引用

醛酰胺是由多种不同的植物产生的脂肪酸酰胺。alkamide具有免疫调节和抗痛觉作用，其作用机制的研究大多集中在alkamide对介导疼痛和炎症信号的受体和离子通道的作用上。alkamide是否能进入细胞并潜在地发挥细胞内作用是未知的，这将是本论文的主题。为了解决这个问题，我们通过用荧光素(FITC)分子取代天然产物(2E,4E)- n -异丁基十二烷基-2,4-二胺的异丁基头基，生产了荧光标记的阿尔卡酰胺。我们发现这种分子(FITC-Alk)保留了抑制TNF-产生的能力?来自lps刺激的RAW 264.7巨噬样细胞。然后使用共聚焦显微镜来描述FITC-Alk在RAW 264.7细胞中的进入和细胞内定位。我们的实验表明，FITC-Alk在几分钟内进入细胞，并在细胞质中形成大而明亮的点状和弥漫性针状染色。三维z层深度编码显示，在整个细胞质中以均匀的、非极化的方式形成点。在Vero绿猴肾细胞和RBL-2H3嗜碱性白血病细胞中也发现了相同的模式，这表明摄取的共同途径与细胞类型或起源组织无关。 Staining intensity was found to be both time and concentration dependent and continued linearly for at least 4 hours. Co-staining with Texas Red-dextran indicated that the large puncta produced by FITCAlk uptake were intracellular endocytic compartments. Additionally, our data suggest a role for actin filaments in FITC-Alk uptake, and that the FITC-Alk molecule likely accumulates in Rab7 positive, late-stage endosomes. Intracellular proteins should therefore be considered as targets for alkamide activity and alkamides may be useful for endocytic drug targeting.