Article(id=1320888947337707780, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320883923090002610, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025050027, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1746633600000, receivedDateStr=2025-05-08, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792854065975, onlineDateStr=2026-10-24, pubDate=1774972800000, pubDateStr=2026-04-01, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792854065975, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792854065975, creator=13701087609, updateTime=1792854065975, updator=13701087609, issue=Issue{id=1320883923090002610, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='7', pageStart='1', pageEnd='510', issueExtLink='null', onlineDate='null', pubDate='1774972800000', pubDateStr='2026-04-01', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792852868101, creator='13701087609', updateTime=1792852868101, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=216, endPage=227, ext={EN=ArticleExt(id=1320888948939931909, articleId=1320888947337707780, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Screening of Cholesterol-lowering Lactic Acid Bacteria from Traditional Yak Yogurt on the Western Sichuan Plateau and Its Intervention in Hypercholesterolemic Mice, columnId=1261336275769016441, journalTitle=Science and Technology of Food Industry, columnName=Bioengineering, runingTitle=null, highlight=null, articleAbstract=

This study aimed to screen a strain with high cholesterol-lowering effect from traditional fermented yak yogurt in western Sichuan province and explore its therapeutic effect on hypercholesterolemia. This study focused on 295 strains of lactic acid bacteria isolated from traditional fermented yak yogurt in Western Sichuan, and the lactic acid bacteria with high cholesterol-lowering ability were screened by ferric ammonium sulfate method and o-phthalaldehyde method, and the in vitro probiotic characterization test and safety evaluation were carried out, and the cholesterol-lowering effect of the strains was evaluated by establishing a mouse model of hypercholesterolemia induced by high-fat diet. The screening revealed four strains exhibiting favorable cholesterol-reducing efficiency and efficacy isolated from 295 lactic acid bacteria. Among the 4 strains, strain 166 had the highest survival rate in simulated gastric fluid tolerance assay test, and the number of viable bacteria remained above 5.3×108 CFU/mL after 3 h of incubation in the simulated intestinal fluid, and it possessed a good bile salt tolerance performance, with a hydrophobic capacity of up to 62.3%, auto-aggregation capacity of 18.65%, which was identified as Limosilactobacillus fermentum. In addition, strain 166 was evaluated negatively for harmful metabolites, no hemolysis phenomenon, no gelatin liquefaction, and showed some resistance to eight antibiotics, which showed good in vitro probiotic properties and safety. In vivo experiments in animals showed that Limosilactobacillus fermentum 166 could significantly reduce the serum total cholesterol, triglycerides, low-density lipoprotein cholesterol levels and atherosclerosis index of mice, effectively increase the content of serum high-density lipoprotein cholesterol, and effectively improve the Lee's index, liver index, perirenal fat index and abdominal fat index. Therefore, Limosilactobacillus fermentum 166 has the potential to be developed as a functional probiotic product, providing data support for the prevention and adjunctive therapy of hypercholesterolemia.

, authors=Sirui WANG1, Xiumei REN2, Shiqi ZHANG1, Jing LI1, Yingjie WANG1, Yihao WAN1, Lianhong CHEN1, *, authorsList=Sirui WANG, Xiumei REN, Shiqi ZHANG, Jing LI, Yingjie WANG, Yihao WAN, Lianhong CHEN, authorCompany=null, correspAuthors=Lianhong CHEN, 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=1320888957760553255, articleId=1320888947337707780, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=川西高原传统牦牛酸乳中降胆固醇乳酸菌的筛选及其对高胆固醇血症小鼠的干预作用, columnId=1261336277547401341, journalTitle=食品工业科技, columnName=生物工程, runingTitle=null, highlight=null, articleAbstract=

本研究旨在从川西传统发酵牦牛酸乳中筛选出一株具有高效降胆固醇作用的菌株,探究其对高胆固醇血症的干预效果。研究聚焦于川西传统发酵牦牛酸乳中分离出的295株乳酸菌,采用硫酸铁铵法和邻苯二甲醛法筛选出降胆固醇能力较高的乳酸菌,并对其进行体外益生性试验和安全性评价,通过建立高脂饮食诱导的高胆固醇血症小鼠模型评估菌株降胆固醇效果。结果表明,从295株乳酸菌中筛选得到4株胆固醇降低效率和胆固醇降低效力良好的菌株,其中菌株166在人工胃液耐受性测定试验中为4株菌株中存活率最高的,在人工肠液中培养3 h后活菌数保持在5.3×108 CFU/mL以上,具有良好的胆盐耐受性能,疏水能力和自聚集力分别达到62.3%和18.65%,经鉴定其为发酵粘液乳杆菌。此外,菌株166的有害代谢产物评价结果均为阴性,无溶血现象,不溶解明胶,对8种抗生素均表现出一定抗性,表现出良好的体外益生性和安全性。动物体内实验表明,发酵粘液乳杆菌166能显著降低小鼠血清总胆固醇(Total cholesterol,TC)、甘油三酯(Triglyceride,TG)、低密度脂蛋白胆固醇(Low-density lipoprotein cholesterol,LDLC)水平及动脉粥样硬化指数(Atherosclerosis index,AI),有效提高血清高密度脂蛋白胆固醇(High-density lipoprotein cholesterol,HDLC)含量,并有效改善小鼠Lee’s指数、肝脏指数、肾周脂肪指数和腹部脂肪指数。因此,发酵粘液乳杆菌166具有开发为功能性益生菌产品的潜力,为干预高胆固醇血症提供数据支撑。

