Article(id=1320176139079893550, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320175721343987874, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025060257, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1750348800000, receivedDateStr=2025-06-20, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792684119243, onlineDateStr=2026-10-22, pubDate=1781452800000, pubDateStr=2026-06-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792684119243, onlineIssueDateStr=2026-10-22, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792684119243, creator=13701087609, updateTime=1792684119243, updator=13701087609, issue=Issue{id=1320175721343987874, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='12', pageStart='1', pageEnd='517', issueExtLink='null', onlineDate='null', pubDate='1781452800000', pubDateStr='2026-06-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792684019647, creator='13701087609', updateTime=1792684019647, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=455, endPage=461, ext={EN=ArticleExt(id=1320176139247665711, articleId=1320176139079893550, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Lipid-lowering Activity and Mechanisms of Artemisia argyi Leaf Flavonoids Extract, columnId=1261343846965830077, journalTitle=Science and Technology of Food Industry, columnName=Nutrition and Healthcare, runingTitle=null, highlight=null, articleAbstract=

In this study, the flavonoids of Artemisia argyi leaf were extracted and identified. Subsequently, the lipid-lowering effects of Artemisia argyi leaf flavonoids extract (AALE) and its underlying mechanisms were investigated using a combination of the Caenorhabditis elegans (C. elegans) model and in vitro lipase inhibition assays. The results showed that AALE exhibited significant inhibitory activity against pancreatic lipase, with a half-maximal inhibitory concentration (IC50) of 8.085±0.28 mg/mL, and this inhibition was non-competitive. Furthermore, AALE significantly reduced triglyceride (TG) levels in a high-fat nematode model (P<0.05). The TG content in the high-fat control group was 2.35 mmol/g, whereas in the treatment groups supplemented with 240 and 360 µg/mL of AALE, the TG levels decreased 37.45% and 34.04% (P<0.05), respectively. Further studies revealed that AALE significantly downregulated the expression of key genes involved in fat synthesis pathways in nematodes, including fat-6, fat-7, pod-2, and fasn-1, while upregulating the expression of acs-2, a gene associated with lipolysis. Moreover, the effect of AALE in reducing TG levels was dependent on the critical fatty acid synthesis genes fat-6 and fat-7. In conclusion, AALE can inhibit pancreatic lipase activity and reduce the TG content in nematodes, demonstrating a good lipid-lowering effect. This study provides a theoretical basis for the high-value utilization of Artemisia argyi.

, authors=Rong LI1, *, Liang ZHANG1, Deyan ZHU1, Xianghui KONG2, Yechuan HUANG1, Wenjie DUAN3, authorsList=Rong LI, Liang ZHANG, Deyan ZHU, Xianghui KONG, Yechuan HUANG, Wenjie DUAN, authorCompany=null, correspAuthors=Rong LI, 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=1320176142192067134, articleId=1320176139079893550, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=蕲艾叶黄酮提取物降脂活性及作用机制, columnId=1261343850979779009, journalTitle=食品工业科技, columnName=营养与保健, runingTitle=null, highlight=null, articleAbstract=

本研究以蕲艾叶为研究对象,提取其中的黄酮类物质,并利用秀丽线虫模型和体外胰脂肪酶抑制活性探究其降脂效果及作用机制。结果表明,蕲艾叶黄酮提取物对胰脂肪酶具有显著的抑制活性,其半数抑制浓度IC50为8.085±0.28 mg/mL,且这种抑制作用为非竞争性抑制。此外,蕲艾叶黄酮提取物能显著降低高脂线虫模型体内甘油三酯(TG)含量(P<0.05),高脂对照组线虫体内TG含量为2.35 mmol/g,添加了240、360 µg/mL蕲艾叶黄酮提取物的高脂加药组线虫体内TG含量分别降低了37.45%、34.04%。进一步研究发现,蕲艾叶黄酮提取物显著下调了fat-6、fat-7、pod-2、fasn-1等线虫脂肪合成途径相关基因的表达,上调了脂肪分解有关基因acs-2的表达,且蕲艾叶黄酮提取物降低线虫体内TG含量依赖于脂肪酸合成关键基因fat-6、fat-7。综上,蕲艾叶黄酮提取物能够抑制胰脂肪酶活性,降低线虫体内TG含量,具有较好的降脂效果,本研究为蕲艾的高值化利用提供了理论依据。

