花蝇吻鲍是一种高蛋白食品，在亚洲国家广泛食用。它已知具有抗氧化、抗癌和抗菌作用。由于花蝇吻鲍血淋巴的生物学重要性尚未得到广泛研究，本研究的目的是纯化酚氧化酶（PO）并研究其对人结肠上皮细胞的免疫效应。PO通过硫酸铵沉淀和一步色谱柱层析法纯化。蛋白质的分子量约为270 kDa。当PO与革兰氏阴性细菌源脂多糖（LPS）以不同比例混合（10:1-1:10，w/w）时，剩余LPS的量减少。浓度高达200 μg/mL的PO对HT-29细胞无细胞毒性。当PO的浓度增加时，LPS在HT-29细胞中诱导的炎症反应被调节。随着PO浓度的增加，生产前炎症细胞因子、与高免疫反应相关的细胞因子（如IL4、IL-5和INF-γ）和前列腺素2（PGE2）的水平被调节。本研究认为，PO和LPS同时处理，通过调节ERK1/2介导的NF-κB通路在HT-29细胞中表现出抗炎作用。本研究的结果表明, 花蝇吻鲍血淋巴液参与了调节人结肠上皮细胞中革兰氏阴性菌相关的炎症免疫反应。版权所有©2023年Elsevier Ltd.发表。

Haliotis discus hannai, a food with a high protein content, is widely consumed in Asian countries. It is known to have antioxidant, anticancer, and antibacterial effects. Since the biological significance of H. discus hannai hemolymph has not been widely studied, the objective of the present study was to purify phenoloxidase (PO) and investigate its immunological effects on human colonic epithelial cells. PO was purified through ammonium sulfate precipitation and one step column chromatography. The molecular weight of the protein was about 270 kDa. When PO was mixed with Gram-negative bacteria-derived lipopolysaccharide (LPS) at various ratios (10:1-1:10, w/w), the amount of residual LPS was reduced. PO at concentrations up to 200 μg/mL was not cytotoxic to HT-29 cells. The inflammatory response induced by LPS in HT-29 cells was regulated when the concentration of PO was increased. With increasing concentration of PO, production levels of pro-inflammatory cytokines, cytokines associated with hyperimmune responses such as IL4, IL-5, and INF-γ, and prostaglandin 2 (PGE2) were regulated. It was thought that simultaneous treatment with PO and LPS anti-inflammatory effects in HT-29 cells showed by regulating the ERK1/2-mediated NF-κB pathway. Results of this study suggest that H. discus hannai hemolymph is involved in the regulation of Gram-negative bacteria-related inflammatory immune responses in human colonic epithelial cells.Copyright © 2023. Published by Elsevier Ltd.