RESUMEN
Abnormal angiogenesis plays a pathological role in diabetic nephropathy (DN), contributing to glomerular hypertrophy and microalbuminuria. Slit2/Robo1 signaling participates in angiogenesis in some pathological contexts, but whether it is involved in glomerular abnormal angiogenesis of early DN is unclear. The present study evaluated the effects of Slit2/Robo1 signaling pathway on angiogenesis of human renal glomerular endothelial cells (HRGECs) exposed to a diabetic-like environment or recombinant Slit2-N. To remove the effect of Slit2 derived from mesangial cells, human renal mesangial cells (HRMCs) grown in high glucose (HG) medium (33 mM) were transfected with Slit2 siRNA and then the HG-HRMCs-CM with Slit2 depletion was collected after 48 h. HRGECs were cultured in the HG-HRMCs-CM or recombinant Slit2-N for 0, 6, 12, 24, or 48 h. The mRNA and protein expressions of Slit2/Robo1, PI3K/Akt and HIF-1α/VEGF signaling pathways were detected by quantitative real-time PCR, western blotting, and ELISA, respectively. The CCK-8 cell proliferation assay, flow cytometry and the scratch wound-healing assay were used to assess cell proliferation, cycles, and migration, respectively. Matrigel was used to perform a tubule formation assay. Our results showed that the HG-HRMCs-CM with Slit2 depletion enhanced the activation of Slit2/Robo1, PI3K/Akt, and HIF-1α/VEGF signaling in HRGECs in time-dependent manner (0-24 h post-treatment). In addition, the HG-HRMCs-CM with Slit2 depletion significantly promoted HRGECs proliferation, migration, and tube formation. Pretreatment of HRGECs with Robo1 siRNA suppressed the activation of PI3K/Akt and HIF-1α/VEGF signaling and inhibited angiogenesis, whereas PI3K inhibitor suppressed HIF-1α/VEGF signaling, without influencing Robo1 expression. In the HRGECs treated with Slit2-N, Slit2-N time-dependently enhanced the activation of Robo1/PI3K/Akt/VEGF pathway but not HIF-1α activity, and promoted HRGECs proliferation, migration, and tube formation. The effects induced by Slit2 were also abolished by Robo1 siRNA and PI3K inhibitor. Taken together, our findings indicate that in a diabetic-like environment, in addition to mesangial cells, autocrine activation of Slit2/Robo1 signaling of HRGECs may contribute to angiogenesis of HRGECs through PI3K/Akt/VEGF pathway; therefore, Slit2/Robo1 signaling may be a potent therapeutic target for the treatment of abnormal angiogenesis in early DN and may have broad implications for the treatment of other diseases dependent on pathologic angiogenesis.
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Diabetes Mellitus/metabolismo , Células Endoteliales/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Glomérulos Renales/metabolismo , Neovascularización Patológica/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Receptores Inmunológicos/metabolismo , Transducción de Señal , Diabetes Mellitus/genética , Diabetes Mellitus/patología , Células Endoteliales/patología , Humanos , Péptidos y Proteínas de Señalización Intercelular/genética , Glomérulos Renales/patología , Neovascularización Patológica/genética , Neovascularización Patológica/patología , Proteínas del Tejido Nervioso/genética , Receptores Inmunológicos/genética , Proteínas RoundaboutRESUMEN
Cyclophilin D (CypD) is an important component in mitochondrial-dependent tubular cell death in acute kidney injury. However, it is not known whether CypD contributes to tubular cell damage in chronic interstitial fibrosis. We investigated this question in the unilateral ureter obstruction (UUO) model of renal interstitial fibrosis. Groups of CypD-/- and wild type (WT) mice were killed 7 or 12 days after UUO surgery. The significant tubular cell apoptosis seen in WT UUO was significantly reduced in CypD-/- UUO based on TUNEL and cleaved caspase 3 staining. Other markers of tubular cell damage; loss of E-cadherin and AQP1 expression, were also reduced in the CypD-/- UUO kidney. This reduced tubular damage was associated with less inflammation and a partial protection against loss of peritubular capillaries. The prominent accumulation of α-SMA+ myofibroblasts and interstitial collagen deposition seen in WT UUO was significantly reduced in CypD-/- UUO on day 12, but not day 7. Activation of several pro-fibrotic signalling pathways (p38 MAPK, JNK and Smad3) was unaltered in CypD-/- UUO, arguing that CypD acts independently to promote renal fibrosis. CypD deletion in cultured tubular cells attenuated oxidative stress-induced pro-inflammatory, pro-fibrotic and apoptotic responses; however, responses to angiotensin II and LPS were unaffected. In contrast, CypD deletion in cultured renal fibroblasts did not affect PDGF-induced proliferation or TGF-ß1-induced collagen I expression, suggesting no direct role of CypD in the fibroblast response. In conclusion, we have identified a role for CypD in chronic tubular cell damage and in the development of renal interstitial fibrosis.
