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1.
Blood Purif ; 48(1): 76-85, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31030200

RESUMO

BACKGROUND: Despite the progression of dialysis techniques,  the mortality of hemodialysis (HD) patients is still high in China. Here, a retrospective study was performed to investigate the neglected risk factors of all-cause mortality during maintenance HD (MHD). METHODS: We investigated 117 MHD patients who died between 2011 and 2016 in the Second Xiangya Hospital of Central South University HD center. In order to analyze the risk factors of 48 months all-cause death, the methods of Kaplan-Meier and Cox regression were used. RESULTS: Multivariate analyses of adjusted age and gender showed that MHD patients with estimated glomerular filtration rate <7 or >10 mL/min/1.73 m2 and anemia (hemoglobin <100 g/L) at the initiation of dialysis are independently associated with the higher death risk. Using central venous catheter vascular access, cerebrovascular comorbidities, diabetes, low-flux dialyzer, and dialysis frequency ≤2 times weekly were also the independent risk factors of death within 48 months. CONCLUSIONS: This study indicated that the status of HD initiation is a risk factor of long-term survival in MHD patients, which were usually ignored for lacking of nephrology care prior and could potentially be identified and modified to improve the survival prognosis. Video Journal Club "Cappuccino with Claudio Ronco" at  https://www.karger.com/Journal/ArticleNews/223997?sponsor=52.


Assuntos
Causas de Morte , Falência Renal Crônica/mortalidade , Falência Renal Crônica/terapia , Diálise Renal , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Feminino , Humanos , Estimativa de Kaplan-Meier , Falência Renal Crônica/diagnóstico , Falência Renal Crônica/epidemiologia , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Diálise Renal/efeitos adversos , Diálise Renal/métodos , Análise de Sobrevida , Adulto Jovem
2.
Int Urol Nephrol ; 48(11): 1855-1861, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27465795

RESUMO

OBJECTIVE: The proteins BAFF, ST6GALNAC2, C1GALT1, and COSMC in peripheral blood mononuclear cells (PBMCs) and plasma levels of IgA1 and galactose-deficient IgA1 (Gd-IgA1) are potential biomarkers for IgAN nephropathy. In this study, we comparatively studied the changes of those biomarkers before and after tonsillectomy. METHODS: Peripheral blood samples were obtained from 16 IgAN patients with pre- and post-tonsillectomy. IgAN was diagnosed based on results from analysis of percutaneous renal biopsy tissue. Peripheral blood samples from three patients without renal diseases (non-IgAN), before and after tonsillectomy, and 16 healthy controls were also examined. BAFF, ST6GALNAC2, C1GALT1, and COSMC mRNA levels in PBMCs were detected using real-time PCR. Plasma IgA1 content was measured by ELISA. Gd-IgA1 levels were determined using the VV lectin-ELISA method. RESULTS: BAFF, ST6GALNAC2, C1GALT1, and COSMC mRNA levels and the plasma concentrations of IgA1 and Gd-IgA1 in IgAN patients before tonsillectomy were significantly higher than those in healthy controls (P < 0.05). Tonsillectomy significantly increased the expression of BAFF and ST6GALNAC2, and plasma IgA1 level, while it downregulated that of C1GALT1 and COSMC (P < 0.05). However, in non-IgAN patients, tonsillectomy did not affect the mRNA levels of BAFF, ST6GALNAC2, C1GALT1, and COSMC, plasma IgA1 content and Gd-IgA1 level. Positive correlations were established between BAFF and IgA1 (r = 0.604, P < 0.01) and between ST6GALNAC2 and Gd-IgA1 (r = 0.623, P < 0.01). CONCLUSIONS: Tonsillectomy changes the mRNA levels of BAFF, ST6GALNAC2, C1GALT1, and COSMC in PBMCs, as well as the plasma IgA1 level in IgAN patients. BAFF and ST6GALNAC2 might regulate IgA1 secretion and O-glycosylation.


Assuntos
Glomerulonefrite por IGA/sangue , Glomerulonefrite por IGA/diagnóstico , Imunoglobulina A/sangue , RNA Mensageiro/sangue , Tonsilectomia , Adulto , Fator Ativador de Células B/genética , Biomarcadores/sangue , Estudos de Casos e Controles , Feminino , Galactosiltransferases/genética , Expressão Gênica , Glomerulonefrite por IGA/genética , Humanos , Leucócitos Mononucleares , Masculino , Chaperonas Moleculares/genética , Período Pós-Operatório , Período Pré-Operatório , Estudos Prospectivos , Sialiltransferases/genética , Adulto Jovem
3.
Beijing Da Xue Xue Bao Yi Xue Ban ; 47(5): 749-53, 2015 Oct 18.
Artigo em Chinês | MEDLINE | ID: mdl-26474610