, authors=王思瑞1, 任秀梅2, 张诗琦1, 李静1, 王樱洁1, 宛祎豪1, 陈炼红1, *, authorsList=王思瑞, 任秀梅, 张诗琦, 李静, 王樱洁, 宛祎豪, 陈炼红, authorCompany=null, correspAuthors=陈炼红, authorNote=

王思瑞(2004−),女,本科,研究方向:食品加工,E-mail:

, correspAuthorsNote=
陈炼红(1967−),女,硕士,教授,研究方向:民族食品资源开发,E-mail:。
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王思瑞(2004−),女,本科,研究方向:食品加工,E-mail:

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注:不同小写字母表示组间差异显著(P<0.05),含相同字母表示差异不显著(P>0.05);图4~图5,图12~图16同。

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注:A:溶血性金黄色葡萄球菌;B:发酵粘液乳杆菌166号。

, figureFileSmall=rGwzAUXIjY3O0ejoE6fg9w==, figureFileBig=rkIGrPIj/owFCdvp9w4qXw==, tableContent=null), ArticleFig(id=1320888963892625775, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.10, caption=Strain 166 amino acid decarboxylase activity, figureFileSmall=8YHijiqK2hOJO2JYRgIK1w==, figureFileBig=agJDS8k/xcMobChZLFiyAQ==, tableContent=null), ArticleFig(id=1320888963989094768, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图10, caption=菌株166氨基酸脱羧酶活性, figureFileSmall=8YHijiqK2hOJO2JYRgIK1w==, figureFileBig=agJDS8k/xcMobChZLFiyAQ==, tableContent=null), ArticleFig(id=1320888964056203633, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.11, caption=Gelatin liquefaction results of strain 166, figureFileSmall=B0/yh08tVXayLyzCS3hivQ==, figureFileBig=/Je9WFev0g4oElrPDevtpA==, tableContent=null), ArticleFig(id=1320888964127506802, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图11, caption=菌株166明胶液化结果, figureFileSmall=B0/yh08tVXayLyzCS3hivQ==, figureFileBig=/Je9WFev0g4oElrPDevtpA==, tableContent=null), ArticleFig(id=1320888964215587187, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.12, caption=Moisture content in mouse feces, figureFileSmall=Rw+0364PkHwwHhY9cBBcOw==, figureFileBig=ERMLGbkI1vzlkulhTidVPw==, tableContent=null), ArticleFig(id=1320888964303667572, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图12, caption=小鼠粪便水分含量, figureFileSmall=Rw+0364PkHwwHhY9cBBcOw==, figureFileBig=ERMLGbkI1vzlkulhTidVPw==, tableContent=null), ArticleFig(id=1320888964408525173, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.13, caption=Organ index and fat index of mice in each group, figureFileSmall=/GhNxJhFuevIM5snU8P24A==, figureFileBig=I547KlL8u1U5nge2Py3k0w==, tableContent=null), ArticleFig(id=1320888964475634038, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图13, caption=各组小鼠脏器指数和脂肪指数, figureFileSmall=/GhNxJhFuevIM5snU8P24A==, figureFileBig=I547KlL8u1U5nge2Py3k0w==, tableContent=null), ArticleFig(id=1320888964542742903, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.14, caption=Serum lipid content in mice in each group, figureFileSmall=EGdjc7WsZFzIaBGeSt+GYA==, figureFileBig=hQtT9twGHzNEE62+c63dqw==, tableContent=null), ArticleFig(id=1320888964622434680, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图14, caption=各组小鼠血清中血脂含量, figureFileSmall=EGdjc7WsZFzIaBGeSt+GYA==, figureFileBig=hQtT9twGHzNEE62+c63dqw==, tableContent=null), ArticleFig(id=1320888964693737849, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.15, caption=Atherosclerosis index (AI) of mice in each group, figureFileSmall=MyLF7lX+mS0AhRm/hpBSag==, figureFileBig=iRk86dfZ3AV1Jryfbz5hMw==, tableContent=null), ArticleFig(id=1320888964773429626, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图15, caption=各组小鼠动脉粥样硬化指数(AI), figureFileSmall=MyLF7lX+mS0AhRm/hpBSag==, figureFileBig=iRk86dfZ3AV1Jryfbz5hMw==, tableContent=null), ArticleFig(id=1320888964848927099, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Fig.16, caption=Cholesterol content in mouse liver and feces, figureFileSmall=4PyD6N00+Pqi/3Tk7a0piw==, figureFileBig=NBahYT55D4cHShKjhewDdg==, tableContent=null), ArticleFig(id=1320888964903453052, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=图16, caption=小鼠肝脏和粪便中胆固醇含量, figureFileSmall=4PyD6N00+Pqi/3Tk7a0piw==, figureFileBig=NBahYT55D4cHShKjhewDdg==, tableContent=null), ArticleFig(id=1320888964995727741, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Table 1, caption=