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Ciclofilinas/metabolismo , Células Epiteliales/metabolismo , Enfermedades Renales/patología , Túbulos Renales/patología , Obstrucción Ureteral/patología , Animales , Células Cultivadas , Peptidil-Prolil Isomerasa F , Ciclofilinas/genética , Fibrosis , Regulación de la Expresión Génica/fisiología , Túbulos Renales/citología , Ratones , Ratones Noqueados , Obstrucción Ureteral/metabolismoRESUMEN
Nephrotoxicity is a common complication of cisplatin chemotherapy and thus limits the use of cisplatin in clinic. The purinergic 2X7 receptor (P2X7R) plays important roles in inflammation and apoptosis in some inflammatory diseases; however, its roles in cisplatin-induced nephrotoxicity remain unclear. In this study, we first assessed the expression of P2X7R in cisplatin-induced nephrotoxicity in C57BL/6 mice, and then we investigated the changes of renal function, histological injury, inflammatory response, and apoptosis in renal tissues after P2X7R blockade in vivo using an antagonist A-438079. Moreover, we measured the changes of nod-like receptor family, pyrin domain containing proteins (NLRP3) inflammasome components, oxidative stress, and proapoptotic genes in renal tissues in cisplatin-induced nephrotoxicity after treatment with A-438079. We found that the expression of P2X7R was significantly upregulated in the renal tubular epithelial cells in cisplatin-induced nephrotoxicity compared with that of the normal control group. Furthermore, pretreatment with A-438079 markedly attenuated the cisplatin-induced renal injury while lightening the histological damage, inflammatory response and apoptosis in renal tissue, and improved the renal function. These effects were associated with the significantly reduced levels of NLRP3 inflammasome components, oxidative stress, p53 and caspase-3 in renal tissues in cisplatin-induced nephrotoxicity. In conclusions, our studies suggest that the upregulated activity of P2X7R might play important roles in the development of cisplatin-induced nephrotoxicity, and P2X7R blockade might become an effective therapeutic strategy for this disease.
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Lesión Renal Aguda/metabolismo , Caspasa 3/metabolismo , Cisplatino/toxicidad , Inflamasomas/metabolismo , Estrés Oxidativo/fisiología , Antagonistas del Receptor Purinérgico P2X/farmacología , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/prevención & control , Animales , Relación Dosis-Respuesta a Droga , Inflamasomas/antagonistas & inhibidores , Masculino , Ratones , Ratones Endogámicos C57BL , Estrés Oxidativo/efectos de los fármacos , Antagonistas del Receptor Purinérgico P2X/uso terapéutico , Piridinas/farmacología , Piridinas/uso terapéutico , Receptores Purinérgicos P2X7/metabolismo , Tetrazoles/farmacología , Tetrazoles/uso terapéuticoRESUMEN
Congenital anomalies of kidney and urinary tract syndrome with or without hearing loss, abnormal ears, or developmental delay (CAKUTHED) is a rare autosomal dominant disorder and variants in PBX1 are involved in the etiology of this syndrome. Precise diagnosis is difficult without genetic test. We described a Chinese CAKUTHED patient, whose characteristics were collected from medical records. The potential responsible variants were explored by whole exome sequencing. A heterozygous variant in the PBX1 gene (NM_002585 c.862C>T, p.R288*) was found in the proband, which was confirmed by Sanger sequencing. This heterozygous variant in the PBX1 gene was the molecular pathogenic basis of this disorder. It is necessary to perform a genetic test for diagnosing chronic nephritis with unknown reason.
