Article(id=1320881414761992986, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320881182762447607, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025070351, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1753804800000, receivedDateStr=2025-07-30, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792852270068, onlineDateStr=2026-10-24, pubDate=1786723200000, pubDateStr=2026-08-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792852270068, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792852270068, creator=13701087609, updateTime=1792852270068, updator=13701087609, issue=Issue{id=1320881182762447607, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='16', pageStart='1', pageEnd='537', issueExtLink='null', onlineDate='null', pubDate='1786723200000', pubDateStr='2026-08-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792852214756, creator='13701087609', updateTime=1792852214756, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=461, endPage=470, ext={EN=ArticleExt(id=1320881414980096795, articleId=1320881414761992986, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=In Vitro Simulated Digestive Properties of Walnut Pellicle Polyphenols and Their Antioxidant and Hypoglycemic Activities, columnId=1261343846965830077, journalTitle=Science and Technology of Food Industry, columnName=Nutrition and Healthcare, runingTitle=null, highlight=null, articleAbstract=

To investigate the effects of in vitro digestion on the phenolic compounds and bioactivity of walnut pellicle polyphenols extracts, the walnut pellicle polyphenols extracts were subjected sequentially to simulated oral, gastric and small intestinal digestion in this study. The total phenolic content of the walnut pellicle polyphenols extracts was measured at different stages of digestion. Antioxidant and hypoglycemic activities of the walnut pellicle polyphenols extracts were evaluated using DPPH radical scavenging rates, ABTS+ radical scavenging rates, and α-glucosidase and α-amylase inhibition rates. The results showed that digestion led to a significant decrease (P<0.05) in the total phenolic content and bioaccessibility, and the total phenolic content decreased from 695.27 mg GAE/g to 437.58 mg GAE/g, with the most significant degradation in the small intestine stage. A total of 224 phenolics were identified in the walnut pellicle polyphenols, and the main polyphenols were gallic acid, corilagin, and chlorogenic acid. During digestion, the antioxidant capacity of walnut pellicle polyphenols decreased (P<0.05), with DPPH· radical scavenging rate dropping from 90.18% to 35.73% and ABTS+ radical scavenging rate declining from 90.84% to 47.33%. Simultaneously, their hypoglycemic activity also weakened (P<0.05), manifested by a 36.1% decrease in α-glucosidase inhibition rate and a 72.44% decrease in α-amylase inhibition rate. At the end of digestion, the walnut pellicle polyphenols were degraded or transformed, but they still exerted some biological activities. During the digestion process, the changes of DPPH∙ and ABTS+∙ scavenging rates, α-glucosidase and α-amylase inhibition rates were positively correlated with the changes of total phenol content. This study demonstrated that walnut pellicle polyphenols had the potential to develop functional foods, and also provided a theoretical basis for the resource utilization of walnut processing by-products.

, authors=Ying CHEN1, Yizhen QIN1, Yanling LU1, Guohui YUAN1, Jiaojiao YANG1, Long HAN1, 2, 3, *, Fangyu FAN1, 2, 3, *, authorsList=Ying CHEN, Yizhen QIN, Yanling LU, Guohui YUAN, Jiaojiao YANG, Long HAN, Fangyu FAN, authorCompany=null, correspAuthors=Long HAN, Fangyu FAN, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright © 2026 Science and Technology of Food Industry. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1320881416351634214, articleId=1320881414761992986, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=核桃内种皮多酚的体外模拟消化特性及其抗氧化、降血糖活性, columnId=1261343850979779009, journalTitle=食品工业科技, columnName=营养与保健, runingTitle=null, highlight=null, articleAbstract=

为研究核桃内种皮多酚提取物在体外模拟消化过程中的酚类物质和生物活性的变化,本文对核桃内种皮多酚提取物依次进行模拟口腔消化、胃消化和小肠消化,测定核桃内种皮多酚提取物在不同消化阶段的总酚含量,以DPPH·、ABTS+·清除率、α-葡萄糖苷酶抑制率、α-淀粉酶抑制率为指标,考察核桃内种皮多酚提取物抗氧化、降血糖活性的变化情况。结果表明,消化导致总酚含量显著降低(P<0.05),总酚含量由695.27 mg GAE/g降至437.58 mg GAE/g,在小肠阶段的降解最为显著。核桃内种皮多酚共鉴定出224种酚类物质,主要酚类物质有没食子酸、柯里拉京、绿原酸等。在消化过程中,核桃内种皮多酚的抗氧化能力显著下降(P<0.05),DPPH·清除率从90.18%降至35.73%,ABTS+·清除率从90.84%降至47.33%。同时,其降血糖活性也显著降低(P<0.05),表现为α-葡萄糖苷酶抑制率下降36.18%,α-淀粉酶抑制率下降72.44%。消化结束后,核桃内种皮多酚发生降解或转变,但仍能发挥一定的生物活性。在消化过程中,DPPH∙和ABTS+∙清除率、α-葡萄糖苷酶和α-淀粉酶抑制率的变化与总酚含量变化呈正相关。本研究表明核桃内种皮多酚具备开发功能性食品的潜力,为核桃加工副产物的资源化利用提供了理论依据。

, authors=陈颖1, 秦毅臻1, 鲁燕玲1, 袁国惠1, 杨娇娇1, 韩龙1, 2, 3, *, 范方宇1, 2, 3, *, authorsList=陈颖, 秦毅臻, 鲁燕玲, 袁国惠, 杨娇娇, 韩龙, 范方宇, authorCompany=null, correspAuthors=韩龙, 范方宇, authorNote=

陈颖(2002−),女,硕士研究生,研究方向:食品加工与安全,E-mail:

, correspAuthorsNote=
韩龙(1987−),男,博士,助理研究员,研究方向:食品微生物生态、发酵微生物、食品加工副产物与农林废弃物资源化利用,E-mail:。
范方宇(1979−),男,博士,教授,研究方向:农林产品加工,E-mail:。
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2Forest Resources Exploitation and Utilization Engineering Research Center for Grand Health of Yunnan Provincial Universities, 650224, China
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Journal of Shanxi Agricultural University (Natural Science Edition), 2022, 42(3): 98−105.], articleTitle=null, refAbstract=null), Reference(id=1320881709797725150, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=52, rfOrder=66, authorNames=null, journalName=null, refType=null, unstructuredReference=LI H Q, CAO Y X, HUANG F J. Comparative study of DPPH∙ and ABTS+∙ methods on the antioxidant synergistic effects of polyphenols, flavonoids and polysaccharides of Dracocephalum rupestre Hance[J]. Journal of Shanxi Agricultural University (Natural Science Edition), 2022, 42(3): 98−105., articleTitle=null, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1320881692554941267, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, xref=1, ext=[AuthorCompanyExt(id=1320881692563329876, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, companyId=1320881692554941267, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1College of Biological and Food Engineering, Southwest Forestry University, Kunming 650224, China), AuthorCompanyExt(id=1320881692571718485, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, companyId=1320881692554941267, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1西南林业大学生物与食品工程学院,云南昆明 650224)]), AuthorCompany(id=1320881692659798870, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, xref=2, ext=[AuthorCompanyExt(id=1320881692668187479, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, companyId=1320881692659798870, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2Forest Resources Exploitation and Utilization Engineering Research Center for Grand Health of Yunnan Provincial Universities, 650224, China), AuthorCompanyExt(id=1320881692676576088, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, companyId=1320881692659798870, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2云南省高校大健康类森林资源开发利用工程研究中心,云南昆明 650224)]), AuthorCompany(id=1320881692764656473, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, xref=3, ext=[AuthorCompanyExt(id=1320881692773045082, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, companyId=1320881692764656473, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3Key Laboratory of Forest Disaster Warning and Control of Yunnan Province, Kunming 650224, China), AuthorCompanyExt(id=1320881692781433691, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, companyId=1320881692764656473, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3云南省森林灾害预警与控制实验室,云南昆明 650224)])], figs=[ArticleFig(id=1320881700184380301, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Fig.1, caption=Changes of total phenol content in walnut pellicle polyphenols through in vitro simulated digestion, figureFileSmall=7GQEf9ZbU5FoFlx1Zldz+g==, figureFileBig=F25jCeQcPmBkgWlWlnlhRg==, tableContent=null), ArticleFig(id=1320881700251489166, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=图1, caption=体外模拟消化中核桃内种皮多酚的总酚含量变化

注:不同小写字母表示不同消化阶段的核桃内种皮多酚之间存在显著差异(P<0.05)。

, figureFileSmall=7GQEf9ZbU5FoFlx1Zldz+g==, figureFileBig=F25jCeQcPmBkgWlWlnlhRg==, tableContent=null), ArticleFig(id=1320881700364735375, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Fig.2, caption=Classification of phenolic compounds of walnut pellicle polyphenols through in vitro simulated digestion, figureFileSmall=E+stVk+Eu9j16EaV7uuECQ==, figureFileBig=g3LnB0yqtUJeWRUfRueLDg==, tableContent=null), ArticleFig(id=1320881700431844240, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=图2, caption=体外模拟消化过程中核桃内种皮多酚的酚类化合物分类

注:A.未消化;B.口腔阶段;C.胃阶段;D.小肠阶段。

, figureFileSmall=E+stVk+Eu9j16EaV7uuECQ==, figureFileBig=g3LnB0yqtUJeWRUfRueLDg==, tableContent=null), ArticleFig(id=1320881700498953105, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Fig.3, caption=Changes in antioxidant ability of walnut pellicle polyphenols through in vitro simulated digestion, figureFileSmall=iRNhHe6BuBKpzfu/K6vo4w==, figureFileBig=r3ygq5N7Uqw2j1kHlZNIVw==, tableContent=null), ArticleFig(id=1320881700557673362, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=图3, caption=核桃内种皮多酚在体外模拟消化中抗氧化能力的变化

注:A. DPPH∙清除率;B. ABTS+∙清除率;不同小写字母表示不同消化阶段的核桃内种皮多酚之间存在显著差异(P<0.05);图4同。

, figureFileSmall=iRNhHe6BuBKpzfu/K6vo4w==, figureFileBig=r3ygq5N7Uqw2j1kHlZNIVw==, tableContent=null), ArticleFig(id=1320881700624782227, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Fig.4, caption=Changes in hypoglycemic activity of walnut pellicle polyphenols through in vitro simulated digestion, figureFileSmall=zt6VnS5YkpO1VxoD8wmGXQ==, figureFileBig=pbIrDsK+YdRKrlQw9/3F1A==, tableContent=null), ArticleFig(id=1320881700696085396, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=图4, caption=核桃内种皮多酚在体外模拟消化中降血糖活性的变化

注:A. α-葡萄糖苷酶抑制率;B. α-淀粉酶抑制率。

, figureFileSmall=zt6VnS5YkpO1VxoD8wmGXQ==, figureFileBig=pbIrDsK+YdRKrlQw9/3F1A==, tableContent=null), ArticleFig(id=1320881700792554389, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Fig.5, caption=Correlation analysis between in vitro activity and active ingredient release during walnut pellicle polyphenols digestion, figureFileSmall=DurU01hmL78uZ83q/nsb8w==, figureFileBig=EKfi5UzWSnM9DUx+Bh+5VA==, tableContent=null), ArticleFig(id=1320881700863857558, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=图5, caption=核桃内种皮多酚消化过程中体外活性与总酚含量之间的相关性分析

注:*表示相关性显著,P<0.05,**表示相关性极显著,P<0.01。

, figureFileSmall=DurU01hmL78uZ83q/nsb8w==, figureFileBig=EKfi5UzWSnM9DUx+Bh+5VA==, tableContent=null), ArticleFig(id=1320881700926772119, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Table 1, caption=

Major phenolic compounds of walnut pellicle polyphenols through in vitro simulated digestion

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物类别化合物名称相对丰度(×105)
未消化口腔胃小肠
酚酸类化合物没食子酸850.52±1.23d3370.79±39.20b1519.78±17.23c4400.64±8.18a
6-O-咖啡酰-D-葡萄糖651.86±1.94a118.12±2.18b21.34±0.33c17.89±0.11d
绿原酸13.97±0.05a3.73±2.77c8.66±0.05b6.30±0.01c
间羟基肉桂酸45.11±0.18b39.08±0.09c38.06±.016d93.71±0.18a
单宁类化合物柯里拉京2552.05±1.99a2301.10±17.92c1989.30±96.06d2465.06±62.90b
1-O-没食子酰基-D-葡萄糖596.92±5.29a236.57±4.63c280.79±7.66b143.16±4.61d
类黄酮类化合物金合欢素157.38±1.73a92.85±0.77b63.50±0.57c20.20±0.01d
香蜂草苷192.38±11.41a153.58±1.95b18.43±0.70d45.40±0.45c
6-羟基黄酮19.92±0.06c130.40±1.34b13.00±0.02d140.32±0.66a
圣草酚65.31±1.48b104.74±9.89a13.52±0.46d47.82±0.58c
槐黄烷酮 G118.35±2.62a82.17±0.23c89.71±0.004b73.18±0.36d
甲基紫檀素54.42±0.07b54.16±0.25b44.44±0.06c70.61±0.35a
芪类化合物白皮杉醇121.87±0.93a0.17±0.01c0.90±0.003b0.12±0.004d
其他酚类化合物2',4'-二羟基苯乙酮331.96±3.54a268.19±5.57b246.66±19.86b123.37±0.80c
2,6-二羟基苯乙酮255.64±2.47a241.21±1.92b79.94±0.04c61.16±0.05d
), ArticleFig(id=1320881701002269592, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=表1, caption=