RESUMO

OBJECTIVE: To observe an abnormal expression of humoral immune response induced by memory B cells in tonsils and peripheral blood of patients with IgA nephropathy (IgAN), the variation of memory B cells after tonsillectomy, and to discover the role of tonsillectomy in IgAN. METHODS: In the study, 28 patients were diagnosed as IgAN via renal biopsy, and 27 patients suffering from chronic tonsillitis without nephritis and 10 normal human beings were selected as controls. The expression of memory B cells in the tonsils and peripheral blood was tested by flow cytometry, and the same method was used to test the variation of the expression of memory B cells in peripheral blood of patients with IgAN after tonsillectomy. RESULTS: In this study, higher percentages of memory B cells were observed in tonsil and peripheral blood of IgAN patients, which were 5.72%±5.26%, 4.92%±5.10%. After tonsillectomy, the percentage of memory B cells was 1.10%±0.65%, lower than that before tonsillectomy (P<0.05). Meanwhile, in tonsils and peripheral blood, the percentage of memory B cells varied with the variation of the urinary findings of the IgAN patients. CONCLUSION: The percentage of memory B cell in tonsils and peripheral blood could predict disease progression of IgAN to a certain extent.


Assuntos
Subpopulações de Linfócitos B/imunologia , Glomerulonefrite por IGA/fisiopatologia , Tonsila Palatina/citologia , Estudos de Casos e Controles , Doença Crônica , Progressão da Doença , Citometria de Fluxo , Humanos , Tonsila Palatina/imunologia , Tonsilectomia , Tonsilite
4.
Am J Transl Res ; 7(11): 2199-211, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26807168

RESUMO

Norcantharidin (NCTD) has been proven to be able to attenuate renal interstitial fibrosis, but the exact molecular mechanism is still unknown. This study investigated the relationship between the anti-fibrotic effect of NCTD and its inhibition on PP2Ac expression. Here, PP2Ac was found to be positively correlated with extracellular matrix accumulation in the rat unilateral ureteral obstruction (UUO) model. Additional experiments showed that the PP2A inhibitor (okadaic acid) can ameliorate renal interstitial fibrosis by inhibiting the expression of fibronectin (FN) and collagen I (Col-I) and reversing tubular epithelial-to-mesenchymal transition in vivo and in vitro. In vitro experiments also demonstrated that ectopic over-expression of PP2Ac has a profibrotic effect in HK-2 cells. Moreover, NCTD was able to downregulate PP2Ac expression, decrease FN, Col-I, α-SMA expression, and increase E-cadherin expression in a dose-dependent manner both in vivo and in vitro. In particular, it was demonstrated that NCTD induced no evident changes in the expression of FN, Col-I, α-SMA and E-cadherin in HK-2 cells after PP2Ac was knocked down by shRNA. These results indicated that NCTD exerts an anti-fibrosis effect via inhibition of PP2Ac expression. Thus, PP2Ac could be a promising target for intervention in renal interstitial fibrosis.

5.
Mol Med Rep ; 9(6): 2293-300, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24718692

RESUMO

The present study investigated the potential effects of the long-term expression of exogenous adiponectin (ADPN) on normal and diabetic kidneys. Type 2 diabetes mellitus models were induced by high-lipid and high-sucrose feeding plus intraperitoneal injection of streptozotocin. The recombinant plasmid pIRES2-EGFP-gAd, which is able to co-express globular ADPN (gAd) and enhanced green fluorescent protein (EGFP), was intraperitoneally injected into rat models mediated by Lipofectamine. In total, 32 Wistar rats were randomly assigned into four groups: the normal control group, the diabetes group, the diabetes group treated with pIRES2-EGFP-gAd and the diabetes group treated with pIRES2-EGFP. After 12 weeks, serum biochemistry and urine albumin levels were measured. The kidneys were collected to assess the generation of reactive oxygen species (ROS) and the renal pathological changes were observed by light microcopy. The protein expression of endothelial nitric oxide synthase (eNOS), transforming growth factor-ß1 (TGF-ß1) and phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK) were determined by an immunohistochemical staining method and western blot analysis. Intraperitoneal injection of the human gAd gene via Lipofectamine resulted in abundant ADPN protein in the kidney. In the diabetic rats, the delivery of the exogenous gAd gene ameliorated the progression of diabetic nephropathy (DN). ADPN attenuated urine albumin excretion in the diabetic rats. ADPN also mitigated glomerular mesangial expansion, reduced the generation of ROS and prevented interstitial fibrosis. In addition, the expression of gAd inhibited the renal expression of TGF-ß1, promoted the protein expression of eNOS and activated the opening of the AMPK signaling pathway in the renal tissues of the diabetic rats. Despite the effects of ADPN on DN being controversial, these observations indicate that the supplementation of ADPN is beneficial in ameliorating DN in rats.