, authors=李蓉1, *, 张亮1, 朱德艳1, 孔祥辉2, 黄业传1, 段文杰3, authorsList=李蓉, 张亮, 朱德艳, 孔祥辉, 黄业传, 段文杰, authorCompany=null, correspAuthors=李蓉, authorNote=

李蓉(1987−),女,博士,副教授,研究方向:天然产物研究与开发,E-mail:

, correspAuthorsNote=
李蓉(1987−),女,博士,副教授,研究方向:天然产物研究与开发,E-mail:。
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注:不同小写字母表示有显著性差异(P<0.05),图4同。

, figureFileSmall=NUBD8/eo3IYknPpl1S/eWQ==, figureFileBig=/34cfAowWQpIaEAPFMgLMQ==, tableContent=null), ArticleFig(id=1320176147078431350, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=EN, label=Fig.4, caption=Effect of AALE on the relative quanti4cation of fat contents by Oil-Red-O staining in wild-type worms, figureFileSmall=W8Wi6HRKfhfw4AVoGYj38g==, figureFileBig=LRCgf53redlpoLEmFlQRbQ==, tableContent=null), ArticleFig(id=1320176147149734519, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=CN, label=图4, caption=蕲艾叶黄酮提取物对野生型线虫油红O染色定量结果的影响

注:A. 不同处理组野生型线虫油红O染色代表性图片;B. 不同处理组野生型线虫油红O染色定量结果。

, figureFileSmall=W8Wi6HRKfhfw4AVoGYj38g==, figureFileBig=LRCgf53redlpoLEmFlQRbQ==, tableContent=null), ArticleFig(id=1320176147242009208, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=EN, label=Fig.5, caption=Effect of AALE on the expression of lipid metabolism-related genes in wild-type nematodes, figureFileSmall=EPkRD5N41ls7Z30VQ03ISw==, figureFileBig=0KQA+8eo7itkkykeFK++3g==, tableContent=null), ArticleFig(id=1320176147317506681, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=CN, label=图5, caption=蕲艾叶黄酮提取物对野生型线虫脂代谢相关基因表达的影响

注:结果用平均值±标准偏差表示,*P<0.05,**P<0.01表示与高脂对照组比较。

, figureFileSmall=EPkRD5N41ls7Z30VQ03ISw==, figureFileBig=0KQA+8eo7itkkykeFK++3g==, tableContent=null), ArticleFig(id=1320176147439141498, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=EN, label=Fig.6, caption=Effect of AALE on the TG content of BX106 strains, figureFileSmall=mHrAQORm9A7mrj0pI/tleA==, figureFileBig=w9pziUJHV4IbkyBjAFH/fg==, tableContent=null), ArticleFig(id=1320176147543999099, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=CN, label=图6, caption=蕲艾叶黄酮提取物对BX106线虫株系TG含量的影响

注:ns表示与高脂对照组无显著差异,图7同。

, figureFileSmall=mHrAQORm9A7mrj0pI/tleA==, figureFileBig=w9pziUJHV4IbkyBjAFH/fg==, tableContent=null), ArticleFig(id=1320176147623690876, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=EN, label=Fig.7, caption=Effect of AALE on the TG content of BX153 strains, figureFileSmall=oQVi0nH+Fv3So1jubwy4Gw==, figureFileBig=hVQNRGrGao0d9NG8E/3z/w==, tableContent=null), ArticleFig(id=1320176147682411133, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=CN, label=图7, caption=蕲艾叶黄酮提取物对BX153线虫株系TG含量的影响, figureFileSmall=oQVi0nH+Fv3So1jubwy4Gw==, figureFileBig=hVQNRGrGao0d9NG8E/3z/w==, tableContent=null), ArticleFig(id=1320176147753714302, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=EN, label=Table 1, caption=

Reaction system for determination of pancreatic lipase inhibitory activity

, figureFileSmall=null, figureFileBig=null, tableContent=
组别PBS缓冲液
(μL)
胰脂肪酶溶液
(μL)
样品溶液
(μL)
底物溶液
(μL)
对照试验组10050050
对照空白组1500050
样品试验组50505050
样品空白组10005050
), ArticleFig(id=1320176147841794687, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=CN, label=表1, caption=

胰脂肪酶抑制活性反应体系

, figureFileSmall=null, figureFileBig=null, tableContent=
组别PBS缓冲液
(μL)
胰脂肪酶溶液
(μL)
样品溶液
(μL)
底物溶液
(μL)
对照试验组10050050
对照空白组1500050
样品试验组50505050
样品空白组10005050
), ArticleFig(id=1320176147917292160, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=EN, label=Table 2, caption=

