RESUMEN
The objective of this study was to observe the acute cytotoxic effects of hematoporphyrin monomethyl ether sonodynamic therapy (HMME-SDT) on hypertrophic scar fibroblasts of rabbit ears. We first assessed the effects of different irradiation times and HMME concentrations on the survival of hypertrophic scar fibroblasts using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to determine the optimum irradiation time and HMME concentration. The hypertrophic scar fibroblast cell suspensions of the rabbit ears were divided into four groups, the survival rates were detected using the MTT assay, and the type of cell death was detected by Annexin V/propidium iodide (PI) double staining flow cytometry. Our results showed that HMME-SDT significantly reduced the viability of hypertrophic scar fibroblasts of rabbit ears at ultrasonic irradiation times of 30, 60, and 90 s, but not 10 s (P < 0.05). HMME alone had no significant effect on the cell survival rate at any irradiation time (P > 0.05). In contrast, the cell survival rate was significantly decreased at an irradiation time of 10 s and HMME concentrations of 20 and 50 µg/mL (P < 0.05). Furthermore, Annexin V/PI double staining showed necrosis and apoptosis of the hypertrophic scar fibroblasts. Given our results, HMME might be an effective sound-sensitive agent for SDT as it has a significant lethal effect on hypertrophic scar fibroblasts of rabbit ear cultured in vitro. HMME-SDT may therefore provide a new method for the treatment of hypertrophic scar formation.
Asunto(s)
Cicatriz Hipertrófica/diagnóstico por imagen , Oído/diagnóstico por imagen , Oído/patología , Hematoporfirinas/uso terapéutico , Terapia por Ultrasonido/métodos , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cicatriz Hipertrófica/cirugía , Fibroblastos/diagnóstico por imagen , Fibroblastos/patología , Conejos , Distribución Aleatoria , Especies Reactivas de Oxígeno/uso terapéutico , UltrasonografíaRESUMEN
Hereditary deficiency of factor VIII (FVIII) leads to hemophilia A, a severe X-linked bleeding disorder. Current therapies include fixed-dose FVIII prophylaxis, factor replacement therapy, and most recently, gene therapy. Prophylaxis and FVIII replacement therapies are limited by incomplete efficacy, high cost, restricted availability, and development of neutralizing antibodies in chronically treated individuals. Limited success has been obtained in preclinical trials using gene therapy for the treatment of hemophilia. Therefore, new options for therapy for hemophilia A are needed. We evaluated the potential of embryonic stem cells for correcting hemophilia A in mice. FVIII-deficient mouse blastocysts were collected and injected with mouse embryonic stem cells stably expressing green-fluorescent protein (GFP) and transferred to pseudopregnant recipient mice. Expression of FVIII was measured in the liver and plasma of the 5 chimeric mice that were produced. Three of these mice were GFP-positive at the age of 6 months. The plasma FVIII activity levels were equal to those of wild-type mice. These data demonstrate that embryonic stem cell transplantation at an early embryonic stage has potential as therapy for this progressively debilitating, life-threatening bleeding disorder.
Asunto(s)
Células Madre Embrionarias/citología , Factor VIII/metabolismo , Hemofilia A/terapia , Trasplante de Células Madre , Animales , Quimera , Células Madre Embrionarias/metabolismo , Factor VIII/genética , Femenino , Proteínas Fluorescentes Verdes/metabolismo , Hemofilia A/sangre , Humanos , Inmunohistoquímica , Hígado/metabolismo , Hígado/patología , Ratones , Ratones Transgénicos , Microscopía Fluorescente , FenotipoRESUMEN
Zinc finger protein 191, ZNF24 and Zfp191 in both humans and mice belong to the SCAN domain subfamily of Krüppel-like zinc finger transcription factors. Previous studies have suggested that Zfp191 is a pleiotropic factor involved in embryonic development, hematopoiesis and tumorigenesis. However, little is known about its target genes or its role in other physiological and pathological processes. We have identified the putative target genes of Zfp191, using an in silico genome-wide scan. Three hundred and fifty-five putative target genes were identified, which were enriched into the pathways of immune response according to the pathway analysis. These targets indicated that Zfp191 may function as a mediator of the immune response. This was verified in mice heterozygous for Zfp191 (Zfp191(+/-)) using a lipopolysaccharide (LPS)-induced endotoxic shock model. After LPS injection, Zfp191(+/-) mice produced significantly less IL-1ß and IL-6 compared to wild-type mice and were resistant to LPS-induced endotoxic shock. The loss of Zfp191 may suppress systemic inflammation by reducing these cytokine levels during LPS-induced endotoxic shock.