RESUMEN
AIMS/HYPOTHESIS: The TGF-ß/MAD homologue (SMAD) and nuclear factor κB (NF-κB) signalling pathways have been shown to play a critical role in the development of renal fibrosis and inflammation in diabetic nephropathy. We therefore examined whether targeting these pathways by a kidney-targeting Smad7 gene transfer has therapeutic effects on renal lesions in the db/db mouse model of type 2 diabetes. METHODS: We delivered Smad7 plasmids into the kidney of db/db mice using kidney-targeting, ultrasound-mediated, microbubble-inducible gene transfer. The histopathology, ultrastructural pathology and pathways of TGF-ß/SMAD2/3-mediated fibrosis and NF-κB-dependent inflammation were evaluated. RESULTS: In this mouse model of type 2 diabetes, Smad7 gene therapy significantly inhibited diabetic kidney injury, compared with mice treated with empty vectors. Symptoms inhibited included: (1) proteinuria and renal function impairment; (2) renal fibrosis such as glomerular sclerosis, tubulo-interstitial collagen matrix abundance and renal inflammation, including Inos (also known as Nos2), Il1b and Mcp1 (also known as Ccl2) upregulation, as well as macrophage infiltration; and (3) podocyte and endothelial cell injury as demonstrated by immunohistochemistry and/or electron microscopy. Further study demonstrated that the improvement of type 2 diabetic kidney injury by overexpression of Smad7 was associated with significantly inhibited local activation of the TGF-ß/SMAD and NF-κB signalling pathways in the kidney. CONCLUSIONS/INTERPRETATION: Our results clearly demonstrate that kidney-targeting Smad7 gene transfer may be an effective therapy for type 2 diabetic nephropathy, acting via simultaneous modulation of the TGF-ß/SMAD and NF-κB signalling pathways.
Asunto(s)
Nefropatías Diabéticas/metabolismo , FN-kappa B/metabolismo , Transducción de Señal , Proteínas Smad/metabolismo , Proteína smad7/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Animales , Apoptosis , Complicaciones de la Diabetes/metabolismo , Diabetes Mellitus Tipo 2/sangre , Técnicas de Transferencia de Gen , Inmunohistoquímica/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Microscopía Confocal/métodos , Podocitos/metabolismo , Reacción en Cadena de la Polimerasa/métodos , UltrasonidoRESUMEN
We have convey comparative study of including intensity of marking protein predecessors and nucleic acids by initial strains of Staphylococcus aureus sensitive to antibiotics and strains, which contain plasmids of resistance to different antibiotics. Shown including intensity increase of adenine, uridine, thymidine and marked amino acids by strains which contain plasmids of resistance to different antibiotics to compare with unplasmids variant. On the base of receiving results we may affirm that in general we observe intensification of protein synthesis and separate staves nucleic acid synthesis by observing staunch strains.
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Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Farmacorresistencia Microbiana , Ácidos Nucleicos/metabolismo , Precursores de Proteínas/metabolismo , Staphylococcus aureus/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Staphylococcus aureus/metabolismoRESUMEN
The data submitted suggest that the most effective option in the treatment of rheumatoid arthritis is simultaneous employment of a nonsteroidal anti-inflammatory agent and metatrexate for 1.5 yr and courses of hormonotherapy, exercise therapy, and topical application to the joints of compresses with dimexide, heparin, and analginum as required and ultrasound with hydrocortisone as well. There was no increase in disability rates among patients with rheumatoid arthritis placed on the above combined treatment, which fact is also related to improvement of diagnosis rheumatoid arthritis, well-timed case follow-up, and prescription of courses of prophylactic treatment.