Primer sequences of related genes

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基因引物
act-1F:CATGAAGATCAAGATCATCGCC
R:GTGACGATGGTTTTGAACTTGT
fat-6F:TCAACAGCGCTGCTCACTAT
R:TTCGACTGGGGTAATTGAGG
fat-7F:CAACAGCGCTGCTCACTATT
R:CACCAACGGCTACAACTGTG
acs-2F:GCCTTGGATGGGATAGAG
R:TGATGGGAAGACCACAGT
pod-2F:GTAAATCGAGTCAATACCGTGC
R:GCGAGATTTTCTTCTGAACCTC
fasn-1F:TCATGGTTGATAGAGACTACGC
R:AACCAAAGCATAATAAGCGGTG
), ArticleFig(id=1320176147984401025, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320176139079893550, language=CN, label=表2, caption=

相关基因引物序列

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基因引物
act-1F:CATGAAGATCAAGATCATCGCC
R:GTGACGATGGTTTTGAACTTGT
fat-6F:TCAACAGCGCTGCTCACTAT
R:TTCGACTGGGGTAATTGAGG
fat-7F:CAACAGCGCTGCTCACTATT
R:CACCAACGGCTACAACTGTG
acs-2F:GCCTTGGATGGGATAGAG
R:TGATGGGAAGACCACAGT
pod-2F:GTAAATCGAGTCAATACCGTGC
R:GCGAGATTTTCTTCTGAACCTC
fasn-1F:TCATGGTTGATAGAGACTACGC
R:AACCAAAGCATAATAAGCGGTG
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蕲艾叶黄酮提取物降脂活性及作用机制
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李蓉 1, * , 张亮 1 , 朱德艳 1 , 孔祥辉 2 , 黄业传 1 , 段文杰 3
食品工业科技 | 营养与保健 2026,47(12): 455-461
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食品工业科技 |营养与保健 2026 , 47 (12) : 455 -461
蕲艾叶黄酮提取物降脂活性及作用机制
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李蓉1, * , 张亮1, 朱德艳1, 孔祥辉2, 黄业传1, 段文杰3
作者信息
  • 1荆楚理工学院特色食品功能挖掘与综合利用团队,湖北荆门 448000
  • 2荆门市中心医院,湖北荆门 448001
  • 3湖北仙之灵食品有限公司,湖北荆门 431935
通讯作者:
李蓉(1987−),女,博士,副教授,研究方向:天然产物研究与开发,E-mail:。
作者简介:

李蓉(1987−),女,博士,副教授,研究方向:天然产物研究与开发,E-mail:

Lipid-lowering Activity and Mechanisms of Artemisia argyi Leaf Flavonoids Extract
Rong LI1, * , Liang ZHANG1, Deyan ZHU1, Xianghui KONG2, Yechuan HUANG1, Wenjie DUAN3
Affiliations
  • 1Functional Exploration and Comprehensive Utilization Team for Specialty Foods, Jingchu University of Technology, Jingmen 448000, China
  • 2Jingmen Central Hospital of Jingchu University of Technology, Jingmen 448001, China
  • 3Hubei Xianzhiling Foods Corporation, Jingmen 431935, China
出版时间: 2026-06-15 doi: 10.13386/j.issn1002-0306.2025060257
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本研究以蕲艾叶为研究对象,提取其中的黄酮类物质,并利用秀丽线虫模型和体外胰脂肪酶抑制活性探究其降脂效果及作用机制。结果表明,蕲艾叶黄酮提取物对胰脂肪酶具有显著的抑制活性,其半数抑制浓度IC50为8.085±0.28 mg/mL,且这种抑制作用为非竞争性抑制。此外,蕲艾叶黄酮提取物能显著降低高脂线虫模型体内甘油三酯(TG)含量(P<0.05),高脂对照组线虫体内TG含量为2.35 mmol/g,添加了240、360 µg/mL蕲艾叶黄酮提取物的高脂加药组线虫体内TG含量分别降低了37.45%、34.04%。进一步研究发现,蕲艾叶黄酮提取物显著下调了fat-6、fat-7、pod-2、fasn-1等线虫脂肪合成途径相关基因的表达,上调了脂肪分解有关基因acs-2的表达,且蕲艾叶黄酮提取物降低线虫体内TG含量依赖于脂肪酸合成关键基因fat-6、fat-7。综上,蕲艾叶黄酮提取物能够抑制胰脂肪酶活性,降低线虫体内TG含量,具有较好的降脂效果,本研究为蕲艾的高值化利用提供了理论依据。