Drug sensitive discs drug content and resistance judgment criteria

, figureFileSmall=null, figureFileBig=null, tableContent=
药物名称规格判定标准(抑菌圈直径:mm)
敏感(S)中介(I)耐药(R)
青霉素(PEN)10 U/片≥29−≤28
链霉素(STR)10 μg/片≥1512~17≤12
卡那霉素(KAN)30 μg/片≥1814~17≤13
万古霉素(VAN)30 μg/片≥15−≤14
四环素(TET)30 μg/片≥1512~14≤11
阿奇霉素(AZM)15 μg/片≥1814~17≤13
红霉素(ERY)15 μg/片≥2314~22≤13
诺氟沙星(NOR)30 μg/片≥1713~16≤12
), ArticleFig(id=1320888966660866430, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=表1, caption=

药敏纸片含药量及抗性判定标准

, figureFileSmall=null, figureFileBig=null, tableContent=
药物名称规格判定标准(抑菌圈直径:mm)
敏感(S)中介(I)耐药(R)
青霉素(PEN)10 U/片≥29−≤28
链霉素(STR)10 μg/片≥1512~17≤12
卡那霉素(KAN)30 μg/片≥1814~17≤13
万古霉素(VAN)30 μg/片≥15−≤14
四环素(TET)30 μg/片≥1512~14≤11
阿奇霉素(AZM)15 μg/片≥1814~17≤13
红霉素(ERY)15 μg/片≥2314~22≤13
诺氟沙星(NOR)30 μg/片≥1713~16≤12
), ArticleFig(id=1320888966757335423, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Table 2, caption=

Grouping of mice

, figureFileSmall=null, figureFileBig=null, tableContent=
试验分组小鼠数量饲养方式
空白组6只基础饲料+生理盐水
高脂组6只高脂饲料+生理盐水
阳性对照组6只高脂饲料+7 mg/kg辛伐他汀
菌液试验组6只高脂饲料+109 CFU/mL菌液
), ArticleFig(id=1320888966862193024, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=表2, caption=

小鼠分组情况

, figureFileSmall=null, figureFileBig=null, tableContent=
试验分组小鼠数量饲养方式
空白组6只基础饲料+生理盐水
高脂组6只高脂饲料+生理盐水
阳性对照组6只高脂饲料+7 mg/kg辛伐他汀
菌液试验组6只高脂饲料+109 CFU/mL菌液
), ArticleFig(id=1320888966983827841, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Table 3, caption=

Test results of drug resistance experiments

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素名称发酵粘液乳杆菌166
抑菌圈直径(mm)敏感性
青霉素(PEN)8.31±0.52R
链霉素(STR)0R
卡那霉素(KAN)0R
万古霉素(VAN)0R
四环素(TET)5.31±0.13R
阿奇霉素(AZM)7.15±0.16R
红霉素(ERY)6.08±0.21R
诺氟沙星(NOR)0R
), ArticleFig(id=1320888968921596290, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=表3, caption=

耐药性检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素名称发酵粘液乳杆菌166
抑菌圈直径(mm)敏感性
青霉素(PEN)8.31±0.52R
链霉素(STR)0R
卡那霉素(KAN)0R
万古霉素(VAN)0R
四环素(TET)5.31±0.13R
阿奇霉素(AZM)7.15±0.16R
红霉素(ERY)6.08±0.21R
诺氟沙星(NOR)0R
), ArticleFig(id=1320888969009676675, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Table 4, caption=

Body weight and feed utilization of mice in each group

, figureFileSmall=null, figureFileBig=null, tableContent=
试验分组初始体重
(g)
最终体重
(g)
体重增加量
(g)
总采食量
(g)
饲料利用率
(%)
空白组28.15±0.19a37.32±0.65c9.17±0.54d216.28±1.42b4.24±0.24b
高脂组28.41±0.47a43.92±0.61a15.51±1.28a296.45±7.24a5.23±0.34a
阳性对照组28.25±0.34a41.28±0.58b13.03±1.30c290.81±3.56a4.48±0.21b
菌液试验组27.87±0.16a42.24±1.01ab14.37±1.18b282.73±5.21a5.08±0.42a
), ArticleFig(id=1320888969101951364, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=表4, caption=

各组小鼠的体重和饲料利用率

, figureFileSmall=null, figureFileBig=null, tableContent=
试验分组初始体重
(g)
最终体重
(g)
体重增加量
(g)
总采食量
(g)
饲料利用率
(%)
空白组28.15±0.19a37.32±0.65c9.17±0.54d216.28±1.42b4.24±0.24b
高脂组28.41±0.47a43.92±0.61a15.51±1.28a296.45±7.24a5.23±0.34a
阳性对照组28.25±0.34a41.28±0.58b13.03±1.30c290.81±3.56a4.48±0.21b
菌液试验组27.87±0.16a42.24±1.01ab14.37±1.18b282.73±5.21a5.08±0.42a
), ArticleFig(id=1320888969227780485, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=EN, label=Table 5, caption=

Lee's index of mice in each group

, figureFileSmall=null, figureFileBig=null, tableContent=
试验分组Lee’s指数
空白组15.98±0.67a
高脂组16.08±0.64a
阳性对照组15.04±0.84b
菌液试验组15.13±0.34b
), ArticleFig(id=1320888969303277958, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888947337707780, language=CN, label=表5, caption=