Assuntos
Adiponectina/genética , Nefropatias Diabéticas/genética , Expressão Gênica , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Nefropatias Diabéticas/terapia , Modelos Animais de Doenças , Genes Reporter , Mesângio Glomerular/metabolismo , Mesângio Glomerular/patologia , Injeções Intraperitoneais , Rim/metabolismo , Rim/patologia , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Masculino , Óxido Nítrico Sintase Tipo III/metabolismo , Proteinúria/genética , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Transfecção , Fator de Crescimento Transformador beta1/sangue , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/urina
6.
Int J Clin Exp Med ; 7(12): 5517-22, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25664064

RESUMO

AIM: A rapid protocol is necessary to determine the serum concentrations of prednisone. METHODS: The HP1100 high-performance liquid chromatographic (HPLC) system was employed. The HP Lichrosphere C8 column (250 mm × 4 mm, i.d., 5 µm particle size) was used. The mobile phase was methanol, tetrahydrofuran and water in the ratio 25:25:50. The flow rate was 1.0 ml/min. The sample was monitored by UV absorbance at 240 nm. Acetanilide was used as the internal standard, and methanol was added into the serum for depositing the protein. RESULTS: The chromatography was effective and was not interfered with by the serum components. Good linearity was observed, within the range of 10-500 µg/L for prednisone, and the detection limit was 5 µg/L. The serum concentrations of prednisone between the nephrotic syndrome (NS) group and the control group were significantly different (P < 0.05), while there was no significant difference between the females and males of the NS group (P > 0.05). The serum ncentration of prednisone in the steroid-resistant group was lower than that in the steroid-sensitive group (P < 0.05). CONCLUSIONS: HPLC is a practical and reliable method to determine the serum concentration of prednisone with high accuracy, precision, linearity and repeatability.

7.
Ren Fail ; 36(2): 292-9, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24152223

RESUMO

OBJECTIVE: To determine functional and structural alterations of peritoneum and fibrotic cytokines expression in peritoneal dialysis (PD) rats. METHODS: 28 Sprague-Dawley (S-D) rats were randomly divided into four groups and dialyzed with various solutions daily for four weeks: (1) no solution (CON group), (2) 0.9% Saline solution (NS group), (3) 1.5% Dianeal (LG group), (4) 4.25% Dianeal (HG group). Peritoneal equilibration tests, ultrafiltration function and effluent protein quantification were measured. Peritoneum morphology was studied and immunohistochemistry were performed for detection of transforming growth factor ß1 (TGF-ß1), connective tissue growth factor (CTGF), and fibronectin (FN) proteins. Reverse transcriptional-polymerase chain reaction was used to analyze the expression of TGF-ß1, CTGF mRNA. RESULTS: Administration of 4.25% Dianeal caused functional and structural changes of peritoneum, including protein loss through the transport process, decrease of peritoneal solute transport rate and ultrafiltration capacity. The collagen of peritoneum in the HG group was thicker than the other groups. The levels of CTGF, TGF-ß1, and FN proteins were significantly the highest in the HG group, followed by the LG group. The liner correlation analysis showed positive correlations between the levels of CTGF, TGF-ß1, and FN proteins and the collagen thickness. The expression of TGF-ß1 and CTGF mRNA in the HG group were significantly higher than those in the other groups and were indicated positive correlation. CONCLUSION: Using high glucose peritoneal dialysis solutions in rats may not only lead to processing of peritoneal fibrosis, which is promoted by ectopic expression of TGF-ß1, but also increase the expression of CTGF. CTGF is an important fibrotic media of peritoneal fibrosis in PD rats.


Assuntos
Fator de Crescimento do Tecido Conjuntivo/metabolismo , Soluções para Diálise , Glucose/administração & dosagem , Diálise Peritoneal , Peritônio/metabolismo , Peritônio/patologia , Fator de Crescimento Transformador beta1/metabolismo , Animais , Citocinas/metabolismo , Fibronectinas/metabolismo , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Regulação para Cima
8.
Hum Immunol ; 74(6): 708-12, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23313256

RESUMO

BACKGROUND: B cells in tonsil, which may produce the nephritogenic IgA, have been incriminated in the pathogenesis of IgAN. The aim of the present study was to assess the role of memory B cell in clinical progression of IgAN. Methods we investigated 28 IgAN patients and 27 age-matched patients with chronic tonsillitis without IgAN, who were treated by tonsillectomy, meanwhile, the peripheral blood (PB) of 10 healthy individuals were also as control groups. In tonsil and PB, the frequency of memory B cells were tested by Flow cytometric (FCM). RESULTS: In this study, higher percentage of memory B cells were observed in tonsil and PB of IgAN patients. After tonsillectomy, the percentage of memory B cells in IgAN patients were significantly (P<0.05) lower than before tonsillectomy. Meanwhile, in tonsil and PB, the percentage of memory B cells variated with the variation of urinary finding of IgAN patients. CONCLUSIONS: The percentage of memory B cell in tonsil and PB could predict disease progression of IgAN to a certain extend; it's variation in pre- and post- tonsillectomy can provide theoretical basis to cure IgAN patients indirectly.