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Sordera , Pueblos del Este de Asia , Anomalías Urogenitales , Humanos , Sordera/genética , Secuenciación del Exoma , Pruebas Genéticas , Heterocigoto , Mutación , Linaje , Factor de Transcripción 1 de la Leucemia de Células Pre-B/genética , Anomalías Urogenitales/genéticaRESUMEN
The radiation tolerance of energy storage batteries is a crucial index for universe exploration or nuclear rescue work, but there is no thorough investigation of Li metal batteries. Here, we systematically explore the energy storage behavior of Li metal batteries under gamma rays. Degradation of the performance of Li metal batteries under gamma radiation is linked to the active materials of the cathode, electrolyte, binder, and electrode interface. Specifically, gamma radiation triggers cation mixing in the cathode active material, which results in poor polarization and capacity. Ionization of solvent molecules in the electrolyte promotes decomposition of LiPF6 along with its decomposition, and molecule chain breaking and cross-linking weaken the bonding ability of the binder, causing electrode cracking and reduced active material utilization. Additionally, deterioration of the electrode interface accelerates degradation of the Li metal anode and increases cell polarization, hastening the demise of Li metal batteries even more. This work provides significant theoretical and technical evidence for development of Li batteries in radiation environments.
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Monolithic silica and polymer capillary columns with ultrahigh column efficiencies were prepared. Permeability and electrochromatography performances of these two kinds of columns were compared. Monolithic silica columns bear higher permeability than polymer counterparts by two orders of magnitude. Furthermore, the van Deemter plots of alkylbenzenes on these two kinds of columns demonstrate that monolithic silica columns produce much lower plate heights of alkylbenzes than polymer columns do. Within the range of electroosmotic flow (EOF) velocity investigated, no uptrend of plate height with the increase of EOF was observed suggesting the great capacity of fast separation and high efficiency. The plate height of thiourea on monolithic silica columns is as low as 2.67 µm, representing its corresponding column efficiency is over 430,000 plates/m. As far as we know, it is the highest ever column efficiency reported in the literature. Moreover, the separation of butylbenzene isomers was obtained on the monolithic silica column.
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Polímeros , Dióxido de SilicioRESUMEN
Immunomodulatory enzyme indoleamine 2, 3-dioxygenase (IDO) is one of the initial and rate-limiting enzymes involved in the catabolism of the essential amino acid tryptophan. Via catalysing tryptophan degradation, IDO suppresses adaptive T cell-mediated immunity and plays an important role in various forms of immune tolerance. Its role in T helper type 1 (Th1)-directed, cell-mediated crescentic glomerulonephritis (GN) is still unclear. Therefore, we investigated the activity and role of IDO in crescentic GN using a model of nephrotoxic serum nephritis (NTN), and IDO activity was inhibited by 1-methyl-tryptophan (1-MT) in vivo. Our results showed that activity of IDO, as determined by high performance liquid chromatography analysis of the kynurenine/tryptophan ratio, was increased markedly in the serum and renal tissue of NTN mice, and immunohistochemistry revealed that expression of IDO was up-regulated significantly in glomeruli and renal tubular epithelial cells during NTN. Treatment with 1-MT resulted in significantly exacerbated kidney disease with increased glomerular crescent formation, accumulation of CD4(+)T cells and macrophages in renal tissue, and augmented renal injury compared with phosphate-buffered saline-treated NTN mice, which was associated with enhanced Th1 responses and intrarenal cellular proliferation. These findings suggest that the development of NTN was regulated negatively by increased IDO activity, and IDO might play an important role in the pathogenesis of crescentic GN.
