RESUMO
Nuclear factor-erythroid 2-related factor 2 (Nrf2) is a transcription factor that regulates antioxidant and anti-inflammatory genes, and it plays a crucial role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Moreover, 15-deoxy-delta12,14-prostaglandin J2 (15d-PGJ2) plays a protective role against oxidative stress and inflammation both in vivo and in vitro. In a previous study, we found that 15d-PGJ2 increased the expression of Nrf2 in a COPD rat model. This study aims to elucidate the role of 15d-PGJ2 in COPD pathogenesis and the relationship between Nrf2 and human bronchial epithelial (HBE) cells. Normal HBE (HBE) cells were cultured. Following cigarette smoke extract (CSE) stimulation, pre-incubation with or without small interfering RNA (siRNA) Nrf2, and stimulation with or without 15d-PGJ2, the expression levels of Nrf2, NF-κBp65, and IL-8 were detected by reverse transcription-polymerase chain reaction and western blot, respectively. The expression of NF-κBp65 and IL-8 in CSE-stimulated normal HBE cells was inhibited by 15d-PGJ2 at both the mRNA level and the protein level. Moreover, the expression of Nrf2 in normal HBE cells was improved by 15d-PGJ2 at both the mRNA level and the protein level. However, the inhibitory or improving effects of 15d-PGJ2 were disengaged by siRNA Nrf2 at both the mRNA level and the protein level. 15d-PGJ2 possesses anti-inflammatory properties in the pathogenesis of COPD, and HBE cells stimulated by CSE via Nrf2 activation.
Assuntos
Anti-Inflamatórios/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Prostaglandina D2/análogos & derivados , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/metabolismo , Animais , Brônquios/citologia , Brônquios/efeitos dos fármacos , Brônquios/metabolismo , Células Cultivadas , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Modelos Animais , Fator 2 Relacionado a NF-E2/antagonistas & inibidores , Fator 2 Relacionado a NF-E2/biossíntese , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Prostaglandina D2/farmacologia , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/antagonistas & inibidores , Fator de Transcrição RelA/biossíntese , Fator de Transcrição RelA/metabolismoRESUMO
UNLABELLED: To explore the expression of interleukin-5(IL-5) and interleukin-10(IL-10) and their relationship in asthma, we measured the cells in bronchial alveolar lavage fluid(BALF) of guinea pigs, and examined the expression of IL-5 mRNA and IL-10 mRNA of bronchial tissue by reverse transcription polymerase chain reaction(RT-PCR) in control group(Group C, n = 10), asthmatic group(Group A, n = 11) and dexamethasone treatment group(Group B, n = 10). The results showed that the eosinophil(EOS) percentage and the expression of IL-5 mRNA were significantly higher in Group A[(48.64 +/- 17.8)%, (106.91 +/- 20.09)% respectively] than that in Group C[(15.10 +/- 11.48)%, (62.60 +/- 13.84)%] and Group B[(28.4 +/- 10.32)%, (75.30 +/- 10.70)%]. In contrast, the IL-10 mRNA expression in Group A[(47.5 +/- 5.3)%] was obviously lower compared with that in Group C[(101.2 +/- 4.2)%] and Group B[(80.1 +/- 8.3)%]. The correlation between the expression level of IL-5 mRNA and percentage of EOS was positive(r = 0.924, P < 0.01), whereas the IL-5 mRNA was negatively correlated with IL-10 mRNA expression(r = -0.731, P < 0.01). CONCLUSION: IL-5 and EOS might play a role in the pathogenesis of asthma, the inhibition of IL-10 mRNA expression may cause the increase of IL-5 expression in asthma. The beneficial effects of corticosteroid treatment in asthma may partly result from increasing IL-10 mRNA expression and inhibiting IL-5 mRNA expression.
Assuntos
Asma/metabolismo , Brônquios/metabolismo , Interleucina-10/biossíntese , Interleucina-5/biossíntese , Animais , Asma/patologia , Líquido da Lavagem Broncoalveolar/citologia , Eosinófilos/patologia , Feminino , Cobaias , Interleucina-10/genética , Interleucina-5/genética , Masculino , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Polymorphism of serum esterase was examined in 3 Super-Y 2000 broiler breeding populations differing in developmental stages (i.e. day 120, 180 and 300) using vertical polyacrylamide gel electrophoresis. The results indicated that both Es-1 and Es-2 loci exhibited polymorphism. Two to three polymorphic enzyme bands were detected at zone Es-1, and one band at zone Es-2 after electrophoresis. It was first discovered in this work that obvious developmental difference in products of Es-1 alleles existed with female birds, polymorphic bands of which disappeared after onset of lay. An individual tracing experiment conducted in a Super-Y 380 commercial layer population and an extensive experiment carried out in 2 ISA B380 parent CD stocks supported the previous finding. It could thus be inferred that phenotype "O" (no band at Es-1) discovered in hens was not genetically controlled by the recessive allele Es-1(0), which itself did not exist, but might be an evidence of gene regulation at laying period. For female birds, expression of Es-1 alleles was active before sexual maturity, and depressed after onset of lay, to meet their physiological need of high level of blood esterase for laying. It could also be inferred that this developmental difference in serum esterase polymorphism might be a common phenomenon in all avian species. If this hypothesis were true, the synthesis of esterase inside avian bodies could be a desirable model for investigating expression, and regulation of expression, of genes in avian populations.