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Artritis Reumatoide/tratamiento farmacológico , Minas de Carbón , Adulto , Antirreumáticos/uso terapéutico , Personas con Discapacidad/estadística & datos numéricos , Femenino , Humanos , Masculino , Persona de Mediana Edad , UcraniaRESUMEN
Intermittent hypoxia treatment (IHT) is gaining attention as a clinical modality due to its capacity to protect cells, tissues, organs, and the whole organism from more intense and/or sustained hypoxia, ischemia and other stresses, to enhance physical and mental capacity. Circulating hematopoietic stem and progenitor cells (HSPC) play an important role in immune response to hypoxia ensuring tissue reparation processes, formation of all types of blood cells etc. There exist considerable individual differences in the capability to mobilize HSPC. This study was designed to compare the effects of IHT on HSPC, various factors of natural resistance and main humoral and cellular components of adaptive immunity in peripheral blood of subjects with normal and reduced tolerance to hypoxic load. Ten healthy male volunteers (age 30,9 +/- 0,6 y.o.) participated in the study and were divided into two groups with reduced (RT, 5 subjects) and normal (NT, 5 subjects) hypoxic tolerance. Criterion for reduced hypoxic tolerance detection was the exceeding deviation of arterial blood pressure, minute ventilation, SaO2 and/or pathological changes in ECG during sustained hypoxia test (breathing with 10% oxygen, 10 min). All subjects were studied before and after a 14 day IHT program consisting of four 5 min bouts/d of breathing 10% O2, with intervening 5 min room air exposures. Immunofluorescence detected HSPCs as CD45+CD34+ cells in peripheral blood. Phagocytic and bactericidal activities of neutrophils, circulating immunoglobulins (IgM, IgG, IgA), immune complexes, complement, and cytokines (EPO, TNF-alpha, IL-4, IFN-gamma) were measured. It was shown that NT subjects had higher hemoglobin and erythrocytes level, hematocrit and physical working capacity, but leukocytes, lymphocytes, CD8+-cells contents and level of IgA were lower than in RT. These differences were preserved after IHT course. CD45+34+cells content was the same in both groups before IHT, but RT subjects demonstrated twice decrease in HSPCs content after IHT as opposed to NT who did not show distinct reactions. A decrease in HSPCs is probably associated with the change of their migration capacity. However, it remains unclear whether there is an inhibition of HSPCs migration into circulation or an activation ofHSPCs escape from circulation. In both cases tissues could accumulate more HSPCs which in turn could enhance hematopoiesis and general regenerative potential. RT group also had lower complement, induced and reserve bactericidal activities of neutrophils which were significantly increased after IHT reaching the level of NT. The level of cytokines EPO, TNF-alpha and IFN-gamma did not differ in both groups before IHT but considerably reduced level of IL-4 was registered in RN patients. IHT sharply lowered pro-inflammatory cytokine TNF-alpha in both groups, significantly increased IL-4 in RT subjects; increasing behavior of IFN-gamma was observed in both groups. EPO was not affected considerably during the study. The findings support the potential for eventual application of IHT for immunotherapy, especially for patients with reduced hypoxic tolerance.
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Células Madre Hematopoyéticas/efectos de los fármacos , Hipoxia/inmunología , Hipoxia/patología , Oxígeno/uso terapéutico , Adaptación Fisiológica/inmunología , Adulto , Complejo Antígeno-Anticuerpo/sangre , Presión Sanguínea , Recuento de Células , Movimiento Celular/efectos de los fármacos , Proteínas del Sistema Complemento/análisis , Citocinas/sangre , Electrocardiografía , Eritropoyetina/sangre , Eritropoyetina/inmunología , Hemoglobinas/análisis , Humanos , Hipoxia/fisiopatología , Inmunidad Celular , Inmunidad Humoral , Masculino , Fagocitosis/efectos de los fármacos , Fagocitosis/inmunología , Intercambio Gaseoso Pulmonar/efectos de los fármacos , Intercambio Gaseoso Pulmonar/inmunología , Respiración/efectos de los fármacosRESUMEN
In the review the information about biological properties of liposomes and their application perspectives as independent medications so as transmitters of medicinal preparations is generalized. Approaches to the directed transport of biologically active matters by the different liposome types are discussed. Simultaneously solving of two important tasks on the decline of preparation toxicity and increase of their efficiency are proposed. The data about liposome distributing features at the different ways of their infusion in an organism and the mechanisms of interaction of corpuscles with the surface of target cells are provided. The most perspective aspects of liposomes and drugs in liposomal form at the modern stage of development of biotechnology, pharmacology and immunopharmacology and their practical application in oncology, antibiotic therapies, chemotherapy of infectious diseases, diabetes, gene engineering, treatment of Parkinson's syndrome, creation of artificial oxygen transporters, immunotherapy, allergology, vaccination and other are discussed.