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Glomerulonefritis Membranoproliferativa/enzimología , Indolamina-Pirrol 2,3,-Dioxigenasa/antagonistas & inhibidores , Glomérulos Renales/enzimología , Túbulos Renales/enzimología , Triptófano/análogos & derivados , Animales , Linfocitos T CD4-Positivos/inmunología , Cromatografía Líquida de Alta Presión , Glomerulonefritis Membranoproliferativa/inmunología , Glomerulonefritis Membranoproliferativa/metabolismo , Indolamina-Pirrol 2,3,-Dioxigenasa/análisis , Glomérulos Renales/inmunología , Túbulos Renales/inmunología , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Triptófano/farmacologíaRESUMEN
OBJECTIVE: To study the protective effects of metanephric mesenchymal cells (MMCs) on acute renal tubular damage and explore its possible mechanism. METHODS: MMCs were isolated and cultured from 13-day-old embryonic rats and labeled with 5-bromodeoxyuridine. Seventy-two male SD rats were randomly divided into 3 equal groups: MMC group, receiving MMC injection instantaneously when ischemia/reperfusion (I/R) renal injury was induced, I/R group, undergoing I/R to establish acute renal tubular damage models, and sham operation group. Six rats from each group were killed at different time points: 24 h, 48 h, 72 h, and 96 h later. Blood sample was collected from the vena cava inferior, to examine the serum creatinine (SCr) and blood urea nitrogen (BUN). Specimens of kidney underwent microscopy. Apoptosis was conformed by TUNEL assay. Immunohistochemistry was used to detect the protein expression of Bcl-2 and Bax. The distribution of MMCs labeled with 5-bromodeoxyuridine in kidney was observed by immunofluorescence technique. RESULTS: The SCr and BUN levels in different time points of the MMC group were both significantly lower than those of the I/R group (both P <0.05), HE staining showed that pathological damage of the MMC group was less than that of the I/R group (P <0.05). TUNEL results investigated that the number of apoptosis renal tubular epithelial cells of the MMC group was (13.4 +/- 3.2/HPF), significantly less than that of the I/R group [(25.4 +/- 5.2/HPF)]. In comparison with the I/R group, there were more Bcl-2 positive cells and fewer Bax positive cells in the MMC group. BrdU-labeled MMCs began to occur in the renal tissue (60 +/- 6/HP) In the 72 h subgroup of MMC group, and number of BrdU-labeled MMCs, the 96 h subgroup was (143 +/- 8/HP), significantly higher than that of the 72 h subgroup (P<0.05). CONCLUSION: MMCs have the ability to protect renal function in acute renal tubular damage in rats, migrate and repopulate in the I/R injured renal tubules, and inhibits renal tubular epithelial cell apoptosis. The mechanism may be involved in regulating the expression of Bcl-2 and Bax.