体外模拟消化中核桃内种皮多酚的主要酚类化合物

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物类别化合物名称相对丰度(×105)
未消化口腔胃小肠
酚酸类化合物没食子酸850.52±1.23d3370.79±39.20b1519.78±17.23c4400.64±8.18a
6-O-咖啡酰-D-葡萄糖651.86±1.94a118.12±2.18b21.34±0.33c17.89±0.11d
绿原酸13.97±0.05a3.73±2.77c8.66±0.05b6.30±0.01c
间羟基肉桂酸45.11±0.18b39.08±0.09c38.06±.016d93.71±0.18a
单宁类化合物柯里拉京2552.05±1.99a2301.10±17.92c1989.30±96.06d2465.06±62.90b
1-O-没食子酰基-D-葡萄糖596.92±5.29a236.57±4.63c280.79±7.66b143.16±4.61d
类黄酮类化合物金合欢素157.38±1.73a92.85±0.77b63.50±0.57c20.20±0.01d
香蜂草苷192.38±11.41a153.58±1.95b18.43±0.70d45.40±0.45c
6-羟基黄酮19.92±0.06c130.40±1.34b13.00±0.02d140.32±0.66a
圣草酚65.31±1.48b104.74±9.89a13.52±0.46d47.82±0.58c
槐黄烷酮 G118.35±2.62a82.17±0.23c89.71±0.004b73.18±0.36d
甲基紫檀素54.42±0.07b54.16±0.25b44.44±0.06c70.61±0.35a
芪类化合物白皮杉醇121.87±0.93a0.17±0.01c0.90±0.003b0.12±0.004d
其他酚类化合物2',4'-二羟基苯乙酮331.96±3.54a268.19±5.57b246.66±19.86b123.37±0.80c
2,6-二羟基苯乙酮255.64±2.47a241.21±1.92b79.94±0.04c61.16±0.05d
), ArticleFig(id=1320881701073572761, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=EN, label=Table 2, caption=

Main hypoglycemic phenolics from walnut pellicle polyphenols during in vitro simulated digestion

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物名称相对丰度(×104)
未消化口腔胃小肠
高车前素269.15±1.49a104.72±4.64c148.67±0.29b16.49±0.03d
异绿原酸C(4,5)31.72±0.24a12.76±0.29c19.61±0.36b0.73±0.004d
槲皮素3-O-葡萄糖酸苷28.68±0.29a17.03±0.12c23.80±0.23b8.49±0.02d
草质素23.03±0.21a11.99±0.26c20.50±0.59b10.56±0.50d
香叶木素13.34±0.29a6.17±0.03c11.56±0.60b3.38±0.02d
金合欢素1573.76±17.27a928.50±7.71b634.98±5.73c202.03±0.08d
金圣草(黄)素441.39±2.06a242.75±0.60b120.12±0.56c88.61±0.35d
杨梅素301.09±0.93a239.07±1.16b139.94±0.10c67.15±0.04d
), ArticleFig(id=1320881701144875930, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320881414761992986, language=CN, label=表2, caption=

核桃内种皮多酚在体外模拟消化中主要发挥降血糖作用的酚类物质

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物名称相对丰度(×104)
未消化口腔胃小肠
高车前素269.15±1.49a104.72±4.64c148.67±0.29b16.49±0.03d
异绿原酸C(4,5)31.72±0.24a12.76±0.29c19.61±0.36b0.73±0.004d
槲皮素3-O-葡萄糖酸苷28.68±0.29a17.03±0.12c23.80±0.23b8.49±0.02d
草质素23.03±0.21a11.99±0.26c20.50±0.59b10.56±0.50d
香叶木素13.34±0.29a6.17±0.03c11.56±0.60b3.38±0.02d
金合欢素1573.76±17.27a928.50±7.71b634.98±5.73c202.03±0.08d
金圣草(黄)素441.39±2.06a242.75±0.60b120.12±0.56c88.61±0.35d
杨梅素301.09±0.93a239.07±1.16b139.94±0.10c67.15±0.04d
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核桃内种皮多酚的体外模拟消化特性及其抗氧化、降血糖活性
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陈颖 1 , 秦毅臻 1 , 鲁燕玲 1 , 袁国惠 1 , 杨娇娇 1 , 韩龙 1, 2, 3, * , 范方宇 1, 2, 3, *
食品工业科技 | 营养与保健 2026,47(16): 461-470
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食品工业科技 |营养与保健 2026 , 47 (16) : 461 -470
核桃内种皮多酚的体外模拟消化特性及其抗氧化、降血糖活性
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陈颖1 , 秦毅臻1, 鲁燕玲1, 袁国惠1, 杨娇娇1, 韩龙1, 2, 3, * , 范方宇1, 2, 3, *
作者信息
  • 1西南林业大学生物与食品工程学院,云南昆明 650224
  • 2云南省高校大健康类森林资源开发利用工程研究中心,云南昆明 650224
  • 3云南省森林灾害预警与控制实验室,云南昆明 650224
通讯作者:
韩龙(1987−),男,博士,助理研究员,研究方向:食品微生物生态、发酵微生物、食品加工副产物与农林废弃物资源化利用,E-mail:。
范方宇(1979−),男,博士,教授,研究方向:农林产品加工,E-mail:。
作者简介:

陈颖(2002−),女,硕士研究生,研究方向:食品加工与安全,E-mail:

In Vitro Simulated Digestive Properties of Walnut Pellicle Polyphenols and Their Antioxidant and Hypoglycemic Activities
Ying CHEN1 , Yizhen QIN1, Yanling LU1, Guohui YUAN1, Jiaojiao YANG1, Long HAN1, 2, 3, * , Fangyu FAN1, 2, 3, *
Affiliations
  • 1College of Biological and Food Engineering, Southwest Forestry University, Kunming 650224, China
  • 2Forest Resources Exploitation and Utilization Engineering Research Center for Grand Health of Yunnan Provincial Universities, 650224, China
  • 3Key Laboratory of Forest Disaster Warning and Control of Yunnan Province, Kunming 650224, China
出版时间: 2026-08-15 doi: 10.13386/j.issn1002-0306.2025070351
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为研究核桃内种皮多酚提取物在体外模拟消化过程中的酚类物质和生物活性的变化,本文对核桃内种皮多酚提取物依次进行模拟口腔消化、胃消化和小肠消化,测定核桃内种皮多酚提取物在不同消化阶段的总酚含量,以DPPH·、ABTS+·清除率、α-葡萄糖苷酶抑制率、α-淀粉酶抑制率为指标,考察核桃内种皮多酚提取物抗氧化、降血糖活性的变化情况。结果表明,消化导致总酚含量显著降低(P<0.05),总酚含量由695.27 mg GAE/g降至437.58 mg GAE/g,在小肠阶段的降解最为显著。核桃内种皮多酚共鉴定出224种酚类物质,主要酚类物质有没食子酸、柯里拉京、绿原酸等。在消化过程中,核桃内种皮多酚的抗氧化能力显著下降(P<0.05),DPPH·清除率从90.18%降至35.73%,ABTS+·清除率从90.84%降至47.33%。同时,其降血糖活性也显著降低(P<0.05),表现为α-葡萄糖苷酶抑制率下降36.18%,α-淀粉酶抑制率下降72.44%。消化结束后,核桃内种皮多酚发生降解或转变,但仍能发挥一定的生物活性。在消化过程中,DPPH∙和ABTS+∙清除率、α-葡萄糖苷酶和α-淀粉酶抑制率的变化与总酚含量变化呈正相关。本研究表明核桃内种皮多酚具备开发功能性食品的潜力,为核桃加工副产物的资源化利用提供了理论依据。