蕲艾叶  /  黄酮类  /  胰脂肪酶  /  秀丽隐杆线虫  /  降脂

In this study, the flavonoids of Artemisia argyi leaf were extracted and identified. Subsequently, the lipid-lowering effects of Artemisia argyi leaf flavonoids extract (AALE) and its underlying mechanisms were investigated using a combination of the Caenorhabditis elegans (C. elegans) model and in vitro lipase inhibition assays. The results showed that AALE exhibited significant inhibitory activity against pancreatic lipase, with a half-maximal inhibitory concentration (IC50) of 8.085±0.28 mg/mL, and this inhibition was non-competitive. Furthermore, AALE significantly reduced triglyceride (TG) levels in a high-fat nematode model (P<0.05). The TG content in the high-fat control group was 2.35 mmol/g, whereas in the treatment groups supplemented with 240 and 360 µg/mL of AALE, the TG levels decreased 37.45% and 34.04% (P<0.05), respectively. Further studies revealed that AALE significantly downregulated the expression of key genes involved in fat synthesis pathways in nematodes, including fat-6, fat-7, pod-2, and fasn-1, while upregulating the expression of acs-2, a gene associated with lipolysis. Moreover, the effect of AALE in reducing TG levels was dependent on the critical fatty acid synthesis genes fat-6 and fat-7. In conclusion, AALE can inhibit pancreatic lipase activity and reduce the TG content in nematodes, demonstrating a good lipid-lowering effect. This study provides a theoretical basis for the high-value utilization of Artemisia argyi.