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Apoptosis , Enfermedades Renales/cirugía , Túbulos Renales/cirugía , Mesodermo/trasplante , Enfermedad Aguda , Animales , Nitrógeno de la Urea Sanguínea , Trasplante de Células/métodos , Creatinina/sangre , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Enfermedades Renales/sangre , Enfermedades Renales/etiología , Túbulos Renales/irrigación sanguínea , Túbulos Renales/patología , Masculino , Mesodermo/citología , Mesodermo/metabolismo , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/complicaciones , Proteína X Asociada a bcl-2/metabolismoRESUMEN
OBJECTIVE: To summarize the clinical experience in video-assisted thoracic surgery (VATS). METHODS: From December 1993 to December 2005 1264 patients, 894 males and 370 females, aged 38.9 +/- 12.0, underwent VATS, including bullectomy in 622 cases, resection of mediastinal tumor or cyst in 119 cases, resection of esophageal diseases in 107 cases, lobectomy or wedge-shaped lung resection in 215 cases, lung volume reduction surgery (LVRS) in 17 cases, treatment of thoracic injury in 28 cases, treatment of other thoracic diseases in 72 cases, and biopsy in 84 cases. For the resection of esophageal carcinoma VATS was conducted via the right approach, the esophagus was dissociated, the lymph nodes were resected, upper-abdominal incision was made, the stomach was dissociated and drawn up to the neck region, a cervical incision was made to anastomose the stomach and the residue of esophagus. RESULTS: Operation was completed by VATS successfully in 1230 patients, and 34 cases were converted to traditional thoracotomy because of thoracic adhesion or to radically treat the malignant tumors. Major complications occurred in 45 cases (3.56%), including air-leak lasting more than 7 days in 30 cases, post-operative bleeding in 4 cases (3 of which received VATS once more for hemostasis and the other underwent thoracotomy), hydrothorax or pneumothorax in 3 cases that underwent water-closed drainage, esophageal mucous rupture in 4 cases with achalasia and one case with leiomyoma, all of which underwent repair immediately, infection of pleural cavity in one case after the resection of esophageal diverticulum, and pneumonia in one case after LVRS. One patient with spontaneous pneumothorax and respiratory failure died 5 days after the bullectomy. Spontaneous pneumothorax occurred in 10 patients 2 months to 2 years after VATS 3 of which underwent bullectomy and pleurodesis by VATS once more. CONCLUSION: Spontaneous pneumothorax and some benign thoracic diseases are the major indications of VATS; however, great care should be expended to decide to treat malignant diseases by VATS. It is very important to train the surgeons who are to practice VATS. The practice of VATS should be individualized.
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Enfermedades Torácicas/cirugía , Cirugía Torácica Asistida por Video/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Procedimientos Quirúrgicos Mínimamente Invasivos/efectos adversos , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Complicaciones Posoperatorias/prevención & control , Estudios Retrospectivos , Cirugía Torácica Asistida por Video/efectos adversos , Resultado del TratamientoRESUMEN
OBJECTIVE: To analyze the clinical features and prognosis of patients suffering from fish bile poisoning. METHODS: A total of 156 multiple organ failure (MOF) patients caused by fish bile poisoning were hospitalized in our department over the past 28 years. The patients' symptoms, examination results, treatment and outcomes were collected and analyzed. RESULTS: All patients' first symptom was gastrointestinal discomfort, including unbearable nausea and intractable vomiting. The symptoms that followed were oliguria or anuria, edema, headache, fatigue, jaundice, palpitation, alimentary tract hemorrhage, gross hematuria, dyspnea, shock, tachycardia, bradycardia, arrhythmia, coma, and cardiac arrest. The symptom severity and cohort were different among different patients. Twenty-one cases received gastroscopy, which exhibited diffuse gastric mucosal bleeding. Twelve patients received renal biopsy, which exhibited focal necrosis of tubular epithelial cells. One patient received a liver biopsy, which exhibited extensive hepatocyte necrosis. All patients received blood purification therapy. Of the four patients who died, 4 out of 5 organs had failed. The general mortality rate was 2.6%. CONCLUSIONS: Compared with the MOF caused by trauma and sepsis, the fish bile poisoning MOF has a much lower morality rate. However, patients with higher age, more underlying diseases, and more organ failure tended to have a worse prognosis.
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BACKGROUND: To explore the feasibility of extended resection in selective patients with centrally located lung cancer. METHODS: From January, 1987 to December, 2001, lobectomy or pneumonectomy combined with extended resection of trachea, bronchus, heart or great vessels were carried out in 134 patients with centrally located lung cancer. The operations included bronchoplastic procedures in 80 cases, extended resection and reconstruction of left atrium and/or great vessels in 54 cases (32 cases with contemporary bronchoplasty). RESULTS: Operative death occurred in one case. Postoperative complications happened in 16 cases (11.9%). One hundred and seventeen cases (94.4%) were followed up. The 1-, 3-, 5-year survival rate was 84.7% (61/72), 56.7% (34/60) and 45.7% (21/46) respectively, while of those combined with tracheo bronchoplasty and/or cardiovascular reconstruction, the 1-, 3-, 5-year survival rate was 69.2% (36/52), 46.8% (22/47) and 22.2% (8/36) respectively. (P < 0.05), while expression of KAI1 mRNA did not relate to mutant P53 protein expression (P > 0.05). CONCLUSIONS: Extended resection combined with tracheo-bronchoplasty and/or cardiovascular reconstruction is feasible for selected patients with centrally located lung cancer and could improve the survival and life quality of patients.