核桃内种皮  /  多酚  /  体外消化  /  抗氧化  /  降血糖

To investigate the effects of in vitro digestion on the phenolic compounds and bioactivity of walnut pellicle polyphenols extracts, the walnut pellicle polyphenols extracts were subjected sequentially to simulated oral, gastric and small intestinal digestion in this study. The total phenolic content of the walnut pellicle polyphenols extracts was measured at different stages of digestion. Antioxidant and hypoglycemic activities of the walnut pellicle polyphenols extracts were evaluated using DPPH radical scavenging rates, ABTS+ radical scavenging rates, and α-glucosidase and α-amylase inhibition rates. The results showed that digestion led to a significant decrease (P<0.05) in the total phenolic content and bioaccessibility, and the total phenolic content decreased from 695.27 mg GAE/g to 437.58 mg GAE/g, with the most significant degradation in the small intestine stage. A total of 224 phenolics were identified in the walnut pellicle polyphenols, and the main polyphenols were gallic acid, corilagin, and chlorogenic acid. During digestion, the antioxidant capacity of walnut pellicle polyphenols decreased (P<0.05), with DPPH· radical scavenging rate dropping from 90.18% to 35.73% and ABTS+ radical scavenging rate declining from 90.84% to 47.33%. Simultaneously, their hypoglycemic activity also weakened (P<0.05), manifested by a 36.1% decrease in α-glucosidase inhibition rate and a 72.44% decrease in α-amylase inhibition rate. At the end of digestion, the walnut pellicle polyphenols were degraded or transformed, but they still exerted some biological activities. During the digestion process, the changes of DPPH∙ and ABTS+∙ scavenging rates, α-glucosidase and α-amylase inhibition rates were positively correlated with the changes of total phenol content. This study demonstrated that walnut pellicle polyphenols had the potential to develop functional foods, and also provided a theoretical basis for the resource utilization of walnut processing by-products.