Artemisia argyi leaf  /  flavonoid  /  pancreatic lipase  /  Caenorhabditis elegans  /  lipid-lowering
李蓉, 张亮, 朱德艳, 孔祥辉, 黄业传, 段文杰. 蕲艾叶黄酮提取物降脂活性及作用机制. 食品工业科技, 2026 , 47 (12) : 455 -461 . DOI: 10.13386/j.issn1002-0306.2025060257
Rong LI, Liang ZHANG, Deyan ZHU, Xianghui KONG, Yechuan HUANG, Wenjie DUAN. Lipid-lowering Activity and Mechanisms of Artemisia argyi Leaf Flavonoids Extract[J]. Science and Technology of Food Industry, 2026 , 47 (12) : 455 -461 . DOI: 10.13386/j.issn1002-0306.2025060257
肥胖及其并发症已成为一个重大的公共卫生问题。肥胖是由能量摄入与能量消耗之间的失衡所导致的,脂代谢功能障碍极大地影响了能量平衡,从而导致脂肪过度堆积。因此,具有调节脂质代谢的天然产物引起人们广泛的关注。研究表明天然产物如红小豆提取物[1]、枸杞多糖[2]、韭菜提取物[3]等对胰脂肪酶具有较好的抑制活性,显示具有降脂功效的潜力。蕲艾(Artemisia argyi Levl. et Vant.)是产自湖北蕲春的一种菊科蒿属艾的栽培品种,具有重要的药用价值和经济价值。蕲艾叶中含有黄酮类、鞣酸类、挥发油、有机酸类等多种化合物,其中黄酮类化合物是其具有生物活性的重要物质基础[4]。目前关于蕲艾黄酮化合物生物活性研究主要集中在抗氧化、抗炎、增强免疫等方面[5−7],对其调节脂代谢及其作用机制方面的报道较少。最近的研究显示在饲料中添加一定量的艾草粉可以调节湖羊的脂代谢[8],表明蕲艾具有潜在的降脂活性。
秀丽隐杆线虫(Caenorhabditis elegans)因其体型小、生命周期短、身体透明、遗传操作简便以及培养成本低廉等特点,已成为研究脂质代谢和肥胖遗传学的热门模型[9−10]。更重要的是,许多调节人类脂代谢的关键信号通路和基因在秀丽隐杆线虫中是保守的[11]。胰脂肪酶是人体消化系统中脂肪分解的关键酶,抑制胰脂肪酶活性可以抑制机体对脂肪的吸收[12]。基于此,本研究以蕲艾叶黄酮提取物为研究对象,测定其对胰脂肪酶的抑制活性,再利用秀丽线虫模型探究蕲艾叶黄酮提取物体内降脂效果,并对其降脂作用的分子机制进行探究,为蕲艾叶在降脂性功能食品开发方面提供理论依据。
蕲艾叶 购于蕲春蕲艾康本草科技有限公司;野生型秀丽线虫株系(N2)、BX153 [fat-7(wa36) Ⅴ]、BX106 [fat-6(tm331) Ⅳ]、E. coli OP50 购于明尼苏达大学秀丽隐杆线虫遗传学中心Caenorhabditis Genetics Center(CGC);胰脂肪酶(15~35 U/mg)、4-硝基苯基丁酸酯、奥利司他、芦丁标准品(纯度≥98%)、油红O粉末 上海阿拉丁生化科技股份有限公司;葡萄糖、亚硝酸钠、氯仿、无水乙醇、异丙醇、无水氯化钙、七水合硫酸亚铁、四水合氯化锰、七水合硫酸锌、五水合硫酸铜、十二水合磷酸氢二钠、胆固醇、次氯酸钠、七水合硫酸镁、氢氧化钠、氯化钠、磷酸二氢钾、磷酸氢二钾 国药集团;5-氟-2-脱氧尿苷(FudR) 美国Sigma公司;Trizol 试剂、SuperReal PreMix Plus(SYBR Green) 北京天根生化科技有限公司;cDNA反转录试剂盒、qPCR试剂盒SYBR 美国诺唯赞公司;TG检测试剂盒、BCA检测试剂盒 南京建成生物工程研究所。
SW-CJ-3F超净工作台 上海沪静医疗器械有限公司;NIB610倒置显微镜 宁波永新光学股份有限公司;XFH-50CA全自动高压灭菌锅 浙江新丰医疗器械有限公司;Multiskan Skyhigh全波长酶标仪 美国Thermo公司;MY-10手持研磨器 上海净信实业发展有限公司;Centrifuge 5418 R冷冻离心机 德国艾本德Eppendorf公司;QuantStudio 3.0荧光定量PCR仪 美国ABI公司。
参照Shi等[13]方法提取蕲艾叶中的黄酮化合物。蕲艾叶在60 ℃下烘干后粉碎,按照料液比1:15以50%乙醇溶液在50 ℃下提取2 h,重复浸提2次后抽滤,所得滤液在50 ℃下旋转蒸发去除乙醇得到浓缩液,将浓缩液经AB-8大孔树脂纯化,收集50%乙醇洗脱液,浓缩并冷冻干燥得到蕲艾叶黄酮提取物粉末。用DMSO溶解粗提物粉末配成浓度为60 mg/mL的母液,在−80 ℃保存备用。