Assuntos
Linfócitos B/imunologia , Glomerulonefrite por IGA/imunologia , Memória Imunológica , Adulto , Progressão da Doença , Feminino , Glomerulonefrite por IGA/sangue , Hematúria/sangue , Hematúria/imunologia , Humanos , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Tonsila Palatina/imunologia , Fenótipo , Proteinúria/sangue , Proteinúria/imunologia , Tonsilectomia , Adulto Jovem
9.
J Biomed Biotechnol ; 2012: 673096, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23118514

RESUMO

Background. microRNA (miRNA, miR) are thought to interact with multiple mRNAs which are involved in the EMT process. But the role of miRNAs in peritoneal fibrosis has remained unknown. Objective. To determine if miRNA589 regulates the EMT induced by TGFß1 in human peritoneal mesothelial cell line (HMrSV5 cells). Methods. 1. Level of miR589 was detected in both human peritoneal mesothelial cells (HPMCs) isolated from continuous ambulatory peritoneal dialysis (CAPD) patients' effluent and HMrSV5 cells treated with or without TGFß1. 2. HMrSV5 cells were divided into three groups: control group, TGFß1 group, and pre-miR-589+TGFß1 group. The level of miRNA589 was determined by realtime PCR. The expressions of ZO-1, vimentin, and E-cadherin in HPMCs were detected, respectively. Results. Decreased level of miRNA589 was obtained in either HPMCs of long-term CAPD patients or HMrSV5 cells treated with TGFß1. In vitro, TGFß1 led to upregulation of vimentin and downregulation of ZO-1 as well as E-cadherin in HMrSV5 cells, which suggested EMT, was induced. The changes were accompanied with notably decreased level of miRNA589 in HMrSV5 cells treated with TGFß1. Overexpression of miRNA589 by transfection with pre-miRNA589 partially reversed these EMT changes. Conclusion. miRNA589 mediates TGFß1 induced EMT in human peritoneal mesothelial cells.


Assuntos
Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal/genética , MicroRNAs/metabolismo , Peritônio/citologia , Caderinas/metabolismo , Separação Celular , Células Epiteliais/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , MicroRNAs/genética , Diálise Peritoneal Ambulatorial Contínua , Fator de Crescimento Transformador beta1/farmacologia , Vimentina/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
10.
Asian Pac J Trop Med ; 5(7): 561-5, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22647820

RESUMO

OBJECTIVE: To investigate the efficiency of ß-galactosidase gene transfer into rat kidney with ultrasound-mediated microbubble destruction via different injection routes. METHODS: A total of 25 Wistar rats were randomly divided into 5 groups. Four groups received a mixture of optison microbubbles (0.2 mL) and lacz plasmids (25 µg) injection via renal artery, tail vein, anterior tibial muscle and renal parenchyma, respectively. The control group received a mixture of PBS (xx mL) and lacz plasmids (25 µg) via renal artery. Three days after the gene transfer, ultrasound with fixed frequency and power (1 MHz, xxW) was delivered to the kidneys for 3 min. The efficiency of the gene transfer and expression was evaluated on the basis of ß-galactosidase expression. The side effects of this method were evaluated by immunohistological method. RESULTS: ß-galactosidase expression could be observed only in tubules but not in glomeruli and interstitial area. The efficiency of renal artery group was higher than that of tail vein, anterior tibial muscle and renal parenchyma group (P<0.05). Immunohistochemical analysis revealed co-expression of ß-galactosidase with a roximal tubule marker, megalin, which suggested that ultrasound enhanced gene transfer into the proximal tubular epithelial cells. No ß-galactosidase expression was observed in the extrarenal organs. There were no evident pathological and biochemical changes after gene transfer. CONCLUSIONS: Ultrasound-mediated microbubble destruction can transfer gene into kidney via renal artery, tail vein, anterior tibial muscle and renal parenchyma. Compared with renal artery, administrating microbubbles via tail vein and anterior tibial muscle are more convenient and less vulnerarious.


Assuntos
Albuminas/metabolismo , Fluorcarbonetos/metabolismo , Técnicas de Transferência de Genes , Rim/metabolismo , Ultrassom , beta-Galactosidase/genética , Albuminas/administração & dosagem , Animais , Fluorcarbonetos/administração & dosagem , Imuno-Histoquímica , Injeções , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Masculino , Microbolhas , Plasmídeos/administração & dosagem , Plasmídeos/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , beta-Galactosidase/metabolismo
11.
Ren Fail ; 34(7): 821-6, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22679920

