乳清蛋白浓缩物及其水解物对大鼠嗜碱性白血病(RBL)-2H3细胞过敏参数的比较

Q2 Agricultural and Biological Sciences Korean Journal for Food Science of Animal Resources Pub Date : 2018-09-01 Epub Date: 2018-09-30 DOI:10.5851/kosfa.2018.e16
Hana Kim, Sung-Il Ahn, Jin-Woo Jhoo, Gur-Yoo Kim
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引用次数: 6

摘要

本研究比较了乳清蛋白浓缩物(WPC)和水解乳清蛋白的抗过敏作用。用胰蛋白酶和α-凝乳胰蛋白酶酶切制备WPC水解产物,酶切8 h后冷冻干燥。采用实时聚合酶链式反应(real - time polymerase chain reaction, PCR)测定大鼠嗜碱性白血病(RBL)-2H3细胞的过敏参数为β-己糖氨酸酶的脱粒和释放、肿瘤坏死因子(tumor necrosis factor, TNF)-α的释放以及IL-1β、IL-4和IL-10的表达变化。WPC水解产物制备过程中,从0 ~ 10 min水解迅速增加,从1 h开始逐渐缓慢增加,最终水解度为78.50%。SDS-PAGE分析显示,与WPC相比,WPC水解物中几个蛋白带的强度降低。与WPC处理相比,ige诱导的RBL-2H3细胞β-己糖氨酸酶释放量明显降低。W500(500µg/mL)对β-己糖氨酸酶释放的抑制作用最小,但W500与W1000(1000µg/mL)之间无显著差异(p
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Comparison of Allergic Parameters between Whey Protein Concentrate and Its Hydrolysate in Rat Basophilic Leukemia (RBL)-2H3 Cells.

This study was conducted to compare the anti-allergic effects of a whey protein concentrate (WPC) and WPC hydrolysate. WPC hydrolysate was prepared using enzymatic digestion for 8 h with trypsin and α-chymotrypsin, after which it was freeze-dried. The allergic parameters assessed in rat basophilic leukemia (RBL)-2H3 cells were degranulation and release of β-hexosaminidase, release of tumor necrosis factor (TNF)-α, and changes in the expression of IL-1β, IL-4, and IL-10 by real time polymerase chain reaction (PCR). During preparation of the WPC hydrolysate, hydrolysis increased rapidly from 0 to 10 min and then gradually increased slowly from 1 h onwards, achieving a final degree of hydrolysis of 78.50%. The SDS-PAGE analysis revealed a reduction in the intensity of several protein bands in the WPC hydrolysate compared to the WPC. IgE-induced β-hexosaminidase release from RBL-2H3 cells was decreased to a higher degree following treatment with the hydrolysate compared to WPC treatment. W500 (500 µg/mL WPC) showed the least inhibition of β-hexosaminidase release, but there was no significant difference between W500 and W1000 (1,000 µg/mL) (p<0.05). H1000 (1,000 µg/mL WPC hydrolysate) inhibited β-hexosaminidase release by 39%. Compared to the control, treatment with H1000 decreased TNF-α secretion to 11.87 pg/mL. The gene expression levels of IL-1β, IL-4, and IL-13 were all significantly decreased in hydrolysate (p<0.05). In the case of IL-1β and IL-4, the expression levels in W1000 treated cells were decreased by 73.67% and 65%, respectively, and that of IL-13 was decreased by 66.43% compared to the control.

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CiteScore
1.22
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审稿时长
4-8 weeks
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