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BACKGROUND: Indoleamine 2, 3-dioxygenase (IDO), a heme-containing dioxygenase, can catalyze tryptophan degradation and produce a local microenvironment with tryptophan depletion and tryptophan metabolites accumulation, which may suppress T cell-mediated immunity and play an important immunosuppressive role in many diseases. Previous studies suggested that tryptophan depletion is an important immunosuppressive mechanism of IDO, while recent evidence shows that tryptophan metabolites may also be useful for inducing the T cell immune tolerance. However, it remains unclear whether tryptophan catabolites play a protective role in anti-glomerular basement membrane (anti-GBM) glomerulonephritis (GN), which is a type 1 T-helper (Th1)-mediated autoimmune disease. METHODS: We examined the effect of tryptophan catabolites, 3-hydroxykynurenine acid and 3-hydroxyanthranilic acid, on renal injury in experimental autoimmune glomerulonephritis (EAG) of Wistar-Kyoto rats and explored their protective mechanism. RESULTS: Treatment by either 3-hydroxyanthranilic acid or 3-hydroxykynurenic acid attenuated the kidney disease of EAG rats, with decreased glomerular histological injury and inflammatory cell infiltration, lightened urinary protein, and improved renal function compared to phosphate buffered saline-treated EAG rats. This was associated with significantly increased apoptosis and decreased proliferation of splenic activated T cells in vivo, inducing the deviation of cytokines of antigen-special T cells from Th1 to Th2. CONCLUSIONS: Tryptophan metabolites play an important immunosuppressive role in the development of anti-GBM GN and might offer a new strategy for treating this disease.
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Ácido 3-Hidroxiantranílico/farmacología , Enfermedad por Anticuerpos Antimembrana Basal Glomerular/inmunología , Enfermedad por Anticuerpos Antimembrana Basal Glomerular/patología , Depuradores de Radicales Libres/farmacología , Tolerancia Inmunológica/efectos de los fármacos , Quinurenina/análogos & derivados , Triptófano/metabolismo , Animales , Enfermedad por Anticuerpos Antimembrana Basal Glomerular/enzimología , Apoptosis/efectos de los fármacos , Linfocitos T CD4-Positivos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Tasa de Filtración Glomerular , Indolamina-Pirrol 2,3,-Dioxigenasa/metabolismo , Quinurenina/farmacología , Macrófagos , Proteinuria/etiología , Ratas , Ratas Endogámicas WKY , Células TH1/metabolismo , Células Th2/metabolismoRESUMEN
BACKGROUND: Activation and infiltration of T cells and macrophages are key features of renal tubulointerstitial injury. The costimulatory molecule V-set and immunoglobulin domain-containing protein-4 (VSIG4), which is exclusively expressed on macrophages, is capable of inhibiting the T cell response. However, it is unclear whether VSIG4 is involved in renal tubulointerstitial injury. This study was designed to investigate the role of VSIG4 in renal tubulointerstitial injury and the related T cell infiltration. METHODS: The unilateral ureteric obstruction (UUO) model of renal inflammation and tubulointerstitial fibrosis was established in VSIG4 transgenic knock-out C57BL/6 mice (VSIG4(-/-)) and wild-type C57BL/6 mice (VSIG4(+/+)). Comparative analysis of renal biological indices were assessed by quantitative real-time PCR and immunofluorescence staining. RESULTS: Both the VSIG4(-/-) and VSIG4(+/+) mice showed UUO-related temporal changes in renal expression of CD3, CD4 and CD8 T cell markers, with the protein levels being significantly lower in the VSIG4(+/+) UUO mice. Moreover, at each time point examined the UUO VSIG4(+/+) mice showed significantly lower renal mRNA levels of the cytokines interleukin (IL)-2, interferon- and tumor necrosis factor-, but significantly higher IL-10, than the UUO VSIG4(-/-) mice. CONCLUSIONS: The macrophage-expressed VSIG4 may act to alleviate renal tubulointerstitial injury via inhibition of T cell infiltration and secretion of inflammation related factors.