RESUMO

OBJECTIVE: To explore the association of urinary podocyte excretion and renal expression of podocyte-specific marker podocalyxin (PCX) with clinicopathological changes in immunoglobulin A nephropathy (IgAN). METHODS: Morning urine samples from IgAN patients and healthy controls were collected. The expression of glomerular PCX was quantified in 50 IgAN patients diagnosed by renal biopsy. IgAN was classified based on the Lee's Grading system and scored according to the Katafuchi semiquantitative criteria. Morphological evaluation of podocyte was determined by electron microscopy. RESULTS: The amount of urinary podocytes in the IgAN patients was significantly higher than that in the healthy controls (p < 0.01). Pairwise comparison among Lee's grades of IgAN showed that the median of urinary podocytes in Lee's I-II group was lower than that in Lee's III, IV, and V groups (p < 0.05); group III lower than group V (p < 0.05). The positive rate of urinary podocytes was the highest in Lee's IV and V groups (100%), and lowest in Lee's I-II group (55%). Multiple comparison among groups of Lee's grades of IgAN showed that the glomerular PCX expression in Lee's I-II group was higher than that in Lee's III, IV, and V groups (p < 0.05); groups III and IV higher than group V (p < 0.05). The amount of urinary podocytes in IgAN patients was negatively correlated with PCX expression (r = -0.702, p < 0.01), but positively correlated with 24-h urinary protein (r = 0.465, p < 0.01) and glomerular (r = 0.233, p < 0.01) and renal tubular pathological scores (r = 0.307, p < 0.05). The glomerular PCX expression was negatively correlated with 24-h urinary protein (r = -0.367, p < 0.05) and glomerular (r = -0.560, p < 0.05) and tubular pathological scores (r = -0.377, p < 0.05). Electron microscopy showed significant changes in podocytes of IgAN, especially in the foot process. CONCLUSION: The amount of urinary podocyte can reflect the loss of podocytes in renal tissue, which may be a marker of IgAN progression.


Assuntos
Glomerulonefrite por IGA/urina , Rim/patologia , Podócitos/citologia , Sialoglicoproteínas/urina , Adolescente , Adulto , Estudos de Casos e Controles , Contagem de Células , Feminino , Glomerulonefrite por IGA/patologia , Humanos , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Podócitos/ultraestrutura , Urina/citologia , Adulto Jovem
12.
Mol Cell Biochem ; 361(1-2): 79-83, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21956672

RESUMO

Progressive renal tubulointerstitial fibrosis is a common final pathway of nearly all forms of chronic kidney disease. Many efforts have been done to arrest or prevent renal tubulointerstitial fibrosis but with little progress. Nowadays, few therapeutic agents are available in clinical use. Norcantharidin (NCTD) is of great benefit in anticancer treatment, by inducing cell apoptosis, inhibiting cell proliferation, in addition, blocking tumor metastasis and angiogenesis in cancer, whereas little attention is given to its relationship with other diseases. Our recent studies demonstrated that NCTD was protective against renal tubulointerstitial fibrosis both in vivo and in vitro. The underlying mechanisms may include modulation of TGF-ß1/Smad signal cascade, inhibition of protein serine/threonine phosphatases (PPP) as well as NF-κB. NCTD may be a promising therapeutic agent for renal tubulointerstitial fibrosis. In the present article, we will review the action of NCTD in renal tubulointerstitial fibrosis and discuss its possible mechanisms.


Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/uso terapêutico , Nefropatias/tratamento farmacológico , Túbulos Renais/patologia , Animais , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Fibrose , Humanos , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Fosfoproteínas Fosfatases/antagonistas & inibidores , Fosfoproteínas Fosfatases/metabolismo , Transdução de Sinais/efeitos dos fármacos
13.
J Nephrol ; 25(4): 481-9, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-21786225

RESUMO

BACKGROUND: There is rapidly accumulating evidence that use of oxidized hair dye causes various forms of nephrotoxic injury. However, the regulation and implication of the nephrotoxic injury resulting from p-aminophenol (PAP) and p-paraphenylenediamine (PPD), the main components of oxidized hair dye, remain unknown. METHODS: To clarify the effect of PAP and PPD, we analyzed the proliferation, lactate dehydrogenase (LDH) levels, apoptosis and subsequent mRNA levels of caspase-3 in HK-2 cells stimulated with different concentrations of PAP or PPD. Cell proliferation was assessed by MTT assay. The production of LDH was determined by Hitachi 7170 biochemical analyzer. The apoptosis of cell was analyzed using flow cytometry and scanning electron microscopy. The mRNA levels of caspase-3 were quantitatively measured by real-time PCR. RESULTS: The proliferation of HK-2 cells was significantly inhibited by PAP, PPD and each mixed with hydrogen peroxide (H2O2). The level of apoptosis of HK-2 cells, the mRNA levels of caspase-3 and LDH production by PAP or PPD stimulation was significantly higher than controls or after H2O2. A typical apoptotic morphological change was observed under electron microscopy in response to PAP or PPD. CONCLUSION: It appears that caspase-3 may play a key role in the nephrotoxic injury resulting from PAP, PPD or its oxidized form.