各组小鼠Lee’s指数

, figureFileSmall=null, figureFileBig=null, tableContent=
试验分组Lee’s指数
空白组15.98±0.67a
高脂组16.08±0.64a
阳性对照组15.04±0.84b
菌液试验组15.13±0.34b
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川西高原传统牦牛酸乳中降胆固醇乳酸菌的筛选及其对高胆固醇血症小鼠的干预作用
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王思瑞 1 , 任秀梅 2 , 张诗琦 1 , 李静 1 , 王樱洁 1 , 宛祎豪 1 , 陈炼红 1, *
食品工业科技 | 生物工程 2026,47(7): 216-227
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食品工业科技 |生物工程 2026 , 47 (7) : 216 -227
川西高原传统牦牛酸乳中降胆固醇乳酸菌的筛选及其对高胆固醇血症小鼠的干预作用
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王思瑞1 , 任秀梅2, 张诗琦1, 李静1, 王樱洁1, 宛祎豪1, 陈炼红1, *
作者信息
  • 1西南民族大学药学与食品学院,四川成都 610041
  • 2四川省甘孜藏族自治州畜牧业科学研究所,四川甘孜 626000
通讯作者:
陈炼红(1967−),女,硕士,教授,研究方向:民族食品资源开发,E-mail:。
作者简介:

王思瑞(2004−),女,本科,研究方向:食品加工,E-mail:

Screening of Cholesterol-lowering Lactic Acid Bacteria from Traditional Yak Yogurt on the Western Sichuan Plateau and Its Intervention in Hypercholesterolemic Mice
Sirui WANG1 , Xiumei REN2, Shiqi ZHANG1, Jing LI1, Yingjie WANG1, Yihao WAN1, Lianhong CHEN1, *
Affiliations
  • 1School of Pharmacy and Food, Southwest Minzu University, Chengdu 610041, China
  • 2Ganzi Tibetan Autonomous Prefecture Institute of Animal Husbandry Science, Ganzi 626000, China
出版时间: 2026-04-01 doi: 10.13386/j.issn1002-0306.2025050027
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本研究旨在从川西传统发酵牦牛酸乳中筛选出一株具有高效降胆固醇作用的菌株,探究其对高胆固醇血症的干预效果。研究聚焦于川西传统发酵牦牛酸乳中分离出的295株乳酸菌,采用硫酸铁铵法和邻苯二甲醛法筛选出降胆固醇能力较高的乳酸菌,并对其进行体外益生性试验和安全性评价,通过建立高脂饮食诱导的高胆固醇血症小鼠模型评估菌株降胆固醇效果。结果表明,从295株乳酸菌中筛选得到4株胆固醇降低效率和胆固醇降低效力良好的菌株,其中菌株166在人工胃液耐受性测定试验中为4株菌株中存活率最高的,在人工肠液中培养3 h后活菌数保持在5.3×108 CFU/mL以上,具有良好的胆盐耐受性能,疏水能力和自聚集力分别达到62.3%和18.65%,经鉴定其为发酵粘液乳杆菌。此外,菌株166的有害代谢产物评价结果均为阴性,无溶血现象,不溶解明胶,对8种抗生素均表现出一定抗性,表现出良好的体外益生性和安全性。动物体内实验表明,发酵粘液乳杆菌166能显著降低小鼠血清总胆固醇(Total cholesterol,TC)、甘油三酯(Triglyceride,TG)、低密度脂蛋白胆固醇(Low-density lipoprotein cholesterol,LDLC)水平及动脉粥样硬化指数(Atherosclerosis index,AI),有效提高血清高密度脂蛋白胆固醇(High-density lipoprotein cholesterol,HDLC)含量,并有效改善小鼠Lee’s指数、肝脏指数、肾周脂肪指数和腹部脂肪指数。因此,发酵粘液乳杆菌166具有开发为功能性益生菌产品的潜力,为干预高胆固醇血症提供数据支撑。

牦牛酸乳  /  乳酸菌  /  降胆固醇  /  益生性  /  安全性  /  高胆固醇血症

This study aimed to screen a strain with high cholesterol-lowering effect from traditional fermented yak yogurt in western Sichuan province and explore its therapeutic effect on hypercholesterolemia. This study focused on 295 strains of lactic acid bacteria isolated from traditional fermented yak yogurt in Western Sichuan, and the lactic acid bacteria with high cholesterol-lowering ability were screened by ferric ammonium sulfate method and o-phthalaldehyde method, and the in vitro probiotic characterization test and safety evaluation were carried out, and the cholesterol-lowering effect of the strains was evaluated by establishing a mouse model of hypercholesterolemia induced by high-fat diet. The screening revealed four strains exhibiting favorable cholesterol-reducing efficiency and efficacy isolated from 295 lactic acid bacteria. Among the 4 strains, strain 166 had the highest survival rate in simulated gastric fluid tolerance assay test, and the number of viable bacteria remained above 5.3×108 CFU/mL after 3 h of incubation in the simulated intestinal fluid, and it possessed a good bile salt tolerance performance, with a hydrophobic capacity of up to 62.3%, auto-aggregation capacity of 18.65%, which was identified as Limosilactobacillus fermentum. In addition, strain 166 was evaluated negatively for harmful metabolites, no hemolysis phenomenon, no gelatin liquefaction, and showed some resistance to eight antibiotics, which showed good in vitro probiotic properties and safety. In vivo experiments in animals showed that Limosilactobacillus fermentum 166 could significantly reduce the serum total cholesterol, triglycerides, low-density lipoprotein cholesterol levels and atherosclerosis index of mice, effectively increase the content of serum high-density lipoprotein cholesterol, and effectively improve the Lee's index, liver index, perirenal fat index and abdominal fat index. Therefore, Limosilactobacillus fermentum 166 has the potential to be developed as a functional probiotic product, providing data support for the prevention and adjunctive therapy of hypercholesterolemia.

