An experimental study of the inhibitory effects on the activation of endotoxin-induced Kupffer cells through short hairpin RNA targeting interleukin-1 receptor associated kinase-4 gene / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 819-822, 2005.
Artículo
en Chino
| WPRIM
| ID: wpr-276342
ABSTRACT
<p><b>OBJECTIVE</b>To explore the inhibitory effects on the activation of endotoxin-induced Kupffer cells (KCs) through short hairpin RNA (shRNA) targeting interleukin-1 receptor associated kinase-4 (IRAK-4) gene.</p><p><b>METHODS</b>Two effective transfection shRNA plasmid (pSIIRAK-4-A, pSIIRAK-4-B) and one invalidated plasmids (pSIIRAK-4-C) targeting IRAK-4 gene were constructed. The isolated mouse KCs were divided into three groups the normal control group, the RNAi control group (pSIIRAK-4-C) and the RNAi effective group (pSIIRAK-4-A, pSIIRAK-4-B). Then KCs were stimulated with 0.1 microg/ml lipopolysaccharide (LPS) after 24 h transfection with the constructed plasmid. The expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blot at 6 h after LPS stimulation, and the activities of NF-kappaB in KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h.</p><p><b>RESULTS</b>The level of IRAK-4, the activities of NF-kappaB and the TNF-alpha level in the RNAi effective group were evidently lower than those in normal and RNAi control groups (P < 0.01) at 1 h, 3 h, and 6 h. Especially, the pSIIRAK-4-A group in which the changes of the above indices were of no difference (P > 0.05), had better inhibited effects than that of the pSIIRAK-4-B group (P < 0.01).</p><p><b>CONCLUSION</b>The shRNA targeting IRAK-4 gene could effectively inhibit the activation of endotoxin-induced KCs.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Fisiología
/
Transducción de Señal
/
ARN Interferente Pequeño
/
Interferencia de ARN
/
Endotoxinas
/
Quinasas Asociadas a Receptores de Interleucina-1
/
Genética
/
Macrófagos del Hígado
/
Metabolismo
/
Ratones Endogámicos BALB C
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Hepatology
Año:
2005
Tipo del documento:
Artículo
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