Assuntos
Aminofenóis/toxicidade , Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Tinturas para Cabelo/toxicidade , Túbulos Renais Proximais/efeitos dos fármacos , Fenilenodiaminas/toxicidade , Caspase 3/genética , Caspase 3/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Epiteliais/metabolismo , Células Epiteliais/ultraestrutura , Citometria de Fluxo , Humanos , Peróxido de Hidrogênio/toxicidade , Túbulos Renais Proximais/metabolismo , Túbulos Renais Proximais/ultraestrutura , L-Lactato Desidrogenase/metabolismo , Microscopia Eletrônica de Varredura , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Medição de Risco , Fatores de Tempo
14.
Lab Invest ; 91(12): 1706-16, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21931301

RESUMO

Norcantharidin (NCTD) was shown in our previous studies to attenuate renal tubulointerstitial fibrosis in rat models with diabetic nephropathy (DN). The aim of this study was to determine the effects of NCTD on the expression of extracellular matrix (ECM) and TGF-ß1 in HK-2 cells stimulated by high glucose and on calcineurin (CaN)/NFAT pathway. Whether or not the antifibrotic effect of NCTD on renal interstitium was dependent on its inhibition of CaN pathway was also investigated. Experimental concentrations of NCTD were verified by cytotoxic test and MTT assay. HK-2 cells were transfected with CaN small interference RNA (siRNA). The mRNA and protein expressions of FN, ColIV, TGF-ß1, and CaN in HK-2 cells were detected by real-time PCR and western blot. The CaN/NFAT pathway was examined by indirect immunofluorescence and western blot. Our study revealed that NCTD concentrations over 5 mg/l had overt cytotoxicity on HK-2 cells. Meanwhile, both 2.5 and 5 mg/l NCTD inhibited HK-2 cell proliferation (P < 0.05). NCTD inhibited the upregulation of FN, ColIV, and TGF-ß1 of HK-2 cells stimulated by high glucose (P < 0.05), and also significantly downregulated the expression of CaN mRNA and protein in HK-2 cells (P < 0.05). In addition, not only was the nuclear translocation of NFATc inhibited, but its protein level in the nucleus was also reduced. Following CaN siRNA transfection, CaN mRNA and protein expression were significantly decreased. In contrast, the protein levels of FN, ColIV, and TGF-ß1 increased in HK-2 cells stimulated by high glucose (P < 0.05). However, NCTD treatment downregulated their expression. These results indicated that NCTD could decrease the expression of ECM and TGF-ß1 in HK-2 cells stimulated by high glucose, downregulate CaN expression, and block the CaN/NFAT signaling pathway. However, the effect of NCTD on inhibition of the expression of ECM and TGF-ß1 was not associated with its inhibition of the CaN/NFAT pathway.


Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/uso terapêutico , Calcineurina/metabolismo , Nefropatias Diabéticas/tratamento farmacológico , Matriz Extracelular/efeitos dos fármacos , Fator de Crescimento Transformador beta1/metabolismo , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo IV/metabolismo , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/metabolismo , Avaliação Pré-Clínica de Medicamentos , Células Epiteliais/efeitos dos fármacos , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Glucose/efeitos adversos , Humanos , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo , Transdução de Sinais/efeitos dos fármacos
15.
Cell Biol Int ; 35(12): 1239-41, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21675961

RESUMO

The present study aims to observe the effects of NCTD (norcantharidin) on proliferation and FN (fibronectin) expression in human renal proximal tubular epithelial cell line (HK-2) induced by albumin in vitro. HK-2 cells were divided into control group, albumin group and different concentration of NCTD intervention groups. Proliferation of HK-2 cells was determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide], FN protein in culture media of HK-2 cells was examined by ELISA, and FN mRNA was analysed by RT-PCR (reverse transcription-PCR). We chose less than 5.0 mg/l of NCTD as the experimental concentration for the cytotoxicity test. MTT score was higher in the albumin group than in the control group (P<0.05). As compared with that of the albumin group, MTT score and FN protein concentration decreased, FN mRNA significantly down-regulated in NCTD intervention groups respectively (P<0.05). Our study showed that NCTD could inhibit the albumin-induced cell proliferation and FN expression in HK-2 cells, which might further prove the anti-fibrotic role of NCTD in proteinuria-associated tubulointerstitial damage.