yak yogurt  /  lactic acid bacteria  /  cholesterol-lowering  /  probiotic properties  /  safety  /  hypercholesterolemia
王思瑞, 任秀梅, 张诗琦, 李静, 王樱洁, 宛祎豪, 陈炼红. 川西高原传统牦牛酸乳中降胆固醇乳酸菌的筛选及其对高胆固醇血症小鼠的干预作用. 食品工业科技, 2026 , 47 (7) : 216 -227 . DOI: 10.13386/j.issn1002-0306.2025050027
Sirui WANG, Xiumei REN, Shiqi ZHANG, Jing LI, Yingjie WANG, Yihao WAN, Lianhong CHEN. Screening of Cholesterol-lowering Lactic Acid Bacteria from Traditional Yak Yogurt on the Western Sichuan Plateau and Its Intervention in Hypercholesterolemic Mice[J]. Science and Technology of Food Industry, 2026 , 47 (7) : 216 -227 . DOI: 10.13386/j.issn1002-0306.2025050027
随着人们生活水平的提高和日常膳食习惯的不断转变,高脂肪饮食引起的高胆固醇血症正在影响人们的健康[1]。世界卫生组织数据表明,全球约39%的成年人总胆固醇含量超过健康水平,在中国成年居民血脂异常总体患病率达到40.4%,其中高胆固醇血症占4.9%,脂代谢紊乱问题尤为突出[2−3]。高胆固醇血症是血脂异常的一种形式,通常指空腹血浆胆固醇水平高于正常上限的异常状态,临床表现为血浆总胆固醇水平高于6.21 mmol/L[4],被认为引发心血管疾病的主要诱因之一,与多种心血管疾病的形成息息相关。当机体血清胆固醇过高时,动脉粥样硬化发生的危险也显著增加,二者存在明显的相关性[5]。还有研究表明,高胆固醇血症小鼠在中风后脑梗塞和中性粒细胞积累增加,引发先天免疫失衡[6]。对饱受心血管疾病困扰的患者来说,降低血清胆固醇水平的研究意义重大。目前,针对高胆固醇血症可采用药物治疗和日常饮食控制,虽然日常饮食中尽可能减少摄入胆固醇与脂肪较多的食物具有一定成效,但需长时间的坚持与控制,还需考虑高胆固醇血症患者的口味和偏好、他们所属的家庭、社会经济背景等多个方面,要想达到预期目的难度极大[7]。此外可以服用他汀类药物,但长期高剂量治疗,某些个体会产生不良反应,如肌肉疼痛或无力甚至引起肝功能损伤,存在副作用以及高成本的问题[8]。
益生菌被定义为对人体健康有益的活微生物,具有多种生理活性,如降低人体胆固醇、提高免疫力等[9]。益生菌可以通过促进脂肪酸氧化[10]、下调炎症信号[11]和调节肠道微生物群[12]来修复高胆固醇血症。近年来,Wang等[13]从健康人体的粪便中分离出一株益生性较好且具有降低胆固醇作用的加氏乳杆菌TF08-1;赵丽娜等[14]在新疆赛里木酸奶中筛选出了3株高降解胆固醇的植物乳杆菌,并证实其具有一定的安全性。牦牛乳作为川西游牧民族的主要传统乳制品,其乳脂含量较普通牛乳更高,在胃肠道消化的后期表现出更强的脂肪分解能力,比牛乳脂释放更多的不饱和脂肪酸[15],从中筛选出胆固醇降低能力较强的乳酸菌菌株用于高脂饮食小鼠的影响研究具有可行性,目前还未见有对川西传统发酵牦牛酸乳源降胆固醇微生物的系统研究,本研究既加大了对川西传统发酵牦牛酸乳中微生物资源的开发,又为区域特色发酵乳品功能化转型奠定了基础。
本研究聚焦于川西高原传统牦牛酸乳,通过硫酸铁铵法和邻苯二甲醛法筛选出具有高效降胆固醇能力的乳酸菌,并通过高脂饮食诱导的高胆固醇血症小鼠模型,探究该菌株的降胆固醇功效,以期为未来川西高原传统牦牛酸乳源益生菌开发为辅助干预高胆固醇血症提供实验依据。
6周龄雄性昆明小鼠 成都达硕实验动物有限公司(许可证号SCXK(川)2022-0039),实验经西南民族大学实验动物伦理委员会批准(批准号SMU-202501124);试验菌株 从川西7个县15个地区传统发酵牦牛酸乳中分离鉴定出的295株乳酸菌(乳杆菌属、乳球菌属、肠球菌属);金黄色葡萄球菌ATCC6538、MRS琼脂培养基、MH琼脂培养基、LB培养基、赖氨酸、精氨酸和鸟氨酸脱羧酶生化管 青岛海博生物技术有限公司;哥伦比亚血琼脂培养基 梅里埃(上海)生物制品有限公司;邻苯二甲醛、硫酸铁铵、牛胆盐 成都康迪生物技术有限公司;胆固醇 分析纯,Solarbio公司;胃蛋白酶(酶活力1:3000)、胰蛋白酶(酶活力1:250) 分析纯,如吉生物科技;药敏片、无菌液体石蜡 金澳科技化工有限公司;辛伐他汀 成都恒瑞制药有限公司;总胆固醇测定试剂盒、血清甘油三酯试剂盒、血清高密度脂蛋白试剂盒、血清低密度脂蛋白试剂盒 南京建成生物工程研究所;普通饲料、高脂饲料 北京科澳协力饲料有限公司。
