RESUMEN
Fibrotic diseases account for more than 8 million deaths worldwide annually. Reactive oxygen species (ROS) has been shown to activate pyroptosis and promote the production of interleukin (IL)-1ß and IL-18, leading to fibrosis development. However, the role of dual oxidase 1 (DUOX1)-induced ROS production and pyroptosis in cardiac fibrosis remains largely unknown. Activin A was used to induce ROS and pyroptosis in cardiomyocytes. ROS level, pyroptosis, and cytokine production were detected using Active Oxygen Detection Kit, flow cytometry, and enzyme-linked immunosorbent assay, respectively. Western blotting analysis was used to measure expression changes of proteins. DUOX1 was silenced or overexpressed to investigate its role in fibrosis. We found that activin A induced ROS production and pyroptosis in cardiomyocytes, which was blocked by the ROS scavenger, N-acetyl-L-cysteine (NAC). Knockdown of DUOX1 reversed activin A-induced ROS production, pyroptosis, cytokine release, and the upregulation of proinflammatory proteins. Overexpression of DUOX1 resulted in opposite effects of knockdown DUOX1. Administration of an ROS scavenger blocked the effect of DUOX1 overexpression. Supplementation of IL-1ß and IL-18 caused significant fibrosis in human cardiac fibroblasts (hCFs). The knockdown of DUOX1 protected cardiomyocytes against activin A-induced fibrosis via the inhibition of ROS, cytokine release, and pyroptosis.
Asunto(s)
Activinas/farmacología , Oxidasas Duales/genética , Miocitos Cardíacos/efectos de los fármacos , Piroptosis/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Acetilcisteína/farmacología , Activinas/antagonistas & inhibidores , Caspasa 1/genética , Caspasa 1/metabolismo , Coenzima A Ligasas/genética , Coenzima A Ligasas/metabolismo , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo III/genética , Colágeno Tipo III/metabolismo , Oxidasas Duales/antagonistas & inhibidores , Oxidasas Duales/metabolismo , Depuradores de Radicales Libres/farmacología , Regulación de la Expresión Génica , Humanos , Interleucina-18/genética , Interleucina-18/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Estrés Oxidativo/efectos de los fármacos , Cultivo Primario de Células , Piroptosis/genética , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Especies Reactivas de Oxígeno/agonistas , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Transducción de Señal , Proteína Smad2/genética , Proteína Smad2/metabolismo , Proteína smad3/genética , Proteína smad3/metabolismoRESUMEN
OBJECTIVE: To Analyze the chemical composition and their relative contents of essential oil from the leaves and fruits of Eucalyptus tereticornis in Guangxi province. METHODS: The column temperature was controlled by a program with a capillary column HP-5 MS, and the MS analysis was performed with EI and quadrupole mass analyzer. The chemical compositions were identified by NIST98 searching and mass spectra comparing, and their relative contents were determined by using normalization method of chromatographic peak areas. RESULTS: 39 compounds constituting 96.69% of the oil of leaves were identified. The major components were eucalyptol (27.93%), 1R-alpha-pinene (22.60%), isopinocarveol (8.71%); 36 compounds constituting 93.50% of the oil of fruits were identified. The major components are 1R-alpha-pinene (32.88%), eucalyptol (13.64%), D-limonene (8.31%). CONCLUSION: The GC-MS is a simple, rapid and sensitive method.