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Túbulos Renales/patología , Macrófagos/metabolismo , Nefritis/inmunología , Nefritis/patología , ARN Mensajero/metabolismo , Receptores de Complemento/metabolismo , Animales , Complejo CD3/metabolismo , Antígenos CD4/metabolismo , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos , Antígenos CD8/metabolismo , Linfocitos T CD8-positivos , Fibrosis , Interferón gamma/genética , Interleucina-2/genética , Interleucinas/genética , Activación de Linfocitos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores de Complemento/genética , Factor de Necrosis Tumoral alfa/genética , Obstrucción Ureteral/complicacionesRESUMEN
Neutral endopeptidase (NEP) is the first target antigen identified on podocytes in human membranous nephropathy (MN). Cytotoxic T lymphocytes (CTLs) are considered essential for glomerular destruction in MN model. The aim of this study was to show that the CTL epitopes of NEP could be used to design more effective and better tolerated therapies. The CTL epitopes of NEP were screened using the long-distance prediction system SYFPEITHI and the Bioinformatics and Molecular Analysis Section of the MHC Peptide Binding Predictions program. Peptides were synthesized and immunoreactivity was assessed by peptide-MHC-binding affinity assay, cytotoxicity assay and HLA-A2.1/Kb transgenic mice immunization. Five candidates were identified according to the high scores generated by the computer predicting system. Peptide NEP(375-383) (FIMDLVSSL), which up-regulated HLA-A2.1 molecular expression, showed a high affinity to HLA-A2.1, whereas NEP(268-276), NEP(297--305) and NEP(492-500) (QLALEMNKV, MLLYNKMRL and KLNNEYLEL) showed a moderate affinity and NEP(559-567) (ILQPPFFSA) only had a low affinity. Cytotoxicity assay further showed that NEP(268-276) and NEP(375-383) could induce NEP-specific CTL responses in vitro. Unexpectedly, we found that a single CTL epitope, NEP(375-383), could induce proteinuria and glomerular injury in HLA-A2.1/K(b) transgenic mice in vivo. HLA-A*0201-restricted CTL epitope NEP(375-383) can serve as a potential candidate for designing MN vaccine.
