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从水稻衍生肽中分离血红素加氧酶-1诱导剂。

Isolation of the hemeoxygenase-1 inducer from rice-derived peptide.

作者信息

Kawakami Kayoko, Moritani Chie, Hatanaka Tadashi, Tsuboi Seiji

机构信息

School of Pharmacy, Shujitsu University, 1-6-1 Nishigawara, Naka-ku, Okayama 703-8516, Japan.

Okayama Prefectural Technology Center for Agriculture, Forestry and Fisheries, Research Institute for Biological Sciences (RIBS), 7549-1 Kibichuo-cho, Kaga-‍gun, Okayama 716-1241, Japan.

出版信息

J Clin Biochem Nutr. 2022 Jul;71(1):41-47. doi: 10.3164/jcbn.21-125. Epub 2022 Jan 25.

DOI:10.3164/jcbn.21-125
PMID:35903607
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9309089/
Abstract

Bioactive peptides with various health benefits have been reported from rice protein hydrolysates. We previously showed that rice-derived peptides (RP) increased intracellular glutathione levels and induced the expression of γ-glutamylcysteine synthetase, which is regulated by nuclear transcription factor-erythroid 2-related factor 2 (Nrf2). Heme oxygenase-1 (HO-1) is an important Nrf2 downstream antioxidant enzyme that protects against oxidative stress. This study aimed to investigate the protective effects of RP on hydrogen peroxide (HO)-induced oxidative stress in human hepatoblastoma cell line HepG2 and identified HO-1 induced peptides from RP. Pretreatment of cells with RP reduced the cytotoxicity caused by HO in a dose-dependent manner. Moreover, RP induced HO-1 expression in a concentration- and time-dependent manner. Next, we attempted to isolate the HO-1 inducer from RP by bioactivity-guided fractionation. Purification of the active peptides using a Sep-Pak C18 cartridge and reversed-phase HPLC, followed by sequence analysis by mass spectrometry, led to the identification of the three peptides. These peptides effectively reduced HO-induced oxidative stress. Among them, only P3 (peptide sequence: RSAVLLSH) increased HO-1 protein expression. Additionally, the knockdown of Nrf2 suppressed the induction of HO-1 expression by P3. Our results indicated that P3 identified from RP induced HO-1 by activating the Nrf2 signaling pathway.

摘要

已报道大米蛋白水解物中存在具有多种健康益处的生物活性肽。我们之前的研究表明，大米衍生肽（RP）可提高细胞内谷胱甘肽水平，并诱导γ-谷氨酰半胱氨酸合成酶的表达，该酶受核转录因子红细胞2相关因子2（Nrf2）调控。血红素加氧酶-1（HO-1）是一种重要的Nrf2下游抗氧化酶，可抵御氧化应激。本研究旨在探讨RP对人肝癌细胞系HepG2中过氧化氢（HO）诱导的氧化应激的保护作用，并从RP中鉴定出诱导HO-1的肽。用RP预处理细胞可剂量依赖性地降低HO引起的细胞毒性。此外，RP以浓度和时间依赖性方式诱导HO-1表达。接下来，我们尝试通过生物活性导向分级分离从RP中分离出HO-1诱导剂。使用Sep-Pak C18柱和反相高效液相色谱法纯化活性肽，随后通过质谱进行序列分析，从而鉴定出三种肽。这些肽有效地降低了HO诱导的氧化应激。其中，只有P3（肽序列：RSAVLLSH）增加了HO-1蛋白表达。此外，敲低Nrf2可抑制P3对HO-1表达的诱导。我们的结果表明，从RP中鉴定出的P3通过激活Nrf2信号通路诱导HO-1。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/a39d3305b946/jcbn21-125f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/15400ede3442/jcbn21-125f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/c24afdb16aa4/jcbn21-125f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/6e53a03af76e/jcbn21-125f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/ccac375978b2/jcbn21-125f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/8f7e3e698685/jcbn21-125f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/a39d3305b946/jcbn21-125f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/15400ede3442/jcbn21-125f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/c24afdb16aa4/jcbn21-125f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/6e53a03af76e/jcbn21-125f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/ccac375978b2/jcbn21-125f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/8f7e3e698685/jcbn21-125f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8b/9309089/a39d3305b946/jcbn21-125f06.jpg

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