采用亚硝酸钠-硝酸铝法测定总黄酮含量[14]。将蕲艾叶黄酮提取物冻干粉溶解于50%乙醇中,取200 μL不同浓度的芦丁标准品溶液或样品溶液,加入0.5% NaNO2溶液75 μL混匀后,避光静置5 min,再加入150 μL 10% Al(NO3)3·6H2O溶液,静置5 min,最后加入1 mol/L NaOH溶液500 μL,在λ=510 nm处测定吸光值,根据芦丁标准曲线(y=0.0019x+0.0309,R2=0.9931)和样品溶液的吸光度计算样品中的总黄酮含量,结果以mg芦丁/g样品表示。
参考缪园欣等[15]方法并稍作改进。按照表1反应体系进行实验,取50 μL不同浓度样品溶液(5、10、15、20、25 mg/mL)、50 μL胰脂肪酶(10 mg/mL)、以及不同体积的PBS磷酸盐缓冲液(50 mmol/L,pH8.0)混匀后于37 ℃恒温孵育15 min,加入50 μL 1 mg/mL 4-硝基苯基丁酸酯溶液,混匀后在37 ℃继续孵育15 min。使用酶标仪在405 nm下测定吸光度,以奥利司他作为阳性对照,按照式(1)计算脂肪酶抑制率。
$\rm 抑制率(\text{%})= \left(1-\frac{\mathrm{A}1-\mathrm{A}0}{\mathrm{B}1-\mathrm{B}0}\right)\times 100 $
式中:A1为样品试验组吸光度值;A0为样品空白组吸光度值;B1为对照试验组吸光度值;B0为对照空白组吸光度值。
固定胰脂肪酶浓度为10 mg/mL,分别与质量浓度为0、20 mg/mL的样品反应,按照1.2.3反应体系分别测定不同浓度的样品在不同底物4-硝基苯基丁酸酯浓度(2、4、6、8、10 mg/mL)下的反应初速度,以反应速率的倒数(1/[V])对底物4-硝基苯基丁酸酯质量浓度的倒数(1/[S])作图,绘制Lineweaver-Burk双倒数曲线,确定样品对胰脂肪酶的抑制类型。
将处于产卵期的线虫用M9缓冲液从线虫培养板上洗入1.5 mL离心管中,去除上清液,再用M9缓冲液洗三次除去线虫身上粘附的菌。然后在离心管中加入1 mL线虫裂解液(由氢氧化钠溶液与次氯酸钠混合配制),迅速剧烈振荡10 min至虫体完全裂解,再在10000 r/min常温离心1 min,弃去上清,并用M9缓冲液清洗沉淀4次,最后将得到的虫卵置于涂有E. coli OP50的线虫培养皿上,放在20 ℃培养箱中培养,获得同步化的线虫子代[16]。
参照Peng等[17]的方法建立高脂线虫模型:用24孔板培养,每孔加入300 µL线虫液体培养基(实验室自制),其中设置仅含有DMSO的空白对照组、含有10 mmol/L葡萄糖的高糖对照组以及含有10 mmol/L葡萄糖和不同浓度(60、240、360 µg/mL)蕲艾叶黄酮提取物的高糖加药组。将同步化至L4期的线虫用挑针挑取至线虫液体培养基中,每孔挑取约300条线虫,挑取线虫的时间设为第0 d,密封24孔板后放入20 ℃培养。
参照1.2.5.2方法培养不同株系线虫包括野生型N2、BX106、BX153,当培养3 d后,收集不同处理组线虫,用M9缓冲液洗3次去除粘附的杂菌,在冰上充分研磨,再在4 ℃下10000 r/min离心5 min,按照甘油三酯TG、BCA测定试剂盒说明书测定线虫匀浆液上清中TG、蛋白质含量,每个实验重复三次。
利用油红O染色法观察线虫体内脂肪含量及分布情况。方法如下:线虫的培养参照1.2.5.2,当培养至第3 d时,收集不同处理组的N2线虫于1.5 mL离心管中,用M9缓冲液洗三次,加入2%多聚甲醛100 μL固定,然后将离心管放入液氮中,反复冻融三次,最后一次解冻后于4 ℃下1500 r/min离心1 min,用M9缓冲液洗2次,去除上清后加入1 mL 60%异丙醇脱水15 min,离心去上清,再加入1 mL 60%油红O染色液,振荡过夜,避光染色。染色后取出离心管,4 ℃下1500 r/min离心1 min,除去染色液,再加M9洗2次除去多余的浮色,将线虫转移至载玻片上,在显微镜下观察线虫脂滴染色情况,并用ImageJ软件处理数据[18]。
按照1.2.5.2所述方法培养野生型线虫N2,其中设置仅含有DMSO的高糖对照组和含有240 μg/mL蕲艾叶黄酮提取物的高糖加药组。待线虫培养至成虫第3 d,收集不同处理组的线虫用M9缓冲液洗3次去除粘附的杂菌,在冰上充分研磨,再在4 ℃下10000 r/min离心5 min得到上清液。参照试剂盒说明书,利用Trizol提取法获得高质量的总RNA,再按照逆转录试剂盒说明书,配制反转录反应体系,混匀后37 ℃孵育15 min,85 ℃孵育5 s,获得的cDNA。根据SYBR Premix EX TapⅡ说明书进行定量扩增,反应体系按照5 μL 2×qPCR Mix,0.4 μL正向引物,0.4 μL反向引物,0.5 μL cDNA,3.7 μL ddH2O配制反应体系,在荧光定量PCR仪中进行反应。设定反应程序为:95 ℃预变性30 s,95 ℃变性5 s,57 ℃退火30 s,72 ℃延伸30 s,40个循环。以act-1作为内参基因,采用2−∆∆CT法计算目的基因的相对表达量。相关基因引物由武汉擎科生物有限公司合成,具体引物序列见表2。
