Absorption Mechanism Of Cod Skin Collagen Peptide In Caco-2 Cell Monolayer Model
Keywords
Cod Skin, Collagen Peptides, Gastrointestinal Stability, Caco-2 Cells, Transport Mechanism
Abstract
Purpose: Cod skin collagen peptide (CSCP) has a good protective effect on liver damage, but its absorption mechanism is still unclear. This experiment intends to use the human colon adenocarcinoma Caco-2 cell monolayer model to treat CSCP. The absorption mechanism in the intestine was studied to provide basis for the absorption mechanism of CSCP in the intestine of animals. Methods: Before conducting the transport experiment, the stability of CSCP in artificial stomach, intestinal juice, different pH value conditions and in Caco-2 cell monolayer was first evaluated; the CCK-8 (cell counting kit-8) method was used to determine CSCP The highest mass concentration in the transport experiment, then establish a Caco-2 cell monolayer model and measure the transmembrane resistance value and alkaline phosphatase activity to examine the compactness, integrity and polarization of the cell monolayer; examine the transport time, Effects of CSCP mass concentration, verapamil, MK-571, phenyl arsenic oxide and sodium deoxycholate on transport. High performance liquid chromatography was used to detect CSCP mass concentration and calculate cumulative transport volume and apparent permeability coefficient. Results: Within 3 hours, CSCP remained basically stable in the artificial stomach, intestinal juice and environment with a pH value close to the gastrointestinal tract, and no hydrolysis of the polypeptide was seen during the transport process. The transport of CSCP is time- and concentration-dependent and is not affected by verapamil and phenylarsenic oxide. Sodium deoxycholate and MK-571 have a very significant promoting effect on the transport of CSCP (P£¼0.05). Conclusion: The mechanism of CSCP transport across Caco-2 cell monolayer is the paracellular pathway, and its efflux is mediated by multidrug resistance proteins.
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Original research was done by Ding Guofang, Yu Fangmiao, Tang Yunping, Zhang Xiaojun, Yang Zuisu, Mei Guangming, Chen Si, Chen Rui, Huang Fangfang
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