Assuntos
Albuminas/metabolismo , Antineoplásicos/farmacologia , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Proliferação de Células , Fibronectinas/genética , Linhagem Celular , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Fibronectinas/metabolismo , Humanos , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
16.
Ren Fail ; 33(2): 159-63, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21332337

RESUMO

The hallmark of IgA nephropathy (IgAN) is the mesangial deposits of polymeric IgA. However, the source of IgA1 and the mechanism of deposition of IgA1 in the mesangium remain unknown. To better understand its pathogenesis, we investigated the expression of CD19(+)CD5(+)B cells and IgA1-positive cells in the tonsils of IgAN patients. Immunofluorescence was used to visualize the locations of CD19(+)CD5(+)B cells and IgA1-positive cells in the tonsils. In this study, it was demonstrated that CD19(+)CD5(+)B cells are usually found in germinal centers and in the capsule covering the upper parts of the nodules of lymphoid tissue (cap of the nodule). The expression of IgA1-positive cells in tonsil tissue can be seen in the cap of the nodule and subepithelial tissue. There is a significant relationship between IgA1 and CD19(+)CD5(+)B cells. The level of CD19(+)CD5(+)B cells is positively correlated to the severity of renal pathological changes. These findings suggest that CD19(+)CD5(+)B cells in the tonsils could have an impact on the pathogenesis of IgAN.


Assuntos
Antígenos CD19/metabolismo , Linfócitos B/imunologia , Antígenos CD5/metabolismo , Glomerulonefrite por IGA/imunologia , Imunoglobulina A/metabolismo , Tonsila Palatina/imunologia , Adolescente , Adulto , Linfócitos B/metabolismo , Estudos de Casos e Controles , Criança , Feminino , Glomerulonefrite por IGA/patologia , Humanos , Rim/patologia , Masculino , Pessoa de Meia-Idade , Tonsila Palatina/metabolismo , Adulto Jovem
17.
Ren Fail ; 33(2): 233-41, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21332346

RESUMO

OBJECTIVE: To investigate the effects of norcantharidin (NCTD) on tubulointerstitial fibrosis of diabetic nephropathy (DN) in streptozotocin-induced rat model. METHODS: Sprague-Dawley rats were randomly divided into control group, model group, low-dose NCTD (0.05 mg/kg/day) group, and high-dose NCTD (0.1 mg/kg/day) group. The model group was induced by injection intraperitoneally with 30 mg/kg streptozotocin in 0.1 mol/L sodium citrate solution (pH 4.5), after high-calorie foods were given for 2 months. NCTD was administered daily after the DN rat model was built. Rats were sacrificed at the end of the third and the eighth week; renal fibrosis and the expression of FN, collagen IV, TGF-ß1, and calcineurin (CaN) were detected by Masson and immunohistochemistry staining, respectively. RESULTS: Tubulointerstitial fibrosis was observed in DN rats, this kind of pathological changes was ameliorated in NCTD treatment group (p < 0.05). The expressions of FN, collagen IV, and TGF-ß1 protein increased in the tubulointerstitial field of DN rats compared with the rats in control group. NCTD treatment could dose-dependently decrease their expression and reverse the fibrotic degree (p < 0.05). Meanwhile, the expression of CaN was detected in tubular fields of normal kidney and increased in the tubulointerstitial field in DN rats. However, NCTD downregulated its expression in a dose-dependent manner (p < 0.05). CONCLUSIONS: NCTD could downregulate FN, collagen IV, and TGF-ß1 expression in tubulointerstitial fields and attenuate tubulointerstitial fibrosis in the early stage of DN rats. NCTD also alleviated the expression of CaN in tubules in DN. The relationship between the role of NCTD's anti-tubulointerstitial fibrosis and its inhibition to CaN expression remains to be further elucidated.


Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/uso terapêutico , Nefropatias Diabéticas/tratamento farmacológico , Nefrite Intersticial/tratamento farmacológico , Animais , Calcineurina/metabolismo , Colágeno Tipo IV/metabolismo , Creatinina/sangue , Diabetes Mellitus Experimental/complicações , Nefropatias Diabéticas/induzido quimicamente , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Avaliação Pré-Clínica de Medicamentos , Fibronectinas/metabolismo , Fibrose , Rim/metabolismo , Rim/patologia , Masculino , Nefrite Intersticial/induzido quimicamente , Nefrite Intersticial/metabolismo , Nefrite Intersticial/patologia , Proteinúria/tratamento farmacológico , Proteinúria/etiologia , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1/metabolismo
18.
Med Hypotheses ; 75(6): 645-7, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20801582

RESUMO

Peritoneal dialysis has undergone considerable development from a technological point of view, and osmotic agent has played the essential role in peritoneal dialysis fluid. Because the most commonly used osmotic agent is glucose and icodextrin, there are some disadvantages related to the use of glucose-based solutions and icodextrin. So it is urgent to develop a new peritoneal dialysis osmotic agent. According to these characteristics of glucose and icodextrin, it is promising to explore a better osmotic agent of peritoneal dialysis solution which is able to allow maintenance of the maximum ultrafiltration gradient, and prevent toxicity or accumulation of unwanted substances in the blood, being non-toxic or less-toxic, furthermore the metabolite should not cause significant metabolic disturbance. Maltose may be one of promising osmotic agent and may put an important influence on development of peritoneal dialysis.


