RESUMO
The molecular mechanism of DNA damage induced by hydroquinone (HQ) remains unclear. Poly(ADP-ribose) polymerase-1 (PARP-1) usually works as a DNA damage sensor, and hence, it is possible that PARP-1 is involved in the DNA damage response induced by HQ. In TK6 cells treated with HQ, PARP activity as well as the expression of apoptosis antagonizing transcription factor (AATF), PARP-1, and phosphorylated H2AX (γ-H2AX) were maximum at 0.5 h, 6 h, 3 h, and 3 h, respectively. To explore the detailed mechanisms underlying the prompt DNA repair reaction, the above indicators were investigated in PARP-1-silenced cells. PARP activity and expression of AATF and PARP-1 decreased to 36%, 32%, and 33%, respectively, in the cells; however, γ-H2AX expression increased to 265%. Co-immunoprecipitation (co-IP) assays were employed to determine whether PARP-1 and AATF formed protein complexes. The interaction between these proteins together with the results from IP assays and confocal microscopy indicated that poly(ADP-ribosyl)ation (PARylation) regulated AATF expression. In conclusion, PARP-1 was involved in the DNA damage repair induced by HQ via increasing the accumulation of AATF through PARylation.
Assuntos
Antioxidantes/toxicidade , Proteínas Reguladoras de Apoptose/metabolismo , Dano ao DNA/efeitos dos fármacos , Hidroquinonas/toxicidade , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Repressoras/metabolismo , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Inativação Gênica , Histonas/genética , Histonas/metabolismo , Humanos , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/genética , Transporte Proteico , Proteínas Repressoras/genéticaRESUMO
This study aimed to examine the number of circulating Toll-like receptor 4 (TLR4) + CD14+ monocytes in patients with different stages of chronic kidney disease (CKD), their responses to lipopolysaccharide (LPS), and to explore the potential association of the number of TLR4+CD14+ monocytes with clinical laboratory measures. The numbers of TLR4+CD14+, LPS-stimulated TNF-α+CD14+ and interleukin (IL)-6+CD14+ monocytes were determined by flow cytometry in 9 patients with stage 3 CKD, 11 with stage 4 CKD, 16 with stage 5 CKD, and 19 healthy controls (HCs). Their laboratory tests were performed by routine methods and the potential association among these measures was analyzed by Pearson's correlation analysis. The numbers of CD14+, CD14+TLR4+, LPSstimulated TNF-α+CD14+ and IL-6+CD14+ monocytes in patients with CKD were significantly less than those of HCs (all P<0.05), and were negatively associated with patient disease severity. The number of CD14+TLR4+ monocytes was positively correlated with estimated glomerular filtration rate (eGFR, P<0.001) and the levels of hematocrit (P<0.01), but negatively correlated with the levels of blood urine nitrogen, serum creatinine, and C-reactive protein (P<0.001 for all), in the CKD patients. Our data indicate that significant reduction in the number of TLR4+ monocytes and their impaired responses to LPS may be associated with the progression of CKD in Chinese patients.
Assuntos
Citocinas/metabolismo , Falência Renal Crônica/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/farmacologia , Monócitos/metabolismo , Receptor 4 Toll-Like/metabolismo , HumanosRESUMO
The protective effect of erythropoietin (EPO) on tissues following ischemia and reperfusion injuries remains poorly understood. We aimed to investigate the effect of EPO in preventing endotoxin-induced organ damage. Rat model of multiple organ failure (MOF) was established by tail vein injection of 10 mg/kg lipopolysaccharide (LPS). Recombinant human EPO treatment (5000 U/kg) was administered by tail vein injection at 30 min after LPS challenge. Twenty-four h after EPO treatment, changes in serum enzyme levels, including aspartate aminotransferase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN) and creatinine (Cr), were evaluated by biochemical analysis. Serum levels of tumor necrosis factor-α (TNF-α) were determined by using immunoradiometric assay. Histological examination of tissue sections was carried out by hematoxylin and eosin staining, while ultrastructure evaluation of organ tissues was assessed by transmission electron microscopy. Protein expression levels were detected by using Western blotting. EPO treatment showed a modest effect in preventing LPS-induced elevation of AST, ALT, BUN, Cr, and TNF-α levels, and in protecting against LPS-induced tissue degeneration and injured ultrastructure in the lung, liver, and kidney. Moreover, LPS promoted phosphorylation of alanine aminotransferase (AKT) and increased nuclear factor-κB (NF-κB) activation in the lung, liver, and kidney (P<0.05 vs. control). However, EPO treatment significantly decreased the LPS-induced pAKT up-regulation in these tissues (P<0.05 vs. LPS treatment alone). The present study demonstrates that EPO may play a protective role against LPS-induced MOF by reducing the inflammatory response and tissue degeneration, possibly via the phosphatidylinositol 3-kinase/AKT and NF-κB signaling pathways.