5804R型冷冻离心机 德国Eppendorf公司;ZWY-100H型恒温培养振荡器、MLS-3020高压灭菌锅 上海智城分析仪器制造有限公司;MP512-02型精密pH计 上海三信仪表厂;UV-6100分光光度计 上海美普达公司;Rayto RT-6100酶标仪 济南骏驰生物科技有限公司。
将试验菌株以4%的接种量分别接种于MRS肉汤培养基中,充分混匀后置于37 ℃、180 r/min恒温培养箱中振荡培养,经24 h活化培养后,通过传代获得生长状况良好的菌悬液[16]。
配制梯度胆固醇标准品,采用硫酸铁铵法(GB 5009.128-2016),分别检测295株待测菌株降胆固醇能力。在560 nm波长处测定吸光值,绘制胆固醇标准曲线[17],吸光度值(y)对胆固醇浓度(x)的标准曲线为y=3.035x−0.0032,R2=0.9985。
参考李尧[18]的方法及实验室前期摸索得出,将活化后的295株菌接种于等量含胆固醇的MRS肉汤培养基(胆固醇浓度为1 mg/mL),37 ℃培养48 h左右至稳定期,各培养液的浊度基本保持一致,测定OD600值在0.8~0.9范围。3000 r/min离心10 min后,取1 mL上清液于离心管,加入4倍体积无水乙醇,充分摇匀,再次离心10 min,取二次离心上清液与2 mL硫酸铁铵显色剂充分涡旋混合,于室温时560 nm处测定吸光值,通过胆固醇标准曲线计算发酵后上清液中胆固醇含量,以未接菌的含胆固醇MRS培养基为对照测定其中的胆固醇总量,降低率计算如下式:
$ {\mathrm{W}}({\text{%}}) = \frac{{\mathrm{A}}-{\mathrm{B}}}{{\mathrm{A}}}\times 100 $
式中:W—胆固醇降低率,%;A—接种前培养基胆固醇含量,μg;B—发酵后上清液中胆固醇含量,μg。
制备系列浓度胆固醇标准溶液,采用邻苯二甲醛法[19],于560 nm测定吸光值,绘制标准曲线,吸光度值(y)对胆固醇浓度(x)的标准曲线为y=3.917x+0.025,R2=0.9994。
将活化后的试验菌株分别接种于MRS-CHOL肉汤培养基(MRS培养基中,含0.2%(m/v)巯基乙酸钠,0.2%(m/v)胆盐,100 μg/mL的胆固醇),于37 ℃培养24 h,分别取3 mL培养液,用等体积生理盐水洗涤两次后,测定培养物在600 nm的吸光度值,该值即为乳酸菌生长量。再取1 mL培养液在4 ℃、10000 r/min离心6 min,取上清液采用邻苯二甲醛法测定各菌株降低胆固醇的量,再结合乳酸菌生长量的测定结果,可知各菌株的胆固醇降低效力[20]。降低效力计算如下式:
$ {\mathrm{Z}}=\frac{{\mathrm{A}}-{\mathrm{B}}}{{\mathrm{D}}} $
式中:Z—胆固醇降低效力,μg/u;A—接种前培养基中胆固醇含量,μg;B—发酵后上清液中胆固醇含量,μg;D—乳酸菌生长量,定义为测定乳酸菌降低效力的培养液的1单位浊度值(即1 OD600)表示。
参考汤凯等[21]的方法,用蒸馏水稀释2 mL 1 mol/L的盐酸,调节pH分别至1.5、2.5、3.5、4.5、5.5,以1 g/100 mL的比例加入胃蛋白酶,用0.22 μm的微孔滤膜除菌。将活化后的菌液以10%体积分数接种于处理好的人工胃液中,置于37 ℃、100 r/min恒温摇床培养,在0、1、2、3 h通过涂布稀释法测定样品中的活菌数,根据下式计算存活率:
$ 存活率(\text{%})=\frac{{{\mathrm{N}}}_{1}}{{{\mathrm{N}}}_{0}}\times \text{100} $
式中:N0为0 h测定的活菌数,CFU/mL;N1为3 h测定的活菌数,CFU/mL。
参考肖秋颖等[22]的方法,取3.4 g KH2PO4加入蒸馏水250 mL,调节pH至7.6,以1 g/100 mL的比例加入胰蛋白酶,用0.22 μm的微孔滤膜除菌。将活化后的菌液以10%的接种量接种于处理好的人工肠液中,混匀后在37 ℃、100 r/min恒温摇床进行培养,在0、1、2、3、4 h通过涂布稀释法测定样品中的活菌数。根据下式计算存活率:
$ 存活率(\text{%})=\frac{{{\mathrm{N}}}_{\text{2}}}{{{\mathrm{N}}}_{0}}\times \text{100} $
式中:N0为0 h测定的活菌数,CFU/mL;N2为肠液处理后测定的活菌数,CFU/mL。
参考檀茜倩等[23]的方法加以改进,制备含牛胆盐梯度浓度为0、0.03、0.08、0.13、0.18、0.23、0.28和0.33 g/100 mL的MRS肉汤培养基,将活化后的菌液以10%体积比接种到该培养基中,37 ℃恒温培养24 h,采用涂布稀释法进行活菌计数。
将试验菌株接种于液体MRS培养基,37 ℃培养18 h,5000 r/min离心10 min,用0.1 mol/L KNO3(pH6.2)洗涤两次后,用该溶液悬浮菌液,在600 nm处调整其初始吸光度至OD600=0.5±0.02。再吸取菌悬液3 mL,加入l mL二甲苯,室温预培养10 min,涡流快速混合2 min,室温静止放置15 min使之分层,吸取下层水相,以缓冲液为空白对照,在600 nm测量最终吸光度A。根据下式进行疏水能力计算:
$ 疏水性(\text{%})=\left(1-\frac{{\mathrm{A}}}{0.5}\right)\times 100 $
参考申辉[24]的方法加以修改,试验菌株活化后经5000 r/min离心10 min,弃上清并用PBS缓冲液重悬冲洗3次后离心得到菌体。再次重悬并将其在600 nm下的吸光度调到OD600=0.5±0.02。取2 mL菌悬液充分振荡混匀15 s,并于37 ℃培养2 h。吸取上层菌液1 mL在600 nm下测定吸光值A。根据下式进行自聚集能力计算:
$ 自聚集能力(\text{%})=\left(1-\frac{{\mathrm{A}}}{0.5}\right)\times 100 $
参照郑淼等[25]的方法作适当修改,将活化好的菌株接种到蛋白胨水培养基,于37 ℃培养箱中培养48~72 h后,加入1 mL乙醚,充分振荡,静置5 min后,使乙醚层浮于培养物的液面。再沿管壁缓慢加入1 mL的吲哚试剂,观察试验结果,同时做空白对照。滴加吲哚试剂后在两层液体交界处出现红色环者为阳性,不变色的为阴性。
参考Motey等[26]的方法,将活化好的菌株接种于哥伦比亚血琼脂平板,以金黄色葡萄球菌(ATCC25923)作为阳性对照,倒置培养48 h,观察菌落是否出现溶血现象。
按照氨基脱羧酶生化管使用要求,将试验菌分别接种于生化管中,置于培养箱中37 ℃培养48 h左右。通过生化管颜色变化特征,对菌株的氨基脱羧酶活性进行判定。
试验菌株活化后调整菌体浊度为0.5麦氏单位,均匀涂布于MH琼脂培养基表面,将药敏纸片贴于培养基表面。培养24 h后测量抑菌圈直径,标准如表1所示[27]。
将试验菌株进行明胶液化试验[28],以无菌生理盐水作为阴性对照,通过观察试管内明胶的融化状况来判断实验结果。若出现明胶融化现象,则判定为明胶液化反应阳性。
对试验小鼠进行为期5 d的适应性饲养后,按表2将其随机分为4组,试验进行4周。
菌剂制备:将发酵粘液乳杆菌166活化后,调整菌液浓度至109 CFU/mL,每天现配现用。
辛伐他汀溶液:将辛伐他汀药物按7 mg/kg溶于生理盐水中,置于室温保存备用。
每天定时进行灌胃,灌胃量按体重以0.1 mL/10 g计算,连续灌胃4周。
按时记录小鼠采食量(g)及体重(g),根据下式计算饲料利用率[29]。
$ 饲料利用率(\text{%})=\frac{体重增加量}{采食量}\times 100 $
将每周采集(4周)的新鲜粪便收集于铝盒中,105 ℃干燥48 h至恒重,根据下式计算粪便含水量[30]。
$ 粪便含水量(\text{%})=\frac{{{\mathrm{W}}}_{1-}{{\mathrm{W}}}_{2}}{{{\mathrm{W}}}_{1}-{{\mathrm{W}}}_{0}}\times 100 $
式中:W0—铝盒重量,g;W1—新鲜粪便及铝盒重量,g;W2—恒重粪便及铝盒重量,g。
记录各组小鼠灌胃4周后的体重并测量体长。根据下式计算Lee’s指数[31]。
$ {{\mathrm{Lee{\text’} s }}}指数=\sqrt[{3}]{\frac{体重({\mathrm{g}})\times 1000}{体长({\mathrm{cm}})}} $
