Intracellular distribution of NFkappaB decoy and its inhibitory effect on TNFalpha production by LPS stimulated RAW 264.7 cells.
Intracellular distribution of NFkappaB decoy and its inhibitory effect on TNFalpha production by LPS stimulated RAW 264.7 cells.
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发表时间：
2005
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通讯作者：
Y. Higuchi;S. Kawakami;M. Nishikawa;F. Yamashita;M. Hashida
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作者：
Y. Higuchi;S. Kawakami;M. Nishikawa;F. Yamashita;M. Hashida
核因子κ B （NFkappaB）是参与炎症性疾病发病机制的多种细胞因子表达的转录因子。未受刺激的NFkappaB被隔离在细胞质中，与抑制蛋白结合，称为IkappaBs。NFkappaB的许多激活因子导致IkappaB蛋白降解，游离的NFkappaB可以进入细胞核诱导基因表达。在本研究中，我们分析了NFkappaB诱饵在RAW 264.7细胞中的细胞内分布与药理作用的关系。大多数荧光标记的NFkappaB诱饵在没有LPS刺激或阳离子转染的细胞质中都可以观察到。此外，在LPS刺激下，细胞质中也观察到大部分NFkappaB诱饵。然而，NFkappaB诱饵有效地抑制了RAW 264.7细胞中TNFalpha的产生。阳离子脂质体转染NFkappaB诱饵对TNFalpha产生的抑制作用比裸NFkappaB诱饵强得多，因为阳离子脂质体转染NFkappaB诱饵的细胞关联量是裸NFkappaB诱饵的7倍。这些信息对NFkappaB诱饵载波系统的设计具有重要的参考价值。
Nuclear factor kappa B (NFkappaB) is a transcriptional factor for the expression of many cytokines that are involved in the pathogenesis of inflammatory diseases. Unstimulated NFkappaB sequestered in the cytoplasm bound to inhibitory proteins is called IkappaBs. Many activators of NFkappaB cause degradation of IkappaB proteins and free NFkappaB can enter the nucleus and induce gene expression. In this study, we analyzed the relationship between the intracellular distribution and pharmacological effect of NFkappaB decoy in RAW 264.7 cells. Most of the fluorescent labeled NFkappaB decoy was observed in the cytoplasm both with or without cationic transfection without LPS stimulation. Furthermore, under LPS stimulation, most of NFkappaB decoy was also observed in the cytoplasm. However, NFkappaB decoy effectively inhibited the production of TNFalpha in RAW 264.7 cells. The inhibitory effect of TNFalpha production by NFkappaB decoy transfected by cationic liposomes was much stronger than that by naked NFkappaB decoy, because the amount of cellular association of NFkappaB transfected by cationic liposome decoy was 7 times higher than that of naked NFkappaB decoy. This information is of great value for the design of NFkappaB decoy carrier systems.