Assuntos
Eritropoetina/farmacologia , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Insuficiência de Múltiplos Órgãos/prevenção & controle , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Nitrogênio da Ureia Sanguínea , Western Blotting , Creatinina/sangue , Endotoxinas , Eritropoetina/administração & dosagem , Eritropoetina/genética , Injeções Intravenosas , Rim/metabolismo , Rim/ultraestrutura , Lipopolissacarídeos , Fígado/metabolismo , Fígado/ultraestrutura , Pulmão/metabolismo , Pulmão/ultraestrutura , Masculino , Microscopia Eletrônica de Transmissão , Insuficiência de Múltiplos Órgãos/sangue , Insuficiência de Múltiplos Órgãos/induzido quimicamente , NF-kappa B/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Wistar , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Fator de Necrose Tumoral alfa/sangueRESUMO
To explore the characteristics and relevant risk factors of periodontal disease (PD) among hemodialysis patients.Uremic patients on maintenance hemodialysis from November 2015 to March 2016 were retrospectively reviewed. Patients were divided into a PD group and a non-PD group. Demographic and laboratory data were collected and analyzed.In all, 136 uremic patients (79 males and 57 females, aged 50.8â±â15.3 years) on maintenance hemodialysis were included in this study. The incidence of PD increased with age. Hemodialysis patients most likely developed PD if they were male, smokers, or diabetic (Pâ=â.009, <.001, and <.001, respectively). Patients brushing their teeth twice daily had significantly less chance of developing PD as compared with those only brushing once daily (Pâ<â.001). Hemodialysis patients in the PD group had significantly higher levels of total cholesterol, high-sensitivity C-reactive protein, fasting blood glucose, and peripheral white blood cell counts, compared with the non-PD group (all Pâ<â.001). Logistic regression analysis revealed that diabetes, total cholesterol, high-sensitivity C-reactive protein, and peripheral white blood cell count were independent risk factors for developing PD, whereas teeth brushing twice daily and serum calcium were favorable factors for maintenance hemodialysis patients against PD.Identification of risk factors provides a theoretical basis for prevention and improvement of PD among maintenance hemodialysis patients.
Assuntos
Doenças Periodontais/epidemiologia , Diálise Renal/estatística & dados numéricos , Insuficiência Renal Crônica/epidemiologia , Adulto , Idoso , Glicemia , Proteína C-Reativa/análise , Colesterol/sangue , Diabetes Mellitus/epidemiologia , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Higiene Bucal , Insuficiência Renal Crônica/terapia , Estudos Retrospectivos , Fatores de Risco , Fatores Sexuais , Fumar/epidemiologiaRESUMO
We aimed to investigate the differences in renal histopathological changes and laboratory parameters between adult and pediatric patients with Henoch-Schönlein purpura nephritis (HSPN), and to analyze the correlation between laboratory parameters and renal histopathological grading. A total of 139 patients diagnosed with HSPN between September 2010 and December 2014 at the First Hospital of Jilin University, China, were retrospectively reviewed. The clinical and pathological characteristics were examined and compared between the adult and the pediatric patients. A majority of adult (75.0%) and pediatric (66.2%) patients were categorized as pathological grade III HSPN. Adults having crescent lesions, interstitial fibrosis and renal artery involvement significantly outnumbered child counterparts (all P<0.05). Pathological grading showed a positive correlation with 24-h urine protein (r=0.307, P=0.009), microalbuminuria (r=0.266, P=0.000) and serum globulin (r=0.307, P=0.014), and a negative correlation with serum albumin (r=0.249, P=0.037) in pediatric patients with HSPN. Among adult patients with HSPN, histopathological grading showed a positive correlation with 24-h urine protein (r=0.294, P=0.015), microalbuminuria (r=0.352, P=0.006), α1-microglobulin (r=0.311, P=0.019) and immunoglobulin G (r=0.301, P=0.023) in urine, and serum creatinine (r=0.292, P=0.018). Further, a negative correlation between serum albumin and pathological grading was also observed (r=0.291, P=0.018). In conclusion, the severity of renal pathological lesions in HSPN patients is well reflected by the levels of proteinuria. Adult patients have more severe renal histopathological changes than pediatric patients.