walnut pellicle polyphenols  /  polyphenols  /  in vitro digestion  /  antioxidant  /  hypoglycemia
陈颖, 秦毅臻, 鲁燕玲, 袁国惠, 杨娇娇, 韩龙, 范方宇. 核桃内种皮多酚的体外模拟消化特性及其抗氧化、降血糖活性. 食品工业科技, 2026 , 47 (16) : 461 -470 . DOI: 10.13386/j.issn1002-0306.2025070351
Ying CHEN, Yizhen QIN, Yanling LU, Guohui YUAN, Jiaojiao YANG, Long HAN, Fangyu FAN. In Vitro Simulated Digestive Properties of Walnut Pellicle Polyphenols and Their Antioxidant and Hypoglycemic Activities[J]. Science and Technology of Food Industry, 2026 , 47 (16) : 461 -470 . DOI: 10.13386/j.issn1002-0306.2025070351
核桃(Juglans regia L.)为胡桃科核桃属多年生乔木,是全球四大木本坚果树种之一,其兼具油料作物、经济林和生态修复性植物等多重属性,在农林产业中占重要地位[1−2]。核桃除传统可食用价值,其活性成分具有抗氧化、抗病毒、抗菌及神经保护等多种功能[3]。核桃内种皮是核桃仁表面的一层薄膜状结构,其质量仅占核桃仁的5%左右,集中了大量的酚类次生代谢产物,是核桃中生物活性成分的主要来源[4]。然而,核桃内种皮因含大量单宁类物质,有明显的食用苦涩味,在核桃仁深加工过程中常作为副产物被废弃,造成高价值生物活性物质的严重浪费[5]。因此,研究核桃种皮多酚的组成及生物活性,对实现农林加工副产物的高值化利用具有重要意义。
多酚,又名植物单宁,是具有多个羟基的芳香族化合物,广泛存在于水果、蔬菜和草本植物的根、皮、叶、果实和种子中[6],具有抗氧化、抑菌、抗炎[7]、降血糖[8−9]、调节肠道菌群[10]等多种活性,在学界受到广泛研究。然而,研究表明多酚的生理功能会受到人体消化的影响。He等[11]对22种果汁的生物活性物质进行体外模拟消化研究,发现消化后的多酚、多糖的含量及其抗氧化能力、α-葡萄糖苷酶和α-淀粉酶的抑制率显著提高,表明体外消化过程促进了活性成分的释放。Laya等[12]对木薯叶中的多酚进行体外模拟消化,发现消化后酚类物质含量下降,但其活性仍具较好的抗糖尿病和抗肥胖能力,表明体外消化会造成活性成分的降解或转化。因此,研究活性成分在消化过程中的变化具有重要意义。体外模拟消化实验是通过构建与动物体内相近的环境和消化酶体系从而高度还原体内消化过程,其具有成本低、周期短且重复性高等优势,是连接体外化学活性与体内生理功能的关键桥梁。
目前,关于核桃内种皮多酚的研究多关注于对其提取方法和生理功能的探索,对核桃内种皮多酚在体外模拟胃肠道的消化及活性变化的研究较少。基于此,本研究采用超声辅助乙醇法提取核桃内种皮多酚,经大孔吸附树脂纯化后探讨体外模拟消化特性,研究各消化阶段多酚含量、抗氧化能力和降血糖活性的变化,并通过UHPLC-MS/MS技术鉴定各消化阶段的酚类物质组成和相对含量,以期为核桃内种皮多酚在体内消化研究中提供理论参考,同时为其高值化利用提供新思路。
核桃 云南漾濞;氯化钠、磷酸氢二钠、磷酸二氢钾、无水碳酸钠 分析纯,广东光华科技股份有限公司;氢氧化钠、盐酸 分析纯,云南杨林工业开发区汕滇药业有限公司;粘蛋白、胃蛋白酶(猪胃粘膜)(3000 U/g)、胰蛋白酶(猪胰)(250 U/mg)、α-淀粉酶(5 U/mg)、α-葡萄糖苷酶(25 U/mg)、对硝基苯基-α-D-吡喃葡萄糖苷(p-NPG) 分析纯,上海源叶生物科技有限公司;1,1-二苯基-2-三硝基苯肼(DPPH)、2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS) HPLC≥98%,合肥巴斯夫生物科技有限公司。
FD5-3真空冷冻干燥机 北京华夏乐博仪器有限公司;JHF-500多功能粉碎机 南京贝帝实验仪器有限公司;KQ 5200 DE台式数控超声波清洗器 昆山市超声仪器有限公司;5804R高速中等通量台式离心机 德国艾本德公司;MD SpectraMax Plus 384酶标仪 美国伯腾公司;N-1100旋转真空蒸发仪 上海泉杰仪器有限公司;OTRAP 6500+质谱仪 美国应用生物系统公司;Nexera X2 LC-30AD超高压液相色谱仪 日本岛津公司。
参考许飞利等[13]方法,稍作修改。将干燥后的核桃内种皮粉碎,过60目筛,石油醚脱脂3次。以70%(v/v)乙醇作为提取剂,50 ℃超声(200 W,40 Hz)核桃内种皮55 min,料液比1:60(m:v)。4 ℃ 8000 r/min离心浸提液,沉淀按照相同程序进行3次浸提,混合上清液,50 ℃旋转蒸发直至除去乙醇。获得的核桃内种皮多酚提取物,冷冻干燥后储存在干燥器中,备用。参考Xie等[14]方法,稍作修改,进行样品的纯化。采用AB-8大孔吸附树脂对核桃内种皮粗多酚进行纯化,纯化后核桃内种皮多酚的纯度为69.53%,比纯化前(53.85%)提高了15.68%。
体外模拟消化模型包括口腔、胃和小肠消化。采用陈雅琪[15]的方法,稍作修改。口腔消化阶段:准确称取0.20 g样品,加入10.0 mL蒸馏水,与10.0 mL含有30.0 mg/mL粘蛋白的模拟唾液混合,混合物pH调至6.8,37 ℃恒温摇床(100 r/min)振荡10 min。胃消化阶段:经口腔消化的样品与20.0 mL的模拟胃液1:1(v:v)混合,调节pH至2.5,37 ℃恒温摇床(100 r/min)振荡2 h。小肠消化阶段:胃消化的样品pH调至7.0,依次添加2.0 mL模拟肠液、4.67 mL胆盐溶液、和3.33 mL胰酶溶液,37 ℃恒温摇床(100 r/min)振荡2 h,期间维持体系pH在7.0。将各阶段的模拟消化液在4 ℃、8000 r/min离心30 min,得到模拟口腔消化样品、胃消化样品,小肠消化样品,考察其总酚含量、抗氧化能力、α-葡萄糖苷酶抑制率和α-淀粉酶抑制率。
参考Xie等[14]方法,稍作修改。取0.5 mL样品,加入0.2 mol/L Folin试剂2.5 mL,7.5%(w/v)碳酸钠溶液2.0 mL,避光静置1 h,于765 nm处测量吸光度,以上操作重复3次。总没食子酸含量以没食子酸当量mg(GAE)/g核桃内种皮干重表示,没食子酸标准曲线y=7.6917x+0.0116,R2=0.9991。总酚含量按式(1)计算:
$\rm 总酚含量(\text{mg GAE/g})=\frac{C\times V\times N}{m} $
式中:C为由没食子酸标准曲线计算得出的样品中等效没食子酸浓度,mg/mL;V为溶液的体积,mL;N为稀释的倍数;m为样品的质量,g。