Assuntos
Soluções para Diálise/uso terapêutico , Maltose/uso terapêutico , Diálise Peritoneal/métodos , Soluções para Diálise/farmacologia , Humanos , Maltose/farmacologia , Osmose/efeitos dos fármacos
19.
Am J Physiol Renal Physiol ; 299(5): F1014-25, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20739391

RESUMO

p66Shc, a promoter of apoptosis, modulates oxidative stress response and cellular survival, but its role in the progression of diabetic nephropathy is relatively unknown. In this study, mechanisms by which p66Shc modulates high-glucose (HG)- or angiotensin (ANG) II-induced mitochondrial dysfunction were investigated in renal proximal tubular cells (HK-2 cells). Expression of p66Shc and its phosphorylated form (p-p66Shc, serine residue 36) and apoptosis were notably increased in renal tubules of diabetic mice, suggesting an increased reactive oxygen species production. In vitro, HG and ANG II led to an increased expression of total and p-p66Shc in HK-2 cells. These changes were accompanied with increased production of mitochondrial H(2)O(2), reduced mitochondrial membrane potential, increased translocation of mitochondrial cytochrome c from mitochondria into cytosol, upregulation of the expression of caspase-9, and ultimately reduced cell survival. Overexpression of a dominant-negative Ser36 mutant p66Shc (p66ShcS36A) or treatment of p66Shc- or PKC-ß-short interfering RNAs partially reversed these changes. Treatment of HK-2 cells with HG and ANG II also increased the protein-protein association between p-p66Shc and Pin1, an isomerase, in the cytosol, and with cytochrome c in the mitochondria. These interactions were partially disrupted with the treatment of PKC-ß inhibitor or Pin1-short interfering RNA. These data suggest that p66Shc mediates HG- and ANG II-induced mitochondrial dysfunctions via PKC-ß and Pin1-dependent pathways in renal tubular cells.


Assuntos
Angiotensina II/toxicidade , Apoptose/fisiologia , Glucose/toxicidade , Túbulos Renais/patologia , Mitocôndrias/fisiologia , Estresse Oxidativo/fisiologia , Proteínas Adaptadoras da Sinalização Shc/fisiologia , Animais , DNA Mitocondrial/biossíntese , Diabetes Mellitus Experimental/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Marcação In Situ das Extremidades Cortadas , L-Lactato Desidrogenase/metabolismo , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Microscopia Confocal , Peptidilprolil Isomerase de Interação com NIMA , Peptidilprolil Isomerase/metabolismo , Proteína Quinase C/metabolismo , Proteína Quinase C beta , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Proteínas Adaptadoras da Sinalização Shc/genética , Transdução de Sinais/fisiologia , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src
20.
Ther Apher Dial ; 14(1): 27-34, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20438517

RESUMO

Connective tissue growth factor (CTGF), a downstream mediator of transforming growth factor-beta1 (TGF-beta1) inducing fibrosis, has recently been implicated in peritoneal fibrosis. Extracellular matrix (ECM) accumulation and angiogenesis are characteristic changes in peritoneal fibrosis. In this study we investigated the effect of CTGF knockdown via interference RNA (RNAi) on ECM production and vascular endothelial growth factor (VEGF) expression induced by TGF-beta1 in human peritoneal mesothelial cells (HPMCs). Four CTGF short hairpin RNA (shRNA) expression constructs were generated using the pRetroSuper vector, and infectious retroviral particles were prepared to infect HPMCs. Expression levels of CTGF, fibronectin(FN), collagen 1 (col 1), laminin, and VEGF mRNA and protein were measured by semi-quantitative reverse transcription polymerase chain reaction and western blot assay. CTGF expression was increased after stimulation with TGF-beta1, but inhibited using each of the four independent CTGF shRNA constructs (P < 0.01). Moreover, expression of ECM proteins (FN, col 1, and laminin) and VEGF were upregulated after incubation with TGF-beta1, but elevated levels of ECM and VEGF induced by TGF-beta1 were significantly inhibited by RNAi (P < 0.01), but not by the empty retroviral vector (P > 0.05). From these results, we concluded that retrovirus-mediated CTGF shRNA can effectively inhibit ECM production and VEGF expression induced by TGF-beta1 in HPMCs. This study suggests that downregulation of CTGF may represent a potential therapeutic approach for peritoneal fibrosis through decreasing ECM accumulation and angiogenesis.


Assuntos
Fator de Crescimento do Tecido Conjuntivo/genética , Proteínas da Matriz Extracelular/genética , Técnicas de Silenciamento de Genes/métodos , Peritônio/metabolismo , Fator de Crescimento Transformador beta1/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Western Blotting/métodos , Células Cultivadas , Regulação para Baixo/genética , Epitélio , Proteínas da Matriz Extracelular/metabolismo , Expressão Gênica/genética , Humanos , Peritônio/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fator A de Crescimento do Endotélio Vascular/genética
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