Assuntos
Vasculite por IgA/sangue , Vasculite por IgA/urina , Nefrite/sangue , Nefrite/urina , Adolescente , Adulto , Criança , Pré-Escolar , China , Creatinina/sangue , Feminino , Humanos , Vasculite por IgA/fisiopatologia , Imunoglobulina G/urina , Masculino , Nefrite/fisiopatologia , Proteinúria/metabolismo , Proteinúria/fisiopatologia , Albumina Sérica/metabolismoRESUMO
The effectiveness of ulinastatin and methylprednisolone in treating pathological changes in mice with radiation-induced lung injury (RILI) was evaluated. Forty C57BL/6 female mice received whole-chest radiation (1.5 Gy/min for 12 min) and were randomly allocated into Group R (single radiation, n = 10), Group U (ulinastatin treatment, n = 10), Group M (methylprednisolone treatment, n = 10), or Group UM (ulinastatin and methylprednisolone treatment, n = 10). Another 10 untreated mice served as controls (Group C). Pathological changes in lung tissue, pulmonary interstitial area density (PIAD) and expression levels of transforming growth factor ß1 (TGF-ß1) and tumor necrosis factor α (TNF-α) in lung tissue, serum and bronchoalveolar lavage fluid were determined. Alleviation of pathological changes in lung tissue was observed in Groups U, M and UM. Treatment with ulinastatin, methylprednisolone or both effectively delayed the development of fibrosis at 12 weeks after radiation. Ulinastatin, methylprednisolone or both could alleviate the radiation-induced increase in the PIAD (P < 0.05 or P < 0.01). Treatment with ulinastatin, methylprednisolone or both significantly reduced the expression of TNF-α, but not TGF-ß1, at 9 weeks after radiation compared with Group R (P < 0.01). Ulinastatin and /: or methylprednisolone effectively decreased the level of TNF-α in lung tissue after RILI and inhibited both the inflammatory response and the development of fibrosis.
Assuntos
Glicoproteínas/uso terapêutico , Lesão Pulmonar/tratamento farmacológico , Lesão Pulmonar/etiologia , Metilprednisolona/uso terapêutico , Lesões por Radiação/tratamento farmacológico , Protetores contra Radiação/uso terapêutico , Animais , Líquido da Lavagem Broncoalveolar , Feminino , Glicoproteínas/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Lesão Pulmonar/sangue , Metilprednisolona/farmacologia , Camundongos Endogâmicos C57BL , Lesões por Radiação/sangue , Fator de Crescimento Transformador beta1/sangue , Fator de Necrose Tumoral alfa/sangueRESUMO
Omentin is a novel adipokine, which is expressed in and released from omental adipose tissue. In the present study, the effect of omentin on neural stem cells (NSCs) was investigated. NSCs are a subtype of stem cell in the nervous system, which are able to selfrenew and generate neurons and glia for repairing neural lesions. Mouse NSCs were isolated and cultured in vitro. Treatment with recombinant omentin for 3 and 5 days significantly increased the size of NSC neurospheres (P<0.01) and enhanced NSC cell viability in normal conditions. In addition, omentin protected against the decrease in cell viability induced by the proinflammatory cytokine tumor necrosis factorα. In the NSCs, incubation of omentin for 2, 4, 6, 8 and 16 h enhanced the phosphorylation of Akt at the Thr308 site and of AS160 at the Ser318 site, peaking 6 h after treatment. Additionally, treatment with LY294002 (10 µM), a specific inhibitor of phosphatidylinositol 3kinase/Akt signaling, eliminated the omentininduced increase in neurosphere size and cell viability. Overall, the present study provided the first evidence, to the best of our knowledge, that omentin promotes the growth and survival of NSCs in vitro through activation of the Akt signaling pathway. These results may contribute to the understanding of the role of omentin in the nervous system.
Assuntos
Citocinas/farmacologia , Lectinas/farmacologia , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Proteínas Ligadas por GPI/farmacologia , Humanos , Camundongos , Estresse FisiológicoRESUMO
Endothelial progenitor cells (EPCs), a group of bone marrow-derived pro-angiogenic cells, contribute to vascular repair after damage. EPC dysfunction exists in diabetes and results in poor wound healing in diabetic patients with trauma or surgery. The aim of the present study was to determine the effect of quercetin, a natural flavonoid on high glucoseinduced damage in EPCs. Treatment with high glucose (40 mM) decreased cell viability and migration, and increased oxidant stress, as was evidenced by the elevated levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase in bone marrow-derived EPCs. Moreover, high glucose reduced the levels of endothelial nitric oxide synthase (eNOS) phosphorylation, nitric oxide (NO) production and intracellular cyclic guanosine monophosphate (cGMP). Quercetin supplement protected against high glucoseinduced impairment in cell viability, migration, oxidant stress, eNOS phosphorylation, NO production and cGMP levels. Quercetin also increased Sirt1 expression in EPCs. Inhibition of Sirt1 by a chemical antagonist sirtinol abolished the protective effect of quercetin on eNOS phosphorylation, NO production and cGMP levels following high glucose stress. To the best of our knowledge, the results provide the first evidence that quercetin protects against high glucoseinduced damage by inducing Sirt1-dependent eNOS upregulation in EPCs, and suggest that quercetin is a promising therapeutic agent for diabetic patients undergoing surgery or other invasive procedures.