取1.0 mL各消化阶段样品冷冻干燥,加入80%(v/v)甲醇溶液1.0 mL,超声20 min后静置1 h,离心(4 ℃,8000 r/min),取上清液冷冻干燥。以200 μL 50%(v/v)甲醇溶液复溶样品,离心,取上清液于UHPLC-MS/MS检测。超高效液相色谱条件:HSS T3色谱柱进行分离,进样量10 μL,柱温40 ℃,流速200 µL/min。流动相体系:A相为0.1%(w/v)甲酸水溶液,B相为100%(v/v)乙腈。正离子模式液相梯度洗脱程序如下:0~2.5 min,0% B;2.5~9 min,0%~30% B;9~10 min,30%~100% B;10~15.4 min,100% B;15.4~15.5 min,100%~0% B;15.5~18 min,0% B。负离子模式液相梯度洗脱程序如下:0~2 min,0% B;2~2.5 min,0%~20% B;2~3.5 min,20%~40% B;3.5~4.5 min,40%~50% B;4.5~10 min,50%~100% B;10~15.4 min,100% B;15.4~15.5 min,100%~0% B;15.5~18 min,0% B,以上均为体积分数。样品经超高效液相色谱分离后用质谱仪进行质谱分析,使用加热电喷雾离子源进行离子化,质谱条件:电喷雾离子源温度为550 ℃;离子喷雾电压为5500 V(正离子模式)/−4500 V(负离子模式);离子源气体I、气体II和帘气分别设置为40、50和35 psi。采用多反应监测(Multiple reaction monitoring,MRM)模式检测待测离子对。
参考Ren等[16]的方法,稍作修改。配制0.1 mmol/L的DPPH溶液,避光储存。2.0 mL样品和等体积DPPH溶液混匀作为样品组(A1),室温避光反应30 min,517 nm处测定吸光值,以上操作重复3次。以2.0 mL样品和等体积无水乙醇混匀作为对照组(A2),以2.0 mL DPPH溶液和等体积无水乙醇混匀作为空白组(A0)。DPPH∙清除率按式(2)计算:
$\rm \text{DPPH}\cdot 清除率(\text{%})=\left(1-\frac{{A}_{1}-{A}_{2}}{{A}_{0}}\right)\times 100 $
式中:A1为样品吸光度;A2为对照组吸光度;A0为空白组吸光度。
参考Ren等[16]的方法,稍作修改。将ABTS溶液(7.0 mmo1/L)和过硫酸钾溶液(140.0 mmo1/L)1:1(v:v)混合,室温避光反应16 h,无水乙醇稀释至734 nm处吸光度为0.70±0.02的ABTS工作液。2.0 mL样品和等体积ABTS溶液混匀作为样品组(A1),以2.0 mL样品和等体积无水乙醇混匀作为对照组(A2),以2.0 mL ABTS溶液和等体积无水乙醇混匀作为空白组(A0)。室温避光反应6 min,734 nm处测定吸光值,以上操作重复3次。ABTS+∙清除率按式(3)计算:
$\rm {\text{ABTS}}^{{{}^+}}\cdot 清除率(\text{%})=\left(1-\frac{{A}_{1}-{A}_{2}}{{A}_{0}}\right)\times 100 $
式中:A1为样品吸光度;A2为对照组吸光度;A0为空白组吸光度。
α-葡萄糖苷酶抑制率的测定参考文献[17−18]的方法,稍作修改。将样品稀释至同一倍数,取50 μL转移到96孔板中,加入100 μL 1 U/mL α-葡萄糖苷酶溶液。37 ℃避光反应10 min,加入50 μL 0.005 mol/L p-NPG溶液,37 ℃反应20 min。加入50 μL 0.1 mol/L碳酸钠溶液终止反应,405 nm处测定其吸光值,以上操作重复3次。α-葡萄糖苷酶抑制率按式(4)计算:
$\rm \alpha\text{-}葡萄糖苷酶抑制率(\text{%})=\left(1-\frac{{A}_{1}-{A}_{2}}{{A}_{3}-{A}_{4}}\right)\times 100 $
式中:A1为含α-葡萄糖苷酶的提取物的吸光度;A2为不含α-葡萄糖苷酶(以pH6.8的磷酸盐缓冲液(Phosphate buffer saline,PBS)代替)的提取物的吸光度;A3为含α-葡萄糖苷酶的pH6.8的PBS的吸光度;A4为不含α-葡萄糖苷酶的pH6.8的PBS的吸光度。
参照Akhtar等[19]的方法,稍作修改。将样品稀释至同一倍数,取500 μL样品于2 mL离心管,加入500 μL α-淀粉酶于37 ℃水浴10 min,加入1.0% (w/v)的可溶性淀粉溶液500 μL继续水浴10 min,取出加入1.0 mL DNS显色剂,沸水浴5 min终止反应。蒸馏水定容至10.0 mL,冷却至室温,540 nm测吸光值,以上操作重复3次。α-淀粉酶抑制率按式(5)计算:
$\rm \alpha\text{-}淀粉酶抑制能力(\text{%})=\left(1-\frac{{A}_{1}-{A}_{2}}{{A}_{0}}\right)\times 100 $
式中:A1为含α-淀粉酶的提取物的吸光度;A2为不含α-葡萄糖苷酶(以pH6.9的PBS代替)的提取物的吸光度;A0为含α-淀粉酶的pH6.9的PBS的吸光度。
所有实验至少重复三次,结果以均数±标准差(X±SD)表示。采用IBM SPSS Statistics 26软件对数据进行统计学意义分析(显著性水平P<0.05表示有统计学意义),采用Origin 2021软件和ChiPlot免费在线平台(https://www.chiplot.online)绘图,相关性分析采用皮尔逊相关(Pearson correlation)系数法。
食品中酚类物质的释放受到人体消化过程的影响,其释放行为因样品类型而异[20]。多酚体外模拟消化实验,可高效、低成本地获取其在消化过程中的关键数据为后续研究和应用提供科学依据[21]。图1为体外模拟消化中核桃内种皮多酚的总酚含量变化,体外模拟消化中核桃内种皮总酚含量呈现下降趋势。未经消化的核桃内种皮总酚含量最高,达到695.27 mg GAE/g,是因为其含有大量的游离多酚[22];经口腔和胃消化后核桃内种皮总酚含量分别减少了34.81 mg GAE/g和87.46 mg GAE/g;小肠消化后,核桃内种皮总酚含量显著下降(P<0.05),为437.58 mg GAE/g。总酚含量在口腔消化后出现下降,是因为口腔消化作为消化过程的第一阶段唾液浸渍会对后续消化过程的多样性产生影响[23]。总酚含量在小肠消化后损失量是胃消化的2.95倍,说明与小肠环境相比,核桃内种皮多酚在胃的酸性环境中更加稳定。因为多酚在中性或微碱性pH与胰酶、胆汁盐存在的小肠液环境下易发生氧化降解或转化为新的化合物,导致核桃内种皮总酚含量降低;某些酚类物质(如槲皮素、没食子酸等)在酸性环境中相对稳定,因此胃消化阶段核桃内种皮总酚含量的下降较少[24]。Li等[25]在山药酚类化合物的体外模拟消化的研究中也有类似发现。综上,消化过程会使核桃内种皮总酚含量下降,但其不会完全降解。
