Anti-inflammatory action of two novel peptides derived from peanut worms (Sipunculus nudus) in lipopolysaccharide-induced RAW264.7 macrophages.,Food & Function

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Anti-inflammatory action of two novel peptides derived from peanut worms (Sipunculus nudus) in lipopolysaccharide-induced RAW264.7 macrophages.
Food & Function ( IF 5.1 ) Pub Date : 2019-12-18 , DOI: 10.1039/c9fo02178g
Papassara Sangtanoo ₁ , Piroonporn Srimongkol , Tanatorn Saisavoey , Onrapak Reamtong , Aphichart Karnchanatat

Affiliation

Peanut worm (Sipunculus nudus Linn.) protein was hydrolyzed by three proteases, and NO scavenging activity of the protein hydrolysates was evaluated. The hydrolysate obtained using Alcalase® showed the highest NO scavenging activity. This hydrolysate was fractionated using 10-, 5-, and 3 kDa molecular weight cut-off membranes, and the lowest MW fraction (<3 kDa) exhibited the highest NO scavenging activity. The <3 kDa fraction was further purified by gel filtration and high-performance liquid chromatographies. The peptides in the HPLC fraction with the strongest anti-NO activity were identified by quadrupole-time-of-flight mass spectrometry as LSPLLAAH (821.48 Da) and TVNLAYY (843.42 Da). Both peptides and the corresponding pure synthetic peptides inhibited NO production by RAW 264.7 macrophages without cytotoxicity. Quantitative real-time RT-PCR analysis showed that peptides LSPLLAAH and TVNLAYY reduced expression of proinflammatory cytokine genes iNOS, IL-6, TNF-α, and COX-2 in RAW 264.7 macrophages, suggesting that these peptides are novel anti-inflammatory candidates.

中文翻译：

花生蠕虫（Sipunculus nudus）的两种新型肽在脂多糖诱导的RAW264.7巨噬细胞中的抗炎作用。

用三种蛋白酶水解花生蠕虫（Sipunculus nudus Linn。）蛋白，并评估该蛋白水解产物的NO清除活性。使用Alcalase®获得的水解产物显示出最高的NO清除活性。使用10、5和3 kDa分子量截留膜对水解产物进行分馏，最低的MW馏分（<3 kDa）表现出最高的NO清除活性。通过凝胶过滤和高效液相色谱法进一步纯化<3 kDa的馏分。通过四极杆飞行时间质谱法鉴定出HPLC级分中具有最强抗NO活性的肽为LSPLLAAH（821.48 Da）和TVNLAYY（843.42 Da）。肽和相应的纯合成肽均可抑制RAW 264.7巨噬细胞产生NO，而无细胞毒性。

更新日期：2020-02-10

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