-up. The obtained data was analyzed by chi(2)-test, t-test, and repeated measures ANOVA, using the SPSS WIN 10.0 program. RESULT: Problem solving group counseling was effective on the physical characteristics(BMI, obesity degree, body fat ratio, waist measurement) and health habits over time. Children in the experimental group controlled their body weight better and reported lower scores in the index of obesity than children in the control group at 10 weeks follow-up. CONCLUSION: This counseling program helped obese children modify their health habits so that they could decrease their scores in the obesity index. It can be concluded that problem-solving counseling enhanced problem-solving abilities of obese children, which could help modify their ordinary health habits.