Zebrafish: A Visual Model To Evaluate the Biofate of Transferrin Receptor-Targeted 7Peptide-Decorated Coumarin 6 Micelles
Zebrafish: A Visual Model To Evaluate the Biofate of Transferrin Receptor-Targeted 7Peptide-Decorated Coumarin 6 Micelles
复制标题
斑马鱼：评估转铁蛋白受体靶向 7 肽修饰香豆素 6 胶束生物命运的视觉模型
DOI：
10.1021/acsami.7b12809
复制
发表时间：
2017-11-08
影响因子：
9.5
通讯作者：
Zheng, Ying
中科院分区：
文献类型：
--
作者：
Li, Ye;Song, Xiaoning;Zheng, Ying
在本研究中，斑马鱼被用作一种体内模型，以评估转铁蛋白受体（TfR）靶向的香豆素6（C6）胶束在各种生物屏障中的生物命运。构建了三种负载荧光香豆素6（7pep - M - C6）且配体密度不同的由七肽（7pep）修饰的聚（乙二醇） - 嵌段 - 聚（ε - 己内酯）胶束，其粒径在30到40纳米之间。整体免疫染色显示，视网膜、大脑和肠道中TfR的表达水平随着发育阶段的增加而升高。与未修饰的胶束相比，7pep - M - C6在斑马鱼幼鱼中表现出更高的摄取效率。用7pep对TfR进行预抑制表明，在7pep - M - C6的摄取过程中涉及TfR介导的内吞途径。斑马鱼幼鱼眼睛和大脑的共聚焦图像分别显示，C6能有效地穿过视网膜色素上皮层到达神经节细胞层，以及在所有脑组织中显著积累，且当配体密度为10%时达到平台期。为了研究C6在肠道中的分布，通过灌胃法将胶束给予成年斑马鱼。值得注意的是，7pep - M - C6增强了C6穿过绒毛的运输，并增加了其在肠道基底外侧膜的聚集。在口服7pep - M - C6后，C6在眼睛和大脑中积累。福斯特共振能量转移分析表明，在成年斑马鱼口服给药后，完整的7pep修饰的胶束能够进入肠道、大脑和眼睛的上皮细胞。总之，斑马鱼可作为一种模型，用于对TfR靶向给药系统的生物命运进行体内可视化评估。
In the present study, the zebrafish was explored as an in vivo model to assess the biofate of transferrin receptor (TfR)-targeted coumarin 6 (C6) micelles across various biological barriers. Three 7peptide (7pep)-decorated poly-(ethylene glycol)-block-poly(epsilon-caprolactone) micelles loaded with fluorescence coumarin 6 (7pep-M-C6) with different ligand densities were constructed with particle sizes between 30 and 40 nm. Whole-mount immunostaining revealed that the expression level of TfR. in the retina, brain, and intestine increased along with development stage. Compared to unmodified micelles, 7pep-M-C6 demonstrated higher uptake efficiency in the larval zebrafish. Preinhibition of TfR with 7pep implicated the TfR-mediated endocytosis pathway in the uptake of 7pep-M-C6. Confocal images of the larval zebrafish eye and brain showed the efficient delivery of C6 across the retinal pigment epithelial to the ganglion cell layer and the significant accumulation of C6 in all brain tissues, respectively, which plateaued when the ligand density was 10%. To investigate the intestinal distribution of C6, micelles were administered to adult zebrafish via gavaging. Notably, 7pep-M-C6 enhanced the transport of C6 across the villi and increased its aggregation into the basolateral membrane of the intestine. After the oral administration of 7pep-M-C6, C6 accumulated in the eye and brain. Forster resonance energy transfer analysis suggested thatintact 7pep-modified micelles could enter the epithelial cells of the intestine, brain, and eye after oral administration in adult zebrafish. In conclusion, zebrafish could be used as a model for in vivo visual assessment of the biofate of TfR-targeted drug delivery systems.