Shimizu Miho, Sugai Wataru, Ohto-Fujita Eri, Atomi Aya, Nogawa Norio, Takamiya Koichi, Yoshinaga Hisao, Asano Yoshihide, Yamashita Takashi, Sato Shinichi, Enomoto Atsushi, Hatakeyama Nozomi, Yasuda Shunsuke, Tanaka Kazuya, Atomi Tomoaki, Harada Kenji, Hasebe Yukio, Watanabe Toshiyuki, Atomi Yoriko
Material Health Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, Tokyo 184-8588, Japan.
Division of Applied Chemistry, Graduate School of Engineering, Tokyo University of Agriculture and Technology, Tokyo 184-8588, Japan.
Int J Mol Sci. 2025 Sep 18;26(18):9102. doi: 10.3390/ijms26189102.
Eggshell membrane (ESM) is composed of approximately 90% protein. Our previous studies in healthy adults demonstrated that two months of daily ESM intake improved respiratory function, zigzag walking speed, and skin elasticity. The present study aims to address the knowledge gap regarding the in vivo effects of ESM in the context of inflammatory bowel disease (IBD). Proteomic analysis was performed on powdered ESM used as a dietary supplement. To investigate its pharmacokinetics in mice, tritium (H)-labeled ESM was prepared using the Li(n,α)H nuclear reaction. The therapeutic potential of ESM was further examined in a 2.0% dextran sulfate sodium (DSS)-induced murine model of IBD. In addition, fecal samples from both mice and healthy human subjects were analyzed using a modified terminal restriction fragment length polymorphism (T-RFLP) method. Lysozyme C (LYZ) was the most abundant protein (47%), followed by lysyl oxidase (12%) in ESM used in this study. H-ESM was mixed with MediGel, and orally administered to mice. Radioactivity levels were measured in blood, organs (duodenum, small intestine, large intestine, liver, kidney, lung, skin), and rectal feces at 0.5, 2, 5, 24, 48, and 72 h post-administration. Radioactivity in feces indicated excretion of undigested components, while systemic distribution suggested potential whole-body effects of ESM. Oral ESM and LYZ significantly alleviated body weight loss, diarrhea, and hematochezia in a DSS-induced murine model of IBD, leading to a significantly lower disease activity index on day 3 and showing a similar trend on day 5. Gut microbiota analysis showed increased in the DSS group, while the ESM + DSS group maintained levels similar to the control. In humans, a double-blind, randomized controlled trial was conducted to evaluate the effects of ESM on gut microbiota in healthy adults. Participants received either ESM or placebo for 8 weeks. revealed a significant increase in alpha diversity at weeks 1 and 8 in the ESM group ( < 0.05), with between-group differences evident from week 1 ( < 0.01). ESM intake reduced and significantly increased and at weeks 4 and 8. These findings suggest ESM supplementation promotes beneficial modulation of gut microbiota. These findings suggest that ESM, through its major protein components such as LYZ, may serve as a promising dietary intervention for maintaining intestinal health and mitigating inflammation in the context of IBD.
蛋壳膜（ESM）约由90%的蛋白质组成。我们之前对健康成年人的研究表明，连续两个月每日摄入ESM可改善呼吸功能、之字形行走速度和皮肤弹性。本研究旨在填补关于ESM在炎症性肠病（IBD）背景下的体内作用的知识空白。对用作膳食补充剂的粉状ESM进行了蛋白质组学分析。为了研究其在小鼠体内的药代动力学，利用锂（n，α）氢核反应制备了氚（H）标记的ESM。在2.0%硫酸葡聚糖钠（DSS）诱导的IBD小鼠模型中进一步研究了ESM的治疗潜力。此外，使用改良的末端限制性片段长度多态性（T-RFLP）方法分析了小鼠和健康人类受试者的粪便样本。溶菌酶C（LYZ）是本研究中使用的ESM中含量最丰富的蛋白质（占47%），其次是赖氨酰氧化酶（占12%）。将H-ESM与MediGel混合后口服给予小鼠。在给药后0.5、2、5、24、48和72小时测量血液、器官（十二指肠、小肠、大肠、肝脏、肾脏、肺、皮肤）和直肠粪便中的放射性水平。粪便中的放射性表明未消化成分的排泄，而全身分布表明ESM可能具有全身作用。口服ESM和LYZ可显著减轻DSS诱导的IBD小鼠模型中的体重减轻、腹泻和便血，在第3天导致疾病活动指数显著降低，并在第5天呈现类似趋势。肠道微生物群分析显示DSS组的[此处原文缺失相关内容]增加，而ESM+DSS组维持与对照组相似的水平。在人类中，进行了一项双盲、随机对照试验，以评估ESM对健康成年人肠道微生物群的影响。参与者接受ESM或安慰剂治疗8周。结果显示，ESM组在第1周和第8周时α多样性显著增加（P<0.05），从第1周起组间差异明显（P<0.01）。在第4周和第8周时，摄入ESM可减少[此处原文缺失相关内容]并显著增加[此处原文缺失相关内容]和[此处原文缺失相关内容]。这些发现表明补充ESM可促进肠道微生物群的有益调节。这些发现表明，ESM通过其主要蛋白质成分如LYZ，可能成为在IBD背景下维持肠道健康和减轻炎症的一种有前景的饮食干预措施。