所有实验重复三次,结果以均值±标准偏差表示。实验数据用SPSS进行统计分析,采用ANOVA进行显著性分析,P<0.05表示具有显著性差异。
对蕲艾叶黄酮提取物冻干粉中的黄酮含量进行测定,其黄酮含量为176.25±1.38 mg芦丁/g。在前期的研究成果中[19],该蕲艾叶黄酮提取物中共鉴定出22种黄酮类化合物,其中以山柰酚、芦丁、芹菜素、儿茶素、槲皮素、木犀草素、刺芒柄花素和柚皮素等8种黄酮类化合物为主。
由图1可知,蕲艾叶黄酮提取物AALE浓度为5~25 mg/mL范围内,随着浓度的增加,其对胰脂肪酶的抑制率显著增加,且具有一定的量效关系,当其浓度为25 mg/mL时,其对胰脂肪酶的抑制率为77.19%,略低于阳性对照奥利司他的抑制率91.28%,这可能与黄酮类化合物可以与胰脂肪酶的活性部位通过疏水缔合或分子间氢键形成相互作用,从而改变酶的结构并抑制其活性[20]。经过计算AALE对胰脂肪酶的半数抑制浓度IC50为8.085±0.28 mg/mL,奥利司他对胰脂肪酶的半数抑制浓度IC50为1.068±0.13 mg/mL,结果表明在一定的质量浓度下,AALE对胰脂肪酶抑制效果较好,显示其降脂的潜力。
固定胰脂肪酶浓度和样品质量浓度,改变反应体系中的底物浓度,以反应初速度的倒数1/[V]对底物浓度的倒数1/[S]绘制Lineweaver-Burk双倒数图,当对照组曲线与样品组曲线相交于横轴,则为非竞争性抑制[1−2]。如图2所示,对照组与AALE试验组拟合曲线相交于横轴,表明AALE对胰脂肪酶的抑制作用为非竞争性抑制。
本研究通过在秀丽线虫培养基中添加10 mmol/L葡萄糖建立高脂线虫模型,培养3 d后测定秀丽线虫体内甘油三酯TG含量,结果如图3所示。与对照组相比,添加了10 mmol/L葡萄糖的线虫高糖组体内TG含量显著增加,由对照组的0.96 mmol/g增加至2.35 mmol/g,这与Zhang等[21]报道结果一致,其添加了10 mmol/L葡萄糖后,高脂对照组线虫体内TG含量为空白对照组的1.5倍,表明本研究成功建立了高脂线虫模型。与高脂对照组HF相比,添加了60、240、360 µg/mL蕲艾叶黄酮提取物的高脂加药组线虫体内TG含量分别降低了16.17%、37.45%、34.04%,其中240 µg/mL蕲艾叶黄酮提取物降脂效果最佳,因此后续实验选择240 µg/mL这一浓度。与本研究结果类似,研究报道黄酮类化合物芦丁、木犀草素、槲皮素、芹菜素等能显著降低秀丽线虫体内的脂肪含量[22],富含黄酮类化合物的银杏种子提取物也对秀丽线虫脂代谢由积极的调节作用[23]。
糖代谢与脂代谢紧密相关,过量的糖摄入会导致脂代谢紊乱,诱导肥胖及相关疾病的发生。与哺乳动物类似,秀丽线虫摄入过量的糖后会在体内以脂滴的形式贮存,其主要分布在肠道及皮下细胞中[24]。油红O是一种脂溶性染料,可以进入细胞将脂滴染成红色,由于线虫身体透明,因此可以直观地观察染料的颜色及位置并以此判断线虫体内脂肪含量及分布情况[25−26]。为了观察蕲艾叶黄酮提取物对秀丽线虫体内脂肪含量的影响,本研究对野生型秀丽线虫体内的脂肪进行了油红O染色,结果如图4所示。与空白对照组相比,高脂对照组线虫虫体红色明显加深,数据处理显示脂肪含量显著提高了28.86%。当加入240 µg/mL蕲艾叶黄酮提取物共同培养后,虫体红色比高脂对照组要浅,数据处理显示与高脂对照组相比,240 µg/mL蕲艾叶黄酮提取物处理组其脂肪含量降低了14.73%,此结果更加直观地显示蕲艾叶黄酮提取物可以显著降低线虫体内脂肪含量,这一结果与上述TG结果一致,表明蕲艾叶提取物具有较好的体内降脂效果。
为了探究蕲艾叶黄酮提取物对线虫降脂作用的分子机制,本研究测定了蕲艾叶黄酮提取物对线虫体内脂代谢相关基因表达的影响。如图5所示,与高脂对照组相比,240 µg/mL AALE处理后线虫fat-6、fat-7的表达显著下调(P<0.05),分别为对照组的0.64、0.38倍。在秀丽线虫中,fat-6和fat-7编码硬脂酰辅酶A去饱和酶,在硬脂酸转化为油酸的过程中发挥重要作用[27−28]。AALE显著下调fat-6、fat-7的表达,表明其抑制了不饱和脂肪酸的合成途径,从而对降低脂肪含量起到积极的作用。此外,与高脂对照组相比,240 µg/mL AALE显著下调了pod-2、fasn-1的表达(P<0.01),其分别为对照组的0.58、0.63倍。