Transport of Antihypertensive Peptide RVPSL, Ovotransferrin 328-332, in Human Intestinal Caco-2 Cell Monolayers
Transport of Antihypertensive Peptide RVPSL, Ovotransferrin 328-332, in Human Intestinal Caco-2 Cell Monolayers
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DOI：
10.1021/acs.jafc.5b01824
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发表时间：
2015-09-23
影响因子：
6.1
通讯作者：
Liu, Jingbo
中科院分区：
文献类型：
--
作者：
Ding, Long;Wang, Liying;Liu, Jingbo
本研究的目的是研究具有血管紧张素转换酶（ACE）抑制和抗高血压活性的蛋清衍生肽RVPSL（Arg-Val-Pro-Ser-Leu）在人肠Caco-2细胞单层中的跨上皮转运。结果表明，RVPSL可以被动转运通过Caco-2细胞单层。然而，在转运过程中，36.31% +/- 1.22%的初始RVPSL被降解，但当Caco-2细胞单层与diprotin A预孵育时，这种降解降低到23.49% +/- 0.68%（P < 0.001），表明RVPSL对各种刷状缘膜肽酶具有低抗性。当研究从顶侧至基底侧的转运时，表观渗透系数（P-app）为（6.97 +/- 1.11）x 10（-6）cm/s。RVSL的转运途径似乎是通过紧密连接的细胞旁途径，因为只有细胞松弛素D（一种紧密连接（TJ）的破坏剂）显著增加转运速率（P < 0.001）。此外，通过对RVPSL的突变，研究了RVPSL结构与跨Caco-2细胞单层转运的关系。发现N-末端Pro残基比Arg和瓦尔更有利于五肽跨Caco-2细胞单层的转运。此外，RVPSL可以更容易地作为较小的肽，特别是在二肽和三肽的形式进行转运。
The objective of this study was to investigate the transepithelial transport of RVPSL (Arg-Val-Pro-Ser-Leu), an egg-white-derived peptide with angiotensin I-converting enzyme (ACE) inhibitory and antihypertensive activity, in human intestinal Caco-2 cell monolayers. Results revealed that RVPSL could be passively transported across Caco-2 cell monolayers. However, during the process of transport, 36.31% +/- 1.22% of the initial RVPSL added to the apical side was degraded, but this degradation decreased to 23.49% +/- 0.68% when the Caco-2 cell monolayers were preincubated with diprotin A (P < 0.001), suggesting that RVPSL had a low resistance to various brush border membrane peptidases. When transport from the apical side to the basolateral side was investigated, the apparent permeability coefficient (P-app) was (6.97 +/- 1.11) x 10(-6) cm/s. The transport route of RVPSL appears to be the paracellular pathway via tight junctions, as only cytochalasin D, a disruptor of tight junctions (TJs), significantly increased the transport rate (P < 0.001). In addition, the relationship between the structure of RVPSL and transport across Caco-2 cell monolayers was studied by mutation of RVPSL. It was found that N-terminal Pro residues were more beneficial for transport of pentapeptides across Caco-2 cell monolayers than Arg and Val. Furthermore, RVPSL could be more easily transported as smaller peptides, especially in the form of dipeptides and tripeptides.