大豆蛋白被认为是具有一系列重要生物学功能的优质蛋白。然而,由于大豆蛋白的溶解性差和较高的致敏性,其应用受到限制。它的多肽一直是人们感兴趣的,因为它们发挥着与大豆蛋白相同的生物学功能,但更容易吸收,更稳定和可溶,并且具有较低的致敏性。此外,最近的研究发现,化学部分连接到肽可以改善它们的特性,包括它们的生物分布,药代动力学,和生物活性,毒性较低。因此,这项研究旨在获得科学证据,以支持与阿洛酮糖(OT-AL)或D-甘露糖(OT-Man)缀合的大豆寡肽(OT)的进一步应用和安全使用。抗炎,细胞毒性,和OT的遗传毒性,OT-AL,并对OT-Man进行了调查。结果表明,OT,AL,伙计,OT-AL,和OT-Man在高达1000微克/毫升的剂量是无毒的HepG2(肝癌细胞),HEK293(肾细胞),LX-2(肝星状细胞),和成熟前-3T3-L1(成纤维细胞和脂肪细胞,分别),而在高剂量下略微延迟RAW264.7细胞(巨噬细胞)的增殖。此外,高达800µg/mL的寡肽对分离的人外周血单个核细胞(PBMC)没有毒性,也不会诱导人红细胞(RBC)溶血.OT-Man(200和400µg/mL),但不是OT,AL,伙计,OT-AL,显著降低脂多糖(LPS)刺激RAW264.7细胞NO的产生和诱导型一氧化氮合酶(iNOS)和环氧合酶-2(COX2)的表达,表明大豆肽的甘露糖缀合对LPS刺激的炎症具有抑制作用。此外,OT-AL(200和400µg/mL)和OT-Man(400µg/mL)显着降低了LPS刺激的白介素6(IL-6)的分泌。肿瘤坏死因子-α(TNF-α)水平显着降低OT(400µg/mL),AL(400µg/mL),OT-AL(200µg/mL),和OT-Man(200和400µg/mL)在LPS刺激的细胞中。肽与AL或Man的缀合可能增强抑制细胞因子分泌的抗炎能力。由于OT-Man表现出抑制LPS诱导的巨噬细胞炎症的高潜力,然后在细菌和果蝇中评估其致突变性能力。这些发现表明OT-Man不会触发DNA突变,并且是基因组安全的。这项研究为共轭大豆肽的健康优势和安全使用提供了可能的见解。
Soy protein is considered to be a high-quality protein with a range of important biological functions. However, the applications of soy protein are limited due to its poor solubility and high level of allergenicity. Its peptides have been of interest because they exert the same biological functions as soy protein, but are easier to absorb, more stable and soluble, and have a lower allergenicity. Moreover, recent research found that an attachment of chemical moieties to peptides could improve their properties including their biodistribution, pharmacokinetic, and biological activities with lower toxicity. This study therefore aimed to acquire scientific evidence to support the further application and safe use of the soybean oligopeptide (OT) conjugated with allulose (OT-AL) or D-mannose (OT-Man). The anti-inflammation, cytotoxicity, and genotoxicity of OT, OT-AL, and OT-Man were investigated. The results showed that OT, AL, Man, OT-AL, and OT-Man at doses of up to 1000 µg/mL were not toxic to HepG2 (liver cancer cells), HEK293 (kidney cells), LX-2 (hepatic stellate cells), and pre- and mature-3T3-L1 (fibroblasts and adipocytes, respectively), while slightly delaying the proliferation of RAW 264.7 cells (macrophages) at high doses. In addition, the oligopeptides at up to 800 µg/mL were not toxic to isolated human peripheral blood mononuclear cells (PBMCs) and did not induce hemolysis in human red blood cells (RBCs). OT-Man (200 and 400 µg/mL), but not OT, AL, Man, and OT-AL, significantly reduced the production of NO and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2) stimulated by lipopolysaccharide (LPS) in RAW 264.7 cells, suggesting that the mannose conjugation of soy peptide had an inhibitory effect against LPS-stimulated inflammation. In addition, the secretion of interleukin-6 (IL-6) stimulated by LPS was significantly reduced by OT-AL (200 and 400 µg/mL) and OT-Man (400 µg/mL). The tumor necrosis factor-α (TNF-α) level was significantly decreased by OT (400 µg/mL), AL (400 µg/mL), OT-AL (200 µg/mL), and OT-Man (200 and 400 µg/mL) in the LPS-stimulated cells. The conjugation of the peptides with either AL or Man is likely to be enhance the anti-inflammation ability to inhibit the secretion of cytokines. As OT-Man exhibited a high potential to inhibit LPS-induced inflammation in macrophages, its
mutagenicity ability was then assessed in bacteria and Drosophila. These findings showed that OT-Man did not trigger DNA mutations and was genome-safe. This study provides possible insights into the health advantages and safe use of conjugated soybean peptides.