脂代谢基因pod-2编码线虫体内乙酰辅酶A羧化酶,是脂肪酸生物合成中的限速酶,它催化乙酰辅酶A作为底物生成丙二酰辅酶A,与线虫体内中长链脂肪酸生物合成有关[29]。而fasn-1编码脂肪酸合成酶,是脂肪酸生物合成中的另一种限速酶[30]。AALE显著下调pod-2、fasn-1的表达(P<0.01),表明其抑制了脂肪酸的合成途径,从而降低线虫体内脂肪含量。相反地,与高脂对照组相比,240 µg/mL AALE显著增加了acs-2的表达量(P<0.05),其表达水平为对照组的1.87倍。在秀丽线虫中,acs-2编码线粒体酰基辅酶A合成酶,催化脂肪酸转化为酰基辅酶A,可以促进脂肪酸转运至线粒体基质用于后续的β氧化[31−32]。AALE上调acs-2的表达,表明AALE有助于激活脂肪酸β氧化途径,通过促进脂肪分解途径而降低线虫体内脂肪的积累水平。综上所述,AALE通过抑制脂肪合成相关基因的表达,促进脂肪分解途径相关基因的表达,降低了线虫体内脂肪含量。
BX106线虫株系是fat-6缺失突变体,鉴于前面已经表明AALE显著降低线虫体内fat-6的表达水平,为了进一步探究fat-6在AALE调控的降脂效果中的作用,本研究测定了BX106线虫体内TG含量。如图6所示,AALE不能显著降低fat-6缺失突变体BX106株系的TG含量,表明AALE的降脂效果依赖于fat-6,这与AALE调节fat-6基因表达结果一致,fat-6在AALE的降脂效果中发挥重要作用。Zhang等[21]研究指出,五没食子酰葡萄糖可以显著降低野生型秀丽线虫体内TG含量,但是不能降低fat-6缺失突变体BX106线虫体内TG含量,表明五没食子酰葡萄糖降低秀丽线虫TG含量依赖于fat-6。
BX153线虫株系是fat-7缺失突变体,常用于活性成分降脂作用机制研究。Lin等[33]报道指出,苦瓜皂苷提取物能显著降低野生型秀丽线虫体内脂肪含量,但是不能降低fat-7缺失突变体线虫BX153体内脂肪含量,表明苦瓜皂苷提取物降低线虫脂肪含量依赖于fat-7。鉴于前面结果显示AALE显著降低线虫体内fat-7的表达水平,为了进一步探究fat-7在AALE调控的降脂效果中的作用,本研究测定了BX153线虫体内TG含量。如图7所示,AALE不能显著降低fat-7缺失突变体BX153株系的TG含量,而AALE能显著降低野生型线虫株系体内TG含量,表明AALE的降脂效果依赖于fat-7,其在AALE的降脂效果中发挥重要作用。
本研究提取了蕲艾叶中的黄酮类化合物并对其进行了鉴定,其含有山柰酚、芦丁、芹菜素、儿茶素、槲皮素、木犀草素、刺芒柄花素、柚皮素等为主的黄酮类物质。利用体外模型探究其降脂活性,结果表明其对胰脂肪酶具有良好的抑制作用,且为非竞争性抑制。此外,蕲艾叶黄酮提取物可以显著降低秀丽线虫体内甘油三酯含量,进一步研究发现其通过下调线虫体内脂肪合成途径相关基因的表达,上调脂肪分解途径相关基因的表达水平从而起到降脂效果。本研究通过探究蕲艾叶黄酮提取物体内外降脂活性,初步阐明其体内降脂作用的分子机制,为蕲艾叶在降脂方面的应用提供理论依据。

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2026年第47卷第12期
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doi: 10.13386/j.issn1002-0306.2025060257
  • 接收时间:2025-06-20
  • 首发时间:2026-10-22
  • 出版时间:2026-06-15
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    1荆楚理工学院特色食品功能挖掘与综合利用团队,湖北荆门 448000
    2荆门市中心医院,湖北荆门 448001
    3湖北仙之灵食品有限公司,湖北荆门 431935

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李蓉(1987−),女,博士,副教授,研究方向:天然产物研究与开发,E-mail:。
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2种不同金属材料的力学参数
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genus
种数
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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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