RESUMO
OBJECTIVE: To investigate the expression of inhibitory kappa B alpha (IkappaBalpha) and it's regulatory effect on nuclear factor kappa B (NF-kappaB) during viral gene transactivation triggered by respiratory tract viruses in steroid responsive simple nephrotic syndrome (SRSNS). METHODS: Children with SRSNS (including at active and remissive stage) were examined for this research project, and healthy children were chosen as controls. TaqMan reverse transcription-PCR (RT-PCR), Western blot, electro-mobility shift assays were performed to study the protein expression of IkappaBalpha gene and the activity of NF-kappaB in peripheral blood mononuclear cells (PBMCs). Simultaneously, RT-PCR was used to detect the gene expression of respiratory tract viruses (including respiratory syncytial virus and influenza virus), and viral antibody was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Comparing with that from SRSNS at remissive stage and healthy children, the activity of NF-kappaB from SRSNS at active stage was much higher statistically (P < 0.05). The positive correlation between NF-kappaB activation and the gene expression of respiratory tract viruses was observed (OR = 27, Kappa coefficient = 0.59, P <0. 05). In addition, levels of IkappaBalpha protein from children with SRSNS at active stage were much lower than those from healthy children and those from children with SRSNS at remissive stage (P < 0.05). There was a linear negative correlation between the activity of NF-kappaB and the protein level of IkappaBalpha (r = -0.884, P < 0.05). On the other hand, levels of IkappaBalpha mRNA from SRSNS at active stage were much higher than those from healthy children, but much lower than those from children with SRSNS at remissive stage (P < 0.05). CONCLUSION: Abnormal expression of IkappaBalpha, which is attributed to regulate activity of NF-kappaB, occurs during viral gene transactivation triggered by respiratory tract viruses in SRSNS.
Assuntos
Genes Virais/genética , Proteínas I-kappa B/metabolismo , NF-kappa B/metabolismo , Síndrome Nefrótica/genética , Esteroides/farmacologia , Ativação Transcricional , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Humanos , Proteínas I-kappa B/genética , Lactente , Masculino , Inibidor de NF-kappaB alfa , Síndrome Nefrótica/tratamento farmacológico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sistema Respiratório/virologia , Esteroides/uso terapêuticoRESUMO
OBJECTIVE: To investigate the path of viral transactivation of transcription by nuclear factor-kappa B (NF-kappaB) in the pathogenesis of steroid-responsive simple nephrotic syndrome (SRSNS) triggered by respiratory tract viruses. METHODS: Children with SRSNS (including active stage, convalescent and remissive stage) were examined and were analytically compared to children with nephritic nephrosis, secondary glomerular diseases and bronchiolitis; healthy children were enrolled as controls. The methods of electrophoretic mobility shift assay (EMSA), of reverse transcription-PCR (RT-PCR), and of enzyme-linked immunosorbent assay (ELISA), were used to detect the transcriptive activity of NF-kappaB, the gene expression of respiratory tract viruses (respiratory syncytial virus-RSV, influenza virus-Flu) in peripheral blood mononuclear cells (PBMC), and the levels of IL-8 and viral antibodyies in serum, respectively. RESULTS: (1) Compared with the control and SRSNS in convalescent and remissive stage, the activity of NF-kappaB in PBMC of SRSNS in active stage was statistically elevated (P<0.05). (2)The level of IL-8 in serum of SRSNS in active stage was significantly increased, compared with healthy control (P<0.001). There was a positive linear correlation between the activity of NF-kappaB and the level of IL-8 in active stage of SRSNS, and a positive correlation trend was observed between the activity of NF-kappaB and the gene expression of respiratory tract viruses. CONCLUSION: Viral transactivation of transcription by NF-kappaB may be an important path for respiratory tract viruses triggering the pathogensis of SRSNS.