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Portadores de Fármacos , Liposomas , Preparaciones Farmacéuticas/administración & dosificación , Animales , Portadores de Fármacos/administración & dosificación , Portadores de Fármacos/química , Humanos , Liposomas/administración & dosificación , Liposomas/química , Liposomas/farmacología , Liposomas/uso terapéutico , Preparaciones Farmacéuticas/químicaRESUMEN
OBJECTIVES: Systemic lupus erythematosus is occasionally accompanied with bacterial infection. Lipopolysaccharide (LPS) from bacteria can accelerate and exacerbate lupus nephritis (LN) in animal models, but some mechanisms underlying the LPS-induced acceleration are still unclear. First, it is not known whether LPS can stimulate mesangial cells (MCs) to secrete the pro-inflammatory cytokine, interleukin (IL)-18. Second, it is also unclear whether LPS and/or IL-18 can induce MC apoptosis. Here, we attempted to clarify the cause-and-effect relationships between LPS stimulation, IL-18 production and MC apoptosis to address the above questions. METHODS: LPS was used to induce accelerated LN in LN-prone mice. LPS and IL-18 were also used to treat cultured MCs isolated from the mice. IL-18 expression and MC apoptosis were investigated by in situ hybridization, the TUNEL method, reverse transcription- polymerase chain reaction (RT-PCR), western blotting, DNA electrophoresis and flow cytometry. NFkappaB was detected by immunofluorescent staining. RESULTS: In the LPS-accelerated LN mice, we observed co-existence of IL-18 expression, hyperplasia, apoptosis, and activated apoptotic signal transduction in MCs, as well as marked neutrophil infiltration in the glomerulus, especially around the mesangial region. In cultured MCs, LPS greatly enhanced IL-18 expression, but did not induce apoptosis, while mouse IL-18 did not induce apoptosis or activate apoptotic signal transduction in MCs. CONCLUSIONS: We conclude that LPS can evoke IL-18 production in MCs, but neither LPS nor IL-18 directly induces apoptosis or activates apoptotic signal transduction in the cells. We infer that LPS-induced IL-18 production by MCs could be a mediator by which LPS accelerates and exacerbates LN.
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Interleucina-18/biosíntesis , Lipopolisacáridos/inmunología , Nefritis Lúpica/inmunología , Células Mesangiales/inmunología , Animales , Apoptosis , Autoanticuerpos/metabolismo , Células Cultivadas , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Hiperplasia , Interleucina-18/genética , Nefritis Lúpica/patología , Células Mesangiales/patología , Ratones , Ratones Endogámicos NZB , FN-kappa B/metabolismo , ARN Mensajero/genética , Translocación Genética , Regulación hacia Arriba/inmunologíaRESUMEN
In animal models of IgA nephropathy, the inevitable endogenous immune response to passively administered antigens alone or in complex with specific IgA mask the exact role each might play in pathogenesis. To delineate the role the immune response might play, we have developed a passive model with exclusive IgA-immune complex-mediated nephropathy in B-cell-deficient (BCD) mice. Glomerular IgA immune deposits were induced by administration of purified IgA antiphosphorylcholine and the specific pneumococcal C-polysaccharide (PnC) antigen daily for 2 weeks into BCD and wild-type (WT) mice. In BCD mice IgA+PnC deposits induced severe glomerular injury and renal dysfunction. In contrast, WT mice developed intense glomerular IgG and IgM and C3 co-deposits of the IgA+PnC with significantly less renal injury. Cytofluorometric analysis revealed that PnC induced in BCD, but not in WT, a rapid and dramatic increase in number of activated CD3(+)/CD69(+) T-cell population. The nuclear factor-kappa B (NF-kappaB) transcription factor was activated early and progressively increased in response to glomerular IgA+PnC deposits. These results suggest that nephritogenic IgA+PnC immune deposits induce glomerular and renal dysfunction through activation of the NF-kappaB. This inflammatory pathway is modulated by the endogenous cellular and antibody response to the antigen affecting the course of IgA nephropathy progression.