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Linfocitos T CD8-positivos/inmunología , Epítopos de Linfocito T/inmunología , Glomerulonefritis Membranosa/inmunología , Antígeno HLA-A2/inmunología , Neprilisina/inmunología , Podocitos/inmunología , Animales , Linfocitos T CD8-positivos/patología , Simulación por Computador , Mapeo Epitopo/métodos , Epítopos de Linfocito T/genética , Femenino , Glomerulonefritis Membranosa/enzimología , Glomerulonefritis Membranosa/genética , Glomerulonefritis Membranosa/patología , Glomerulonefritis Membranosa/terapia , Antígeno HLA-A2/genética , Humanos , Masculino , Ratones , Ratones Transgénicos , Neprilisina/genética , Podocitos/enzimología , Podocitos/patología , Proteinuria/enzimología , Proteinuria/genética , Proteinuria/inmunología , Proteinuria/patología , Proteinuria/terapiaRESUMEN
Sinomenine has been used to treat autoimmune diseases for centuries. However, the mechanism underlying its therapeutic effects remains unknown. Increasing recognition of the importance of the Th1/Th2 imbalance in nephritis has raised the questions of whether there is a Th1/Th2 imbalance in patients with mesangial proliferative nephritis (MsPGN) and whether sinomenine can modulate the Th1/Th2 imbalance. In this study, 25 MsPGN patients were treated with sinomenine and followed for 3 months. The expression of T-bet and GATA-3 mRNA in peripheral blood mononuclear cells (PBMCs) and the serum levels of interferon-gamma (IFN-gamma), interleukin (IL)-4, and IL-10 were studied at month 0, month 1, and month 3. The intra-renal expression of T-bet and GATA-3 was studied via immunohistochemistry. Results reveal that PBMCs from MsPGN patients expressed high levels of T-bet mRNA and low levels of GATA-3 mRNA, and the T-bet/GATA-3 ratio in MsPGN patients was significantly higher than that in healthy donors. Meanwhile, MsPGN patients were found to have simultaneously elevated IFN-gamma values and decreased IL-10 values. Immunohistochemistry revealed increased T-bet and decreased GATA-3 expression in renal tissues from MsPGN patients. Moreover, sinomenine was found to cause a decrease in T-bet mRNA expression, resulting in a drop in the T-bet/GATA-3 ratio. Sinomenine was also found to elicit a decrease in the serum levels of IFN-gamma. These results suggest that a shift toward the Th1 pathway of Th cell activation occurs in MsPGN patients, and that sinomenine has the potential to counter this shift in the Th1/Th2 balance and thereby produce therapeutic effects.
Asunto(s)
Glomerulonefritis Membranoproliferativa/tratamiento farmacológico , Riñón/metabolismo , Morfinanos/administración & dosificación , Células TH1/metabolismo , Células Th2/metabolismo , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios de Seguimiento , Factor de Transcripción GATA3/genética , Factor de Transcripción GATA3/inmunología , Factor de Transcripción GATA3/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Glomerulonefritis Membranoproliferativa/sangre , Glomerulonefritis Membranoproliferativa/genética , Glomerulonefritis Membranoproliferativa/patología , Homeostasis , Humanos , Inmunohistoquímica , Interferón gamma/sangre , Interleucina-10/sangre , Interleucina-4/sangre , Riñón/efectos de los fármacos , Riñón/patología , Masculino , Persona de Mediana Edad , Proteínas de Dominio T Box/genética , Proteínas de Dominio T Box/inmunología , Proteínas de Dominio T Box/metabolismo , Células TH1/efectos de los fármacos , Células TH1/inmunología , Células TH1/patología , Células Th2/efectos de los fármacos , Células Th2/inmunología , Células Th2/patologíaRESUMEN
OBJECTIVE: To evaluate the feasibility of reconstruction of urothelium tissue in vivo using tissue-engineering technique. METHODS: The urothelium cells were obtained from young rabbit, bladder by mechanical and enzyme digested methods. After expanded in vitro, the 4th to 5th generation urothelium cells were seeded onto the surface of 8 Polylatical/glycolic acid copolymer polymer, the polymer matrix without seeding cells served as control group. A total of 8 cell-polymer scaffolds and 4 simply scaffolds were separately implanted into subcutaneous pockets of athymic mice. The experiment groups included cell-polymer scaffolds 4 weeks and cell-polymer scaffolds 8 weeks. The control group included simply scaffold 4 weeks and simply scaffold 8 weeks. After 4 and 8 weeks, the specimens were obtained and examined by gross inspection, histologically and immunohistochemically. RESULTS: The results of HE and Masson staining showed that the polymer were covered by urothelium cells layers and cells layers increased markley in experimental group. Immunocytochemical studies revealed that the cells were stained positively for anti-cytokeratins (AE1/AE3) in experimental group. Fiber tissue deposition were found on the surface of polymers in control group by HE and Masson staining. Immunocytochemical staining of implants showed the negative result for cytokeratins in control group. CONCLUSION: It is feasibility that reconstruction of urothelium tissue using tissue-engineering technique,which provides basic understandings for further development of the bladder and ureteral tissue engineered research.