Assuntos
Células da Medula Óssea/patologia , Citoproteção/efeitos dos fármacos , Células Progenitoras Endoteliais/patologia , Glucose/toxicidade , Quercetina/farmacologia , Animais , Células Progenitoras Endoteliais/efeitos dos fármacos , Células Progenitoras Endoteliais/enzimologia , Humanos , Masculino , Camundongos Endogâmicos C57BL , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Sirtuína 1/metabolismoRESUMO
Heat-insoluble cryoglobulinemia is rare, and its pathogenesis and comorbidities remain poorly understood. Here, the authors report a case of hepatitis C virus (HCV)-related heat-insoluble cryoglobulinemia associated with thrombotic microangiopathy and cryoglobulin-occlusive membranoproliferative glomerulonephritis. The patient, a 57-year-old woman, presented with acute kidney injury, thrombocytopenia, anemia with schistocytes, high levels of serum HCV RNA of HCV genotype 2a, rheumatoid factor positivity and high levels of serum immunoglobulin (Ig) M and Igκ. The patient's serum was positive for cryoglobulin at 4°C, and the precipitate required heating to 47°C for dissolution. Cryoglobulin immunofixation was positive for monoclonal IgM and Igκ and polyclonal IgG. However, immunofixation of the cryoglobulin supernatant was negative. Histological examination of renal biopsy revealed a membranoproliferative type I glomerulonephritis. The patient was treated with plasmapheresis, corticosteroids and antiviral therapy of peginterferon plus ribavirin, but symptoms only partially resolved.
Assuntos
Crioglobulinemia/diagnóstico , Glomerulonefrite Membranoproliferativa/diagnóstico , Hepatite C/diagnóstico , Microangiopatias Trombóticas/diagnóstico , Anti-Inflamatórios/uso terapêutico , Antivirais/uso terapêutico , Análise Química do Sangue , China , Crioglobulinemia/complicações , Crioglobulinemia/tratamento farmacológico , Crioglobulinas/metabolismo , Feminino , Glomerulonefrite Membranoproliferativa/complicações , Glomerulonefrite Membranoproliferativa/tratamento farmacológico , Hepacivirus/isolamento & purificação , Hepatite C/complicações , Hepatite C/tratamento farmacológico , Hepatite C/virologia , Temperatura Alta , Humanos , Interferon alfa-2 , Interferon-alfa/uso terapêutico , Metilprednisolona/uso terapêutico , Pessoa de Meia-Idade , Plasmaferese , Polietilenoglicóis/uso terapêutico , Proteínas Recombinantes/uso terapêutico , Ribavirina/uso terapêutico , Microangiopatias Trombóticas/etiologiaRESUMO
Erythropoietin (EPO) has protective effects on myocardial infarction (MI). This study was to test whether EPO administrated after MI is as effective as EPO administrated before MI and to determine the role of AMP-activated protein kinase (AMPK) in the cardioprotective effects of EPO. Recombinant human EPO (5000 IU/kg) was intraperitoneally injected 12 hours before MI (pretreatment) or 30 minutes after MI (posttreatment). The levels of serum enzymes were assayed at 24 and 72 hours after MI. The infarct size was determined by nitro blue tetrazolium staining. The microarchitecture damages were evaluated by electron microscopy. EPO receptor messenger RNA and protein levels were determined by RT-PCR and immunohistochemistry, respectively. Activation of AMPK and nuclear factor kappa B was determined by immunoblotting. An AMPK-specific inhibitor, compound C, was used to inhibit AMPK activation in vivo. The authors found that both pretreatment and posttreatment of EPO successfully attenuated the serum enzyme levels, reduced the infarct size and ameliorated the microarchitecture damage. Moreover, both pretreatment and posttreatment of EPO decreased the EPO receptor messenger RNA and protein expressions, which were up-regulated in MI. More importantly, under MI conditions, EPO further increased the phosphorylation of AMPK and suppressed the activation of nuclear factor kappa B. Moreover, when AMPK was blocked by compound C, the cardioprotective effect of EPO was significantly attenuated (P < 0.01). Thus, this study demonstrates that both pretreatment and posttreatment of EPO are cardioprotective in an AMPK-dependent manners, providing the first evidence that the AMPK signaling pathway is involved in the cardioprotective effects of EPO.