基于UHPLC-MS/MS技术鉴定不同消化阶段核桃内种皮多酚的酚类化合物组成(图2)。核桃内种皮多酚由224种酚类化合物组成,其在口腔消化、胃消化和小肠消化阶段分别被检出224、225和220种酚类化合物。核桃内种皮多酚包括43种黄酮、40种酚酸、29种香豆素、20种黄烷酮、16种黄酮醇、11种芪类、10种查尔酮、10种异黄酮、7种木脂素、2种单宁、3种黄烷醇、25种其他酚类和8种其他黄酮类化合物;口腔消化阶段酚类化合物的组成与未消化阶段的一致,可能是因为口腔阶段的消化时间较短,酚类化合物没有足够的时间发生转化,但酚类化合物的相对含量仍有差异;与未消化阶段和口腔阶段相比,胃消化阶段增加了1种黄烷酮类化合物(芸香柚皮苷),黄烷酮类化合物的占比增加了0.40%,这可能是因为某些黄烷酮类化合物在胃的酸性环境中发生糖基化形成了黄烷酮苷[26],使黄烷酮类化合物的数量增加;在小肠消化阶段,黄酮醇类化合物的占比为6.82%,比胃消化阶段减少0.29%,木脂素类化合物、黄烷酮类化合物和其他黄酮类化合物也出现降解,是因为小肠的弱碱性环境不利于酚类化合物的稳定,酚类化合物经过小肠消化后发生较大程度的降解[26]。
基于上述对核桃内种皮多酚在体外模拟消化过程中酚类化合物的整体组成变化,选取各消化阶段样品中相对丰度较高的物质作为主要酚类化合物进行比较,分析其在体外模拟消化过程中的变化。并将主要酚类化合物按化学结构特征分为酚酸类化合物、单宁类化合物、类黄酮类化合物、芪类化合物和其他酚类化合物(表1)。
在酚酸类化合物中,没食子酸是核桃内种皮多酚的重要组成成分。其在未消化样品中具有较高丰度,为850.52×105±1.23×105,经体外消化后,其相对丰度在口腔阶段和小肠阶段显著增加(P<0.05),并于小肠阶段达到峰值(4400.64×105±8.18×105),这表明没食子酸在消化过程中可能被大量释放或由前体物质转化,且小肠是其释放的关键阶段。没食子酸是可水解单宁的降解产物,其在小肠阶段的相对丰度显著增加(P<0.05),可能与水解单宁在消化过程中的降解有关[27−28]。此外,没食子酸的相对丰度在胃消化阶段显著下降(P<0.05),原因可能是没食子酸可通过氢键、范德华力等非共价作用与胃蛋白酶结合形成复合物,导致其相对丰度下降[29]。6-O-咖啡酰-D-葡萄糖在消化过程中相对丰度整体呈下降趋势,相较于未消化阶段,其小肠消化阶段的相对丰度下降了97.26%。现已有研究关于胰酶对羧基连接的羟基肉桂酰葡萄糖酯具有特异性酯酶活性,6-O-阿魏酰-D-葡萄糖作为与其结构相似的底物被水解为游离阿魏酸和葡萄糖,因此,6-O-咖啡酰-D-葡萄糖也有可能被水解为咖啡酸和D-葡萄糖,导致其相对丰度显著下降(P<0.05)[30−31]。绿原酸在消化各阶段的相对丰度较低,且在口腔和小肠阶段均低于未消化样品,原因为绿原酸在口腔和小肠的碱性条件下水解生成3-O-咖啡酰奎宁酸和4-O-咖啡酰奎宁酸[32]。间羟基肉桂酸的相对丰度在消化过程中呈现先下降后上升的趋势。酚酸类化合物在消化过程中的变化可能体现酚类化合物的结合态向游离态转化的特征。
在单宁类化合物中,柯里拉京在未消化和消化各阶段具有较高的相对丰度(未消化:2552.05×105±1.99×105、口腔:2301.10×105±17.92×105、胃:1989.30×105±96.06×105、小肠:2465.06×105±62.90×105),说明其可能是核桃内种皮多酚中的重要成分。相较于未消化阶段,小肠消化结束后的柯里拉京和1-O-没食子酰基-D-葡萄糖的相对丰度分别下降了3.41%、76.02%,可能是因为柯里拉京和1-O-没食子酰基-D-葡萄糖作为水解单宁可被酸、碱和特定的酶催化水解,释放出鞣花酸或没食子酸等小分子酚酸[27−28],因此,单宁类化合物在消化后相对丰度降低,而其代谢物(没食子酸)相对丰度反而升高。
在类黄酮类化合物中,金合欢素、香蜂草苷、6-羟基黄酮、圣草酚、槲黄烷酮G和甲基紫檀素的相对丰度变化存在差异。其中,与未消化阶段相比,金合欢素、香蜂草苷和槐黄烷酮G的相对丰度在小肠消化阶段分别下降了87.16%、76.40%和38.17%。先前有研究表明环境的pH变化会影响消化过程中释放的类黄酮物质,当其暴露于中性或碱性环境时更容易分解或转化为分子量更小的酚酸[33−34]。6-羟基黄酮和圣草酚的相对丰度在口腔阶段升高,胃阶段下降,在小肠阶段再次升高,而甲基紫檀素的相对丰度在小肠阶段升高。
白皮杉醇为芪类化合物,在核桃内种皮多酚中相对丰度较高,但其相对丰度在口腔、胃和小肠消化阶段显著降低(P<0.05),可能是因为芪类化合物(如白藜芦醇)的化学结构不稳定,易受到消化道环境的影响而发生降解[35]。
其他酚类化合物包括2',4'-二羟基苯乙酮和2,6-二羟基苯乙酮,其相对丰度在小肠阶段相较于未消化时分别下降了62.84%和76.08%,表明其在消化后出现降解。
综上所述,核桃内种皮多酚的主要酚类化合物在体外模拟消化过程中的变化具有差异性,且体外模拟消化影响了核桃内种皮多酚中各类化合物的相对丰度与结构稳定性,其中小肠消化阶段是主要酚类化合物结构转化和释放的关键阶段。
通过不同的测定方法(DPPH、ABTS实验)评估核桃内种皮多酚在体外模拟消化过程中的抗氧化能力(图3),核桃内种皮多酚对自由基清除能力越大说明其抗氧化能力越强[36]。两种方法均表明,未经消化的核桃内种皮多酚的抗氧化能力最强(P<0.05),达到90%以上。随着消化过程的进行,DPPH∙和ABTS+∙清除率呈下降趋势,分别从90.18%和90.84%下降到35.73%和47.33%,与总酚含量变化的趋势一致。此外,核桃内种皮多酚在消化过程中对ABTS+∙清除效果均优于DPPH∙,是因为不同自由基的氧化作用机制存在差异[36]。消化结束后,核桃内种皮多酚发生降解或者转变,但仍具有较强的抗氧化能力。
在糖尿病治疗方案中,α-葡萄糖苷酶与α-淀粉酶因其在碳水化合物代谢中的核心作用被确立为关键药理学靶点,抑制α-葡萄糖苷酶和α-淀粉酶可有效控制餐后血浆葡萄糖水平,对于预防和管理糖尿病至关重要,因此,控制餐后高血糖的有效策略之一是使用α-葡萄糖苷酶和α-淀粉酶抑制剂延迟葡萄糖吸收[37]。核桃内种皮多酚对α-葡萄糖苷酶的抑制率如图4A所示。核桃内种皮多酚在消化后对α-葡萄糖苷酶的抑制率总体呈下降趋势,未消化的核桃内种皮多酚对α-葡萄糖苷酶的抑制率最高(P<0.05),达到98.05%;口腔消化后,其对α-葡萄糖苷酶的抑制率下降5.77%,胃消化阶段又上升至97.32%。未消化和经口腔、胃消化的核桃内种皮多酚对α-葡萄糖苷酶的抑制率无显著性差异(P>0.05),可能是由于核桃内种皮多酚中对酶活性具有抑制作用的酚类成分在口腔和胃消化阶段未被大量破坏或降解,仍能通过协同作用维持其抑制活性。小肠消化后,核桃内种皮多酚对α-葡萄糖苷酶的抑制率显著下降(P<0.05),为61.87%,可能是在肠消化的弱碱性环境下,黄酮类化合物的稳定性下降导致酚类物质含量的下降,因此其抑制α-葡萄糖苷酶的能力减弱[20]。相较于未消化阶段,小肠消化结束后核桃内种皮多酚对α-葡萄糖苷酶的抑制率下降了36.18%。核桃内种皮多酚对α-淀粉酶的抑制率(如图4B)所示,消化后其对α-淀粉酶的抑制率呈下降趋势,与总酚含量和抗氧化能力的变化趋势一致。未消化的核桃内种皮多酚对α-淀粉酶的抑制率达91.51%;口腔消化后,α-淀粉酶抑制率显著下降(P<0.05),为29.74%,可能核桃内种皮多酚在口腔消化阶段通过氢键和疏水作用与蛋白结合,形成不溶性复合物并发生沉淀,进而酶抑制作用减少[38];小肠消化后α-淀粉酶的抑制率为19.07%。相较于未消化阶段,小肠消化结束后核桃内种皮多酚对α-淀粉酶的抑制率下降了72.44%。
