基于体外模拟消化与蛋白质组学技术的鸡内金研究与分析

Digestion-promoting Function and Protein Composition of Galli Gigerii Endothelium Corneum:A Study Based on in Vitro Simulated Digestion and Proteomics

目的:通过体外模拟消化研究与蛋白质组学技术,分析鸡内金的促消化作用及其蛋白质组成,为完善鸡内金质量评价体系提供参考。方法:采集市售鸡内金,鉴定其性别来源、测量其厚度、记录其颜色;利用静态多室多酶体外消化模型,考察不同性状鸡内金对底物消化率的影响,并利用蛋白组学技术鉴定鸡内金中蛋白质组分。结果:添加鸡内金后,底物干物质消化率与能量消化率分别增加了5.8%和8.2%;鸡内金颜色及原动物性别对底物干物质消化率与能量消化率均无显著影响;厚度与干物质消化率相关性显著(P<0.05),但未遵循线性规律。鸡内金厚度与淀粉酶活力呈极显著正相关(P<0.01),随厚度增加,淀粉酶活力上升。共从鸡内金中鉴定到1541种蛋白质,相对丰度较高的蛋白有胃动蛋白2、富含半胱氨酸的分泌蛋白、埃兹蛋白等。所测蛋白质主要富集在代谢、核糖体、碳代谢、蛋白酶体、三羧酸循环等京都基因与基因组百科全书(KEGG)通路上,其中富集到代谢通路上的蛋白质最多(240种)。结论:体外模拟消化实验在一定程度上可体现鸡内金促消化能力,可为鸡内金或其他消食类中药质量评价提供参考;推测胃动蛋白2和埃兹蛋白与鸡内金健脾消食的功效有关。

Objective:To study the digestion-promoting function and protein composition of Galli Gigerii Endothelium Corneum(GGEC)via simulated digestion in vitro and proteomics tools and provide reference for improving the quality evaluation system of GGEC.Methods:The GGEC sample was purchased from the market,with the animal sex identified,the thickness measured,and the color recorded.A static multi-compartment and multi-enzyme digestion model was used to investigate the effects of the GGEC samples with different animal sexes,thickness,and colors on substrate digestibility.Proteomic tools were employed to identify the proteins in GGEC.Results:The addition of GGEC increased the dry matter digestibility and energy digestibility of the substrate by 5.8%and 8.2%,respectively.The color and animal sex of GGEC had no significant effect on the dry matter digestibility or energy digestibility of the substrate.The thickness of GGEC was correlated with dry matter digestibility(P<0.05),which did not follow a linear rule.GGEC thickness had a positive correlation with amylase activity(P<0.01).In addition,a total of 1541 proteins were identified from GGEC,of which gastrokine 2,cysteine-rich secretory protein,ezrin,etc.had high relative abundance.The identified proteins were mainly enriched in the metabolism,ribosome,carbon metabolism,proteasome,and tricarboxylic acid cycle pathways,and the metabolism pathway was enriched with the most proteins(240).Conclusion:The simulated digestion in vitro can reflect the digestion-promoting ability of GGEC and provide reference for the quality evaluation of GGEC or other Chinese medicines for digestion.Gastrokine 2 and ezrin may be associated with the role of GGEC in invigorating spleen and promoting digestion.