综上,核桃内种皮多酚体外模拟消化过程中,其对α-葡萄糖苷酶和α-淀粉酶的抑制率整体呈下降趋势。未消化样品对α-葡萄糖苷酶和α-淀粉酶均表现出较强抑制活性;其中,口腔和胃消化后α-葡萄糖苷酶抑制率仍维持在较高水平,而α-淀粉酶抑制率在消化后显著降低(P<0.05)。经消化后,核桃内种皮多酚的降血糖活性下降,但仍保留一定活性。
通过分析消化过程中具有潜在降血糖活性的酚类化合物的相对含量变化趋势,可探究核桃内种皮多酚的降血糖活性(表2)。高车前素[34]、异绿原酸C(4,5)[39]、槲皮素3-O-葡萄糖酸苷[40]、草质素[41]、香叶木素[42]等酚类化合物的相对含量变化趋势与α-葡萄糖苷酶抑制率一致,表明其参与了α-葡萄糖苷酶的调节,对核桃内种皮多酚的降血糖活性有重要贡献。酚类化合物中官能团的数量和位置是其抑制α-葡萄糖苷酶活性的关键因素,例如高车前素结构中的羟基能与底物竞争酶的催化位点,同时其疏水骨架能够诱导酶发生构象变化抑制α-葡萄糖苷酶的活性,且抑制类型为混合型抑制[43];异绿原酸C(4,5)具有抑制α-葡萄糖苷酶和蛋白酪氨酸磷酸酶1B(PTP1B)的作用,其结构中的两个咖啡酰基团可能是导致其抑制α-葡萄糖苷酶和PTP1B活性增强的原因[44];香叶木素结构上的羟基、甲氧基可能通过氢键与α-葡萄糖苷酶的活性位点结合,导致α-葡萄糖苷酶活性降低。因此这些酚类物质可能通过不同的潜在降血糖机制共同发挥抑制作用[42]。金合欢素、金圣草黄素、杨梅素等酚类化合物的相对含量变化呈下降趋势,与α-淀粉酶抑制率变化一致,表明其可能是核桃内种皮多酚中抑制α-淀粉酶的关键成分[45−47]。具有潜在降血糖活性的不同酚类物质可能通过结合α-淀粉酶的不同靶点抑制酶的活性,且可能随着这些物质的相对含量变化对α-淀粉酶的抑制能力也发生变化。例如,金圣草黄素通过调节碳水化合物代谢酶来恢复肝脏损伤和控制β细胞的降解,对链脲佐菌素(STZ)诱导的Ⅱ型糖尿病有显著作用[46];杨梅素能够通过两个氢键与α-淀粉酶的活性位点结合或与其他活性位点形成范德华力,从而抑制其酶活性[48]。综上,核桃内种皮多酚中的不同酚类化合物可能在消化过程中共同发挥活性,增强核桃内种皮多酚对两种酶的抑制效果。此前的研究也有类似报道,Zeng等[49]发现酚类物质在低浓度下发生相互作用对抑制α-葡萄糖苷酶表现出协同作用,Wang等[50]在探究结构相似的黄酮类物质抑制α-淀粉酶的机理研究及协同作用中提到芹菜素与黄芩素、高车前素、荆芥素联合使用,协同改善了它们对α-淀粉酶的抑制作用,这些物质间的相互作用进一步降低了酶的活性。
相关性分析是指对两个或多个具备相关性的变量元素进行分析,衡量两个变量因素的相关密切程度,相关系数的绝对值越大,表明因子之间的相关性越大[21]。图5为核桃内种皮多酚消化前后总酚含量与抗氧化能力、降血糖活性的相关性分析结果。在体外模拟消化中核桃内种皮多酚的总酚含量与DPPH∙清除率和ABTS+∙清除率均呈显著正相关(P<0.05,P<0.01),这与石志娇等[21]的研究结果类似,即多酚类物质可通过酚羟基作为氢供体直接与自由基反应、抑制产生自由基所需的酶活性、螯合诱导自由基产生的金属离子表现抗氧化活性[51]。此外,总酚含量与ABTS+∙清除率的相关性更高,存在极强的相关性(R2=0.996,P<0.01),原因为ABTS+∙两亲的结构可通过氢转移和转移电子实现自由基清除,而DPPH∙清除主要是通过抗氧化剂将氢转移给自由基[52]。各消化阶段核桃内种皮多酚的总酚含量与α-葡萄糖苷酶抑制率呈正相关(R2=0.939),表明多酚类物质可能是核桃内种皮发挥α-葡萄糖苷酶抑制活性的关键物质。而核桃内种皮多酚的总酚含量与α-淀粉酶抑制率的相关性较弱(R2=0.649),表明除酚类物质外还可能涉及其他物质发挥抑制作用。此外,α-葡萄糖苷酶抑制率与α-淀粉酶抑制率的皮尔逊相关系数仅为0.496,说明抑制这两种酶的多酚单体物质可能存在差异,这与前文中降血糖物质的分析结果一致。ABTS+∙清除率与α-葡萄糖苷酶抑制率呈显著正相关(R2=0.966,P<0.05),表明核桃内种皮多酚可能同时发挥抗氧化和降血糖活性并通过抗氧化应激和抑制糖消化酶共同发挥降血糖作用。以上结果表明总酚含量对核桃内种皮多酚的抗氧化能力和抑制α-葡萄糖苷酶有重要贡献,且其抗氧化与降血糖活性间存在一定的相关性,这为深入挖掘核桃内种皮多酚作为功能性食品原料的潜力提供了理论依据。
本研究以核桃内种皮为研究对象,通过超声辅助乙醇法提取其多酚,探究核桃内种皮多酚在体外模拟消化中总酚含量、酚类物质组成、抗氧化能力和降血糖活性的变化。研究发现,核桃内种皮多酚经消化后总酚含量显著降低(由695.27 mg GAE/g降至437.58 mg GAE/g),与口腔和胃消化阶段相比,其在小肠阶段降解较多;通过UHPLC-MS/MS技术共鉴定出224种酚类物质,主要为没食子酸、柯里拉京、绿原酸等;消化后核桃内种皮多酚的抗氧化能力和降血糖活性均下降,但其仍保留了一定的生物活性;消化过程中核桃内种皮多酚的总酚含量与抗氧化能力和降血糖活性的变化呈正相关,与α-淀粉酶抑制率相比,总酚含量与抗氧化能力和α-葡萄糖苷酶抑制率的线性相关性更强。因此,体外模拟消化过程影响核桃内种皮多酚的总酚含量及抗氧化能力和抑制糖消化酶活性,后续还需对核桃内种皮中的单体酚类物质进行实验,探究不同酚类物质之间的协同效应,明晰核桃内种皮多酚的抗氧化能力和降血糖活性,并利用动物模型探究核桃内种皮多酚在体内的转化机制及活性作用机理,为核桃内种皮的进一步开发利用提供实验依据。

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2026年第47卷第16期
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doi: 10.13386/j.issn1002-0306.2025070351
  • 接收时间:2025-07-30
  • 首发时间:2026-10-24
  • 出版时间:2026-08-15
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  • 收稿日期:2025-07-30
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    1西南林业大学生物与食品工程学院,云南昆明 650224
    2云南省高校大健康类森林资源开发利用工程研究中心,云南昆明 650224
    3云南省森林灾害预警与控制实验室,云南昆明 650224

通讯作者:

韩龙(1987−),男,博士,助理研究员,研究方向:食品微生物生态、发酵微生物、食品加工副产物与农林废弃物资源化利用,E-mail:。
范方宇(1979−),男,博士,教授,研究方向:农林产品加工,E-mail:。
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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