试验第4周,小鼠禁食12 h后,将小鼠称重后心脏采血后致死,分离获取其心脏、肾脏、脾脏、肝脏、腹部脂肪和肾周脂肪,并准确称取各部分质量,按下式计算脏器指数和脂肪指数[32]。
$ \begin{split} & 脏器指数=\frac{各脏器重量(\mathrm{mg})}{小鼠体重(\mathrm{g})} \\ &脂肪指数=\frac{脂肪重量(\mathrm{mg})}{小鼠体重(\mathrm{g})}\end{split} $
灌胃4周后将全部小鼠禁食12 h,麻醉后摘眼球取血,血液经2000 r/min离心10 min,取上清备用。按照试剂盒说明,测定小鼠血清TC、TG、HDLC以及LDLC含量,并按下式计算AI值[33]。
$ 动脉粥样硬化指数({\mathrm{AI}})=\frac{{\mathrm{TC}}-{\mathrm{HDLC}}}{{\mathrm{HDLC}}} $
小鼠处死后,采集肠道新鲜粪便及肝脏组织样本,保存于−80 ℃。按胆固醇测定试剂盒说明书分别检测肝脏与粪便胆固醇含量。
所有数据均为3次平行实验所得的平均值,数据以平均值±标准差的形式表示,采用SPSS 26进行数据统计方差及显著性分析(P<0.05),采用Origin 2021绘图。
基于硫酸铁铵法评估295株乳酸菌的降解胆固醇能力,统计显示菌株间降胆固醇能力存在差异,胆固醇降低率范围为0.30%~58.00%。由图1可知,降低率在30%~40%的菌株数量最多,有107株,有18株乳酸菌胆固醇降低率在10%以下,有118株乳酸菌的降低率在30%以下,有53株乳酸菌降低率在40%~50%,有17株乳酸菌降低率超50%,其中6株胆固醇降低率在54%以上,分别是菌株114(54.33%)、菌株121(56.53%)、菌株135(54.15%)、菌株160(56.96%)、菌株166(58.00%)和菌株227(56.47%),它们显示出了较强的降胆固醇的能力。后续对这6株乳酸菌进行胆固醇降低效力研究。
对6株乳酸菌的胆固醇降低率和降低效力进行综合分析,如图2可知,菌株121、160、166和227的胆固醇降低率及胆固醇降低效力显著高于菌株114、135(P<0.05),因此,选择菌株121、160、166和227进行后续益生功能的研究。
由图3所示,总体上4株待测菌株的活菌数都随着培养时间的延长呈递下降趋势,表明人工胃液对乳酸菌的生长活性具有抑制效应。菌株121和菌株160在pH1.5的人工胃液中培养3 h后活菌数为0,说明这两株菌都不能抵御人工胃液的酸性环境,在强酸环境中不能够存活。菌株227在pH1.5的人工胃液中培养3 h后活菌数依然保持在很高的程度,并且在不同pH的人工胃液中活菌数均稳定在2.4×108 CFU/mL以上,说明其展现了较好的人工胃液耐受性,能够抵御极端的酸性环境,值得进一步研究。菌株166在pH4.5的人工胃液中活菌数最高,达到了5.7×108 CFU/mL。而在pH5.5的人工胃液中,活菌数稍有下降,但总体上保持较高的活菌数,并且没有大范围的菌株死亡现象,下降趋势较为平缓。与菌株227相比,菌株166对人工胃液的耐受性能表现更为优异,为其在人体发挥相应的益生功能提供了数据支撑。
由图4可知,四株菌都出现了随着pH升高,菌株存活率先上升后下降的趋势,这与宛祎豪等[34]和黄晓英[35]的研究结果一致,其使用的菌株都源自牦牛酸奶,这可能是由于牦牛酸乳发酵过程中,pH变化复杂,菌株在长期适应这种特殊环境后,形成了独特的耐酸特性。菌株166在经3 h不同pH环境处理后,存活率均超过20%,最高达到50.44%,存活率显著高于菌株121、160和227(P<0.05),且在不同pH条件下变化曲线平稳、舒缓,展现出较好的胃酸耐受特性,具有实际应用潜力。菌株227在不同pH的胃部环境下,存活率始终维持在15%~30%的范围内,存活率显著高于菌株121和160(P<0.05),在各pH环境中变化平稳。因此,选取菌株166和菌株227进行后续耐受能力测定。
肠道微环境对益生菌群落的影响要素主要包括共生微生物区系、碱性环境(pH≈7.6)、多种蛋白水解酶及胆汁酸复合物,根据文献报道,食物在小肠内的停留时间通常为1~4 h[36],为确保乳酸菌能够发挥益生功效,维持足够的活菌数是关键前提,同时还要具备对肠液环境的适应能力。由图5可知,随着培养时间的延长,菌株166和菌株227在人工肠液中的活菌数呈现出逐步下降的趋势,但都未出现大批死亡现象。在培养3 h后两株菌的活菌数依然保持在3.8×108 CFU/mL以上;在培养4 h后,虽然菌株166依然有下降的趋势,但其活菌数仍然显著高于菌株227(P<0.05),其存活率为35.81%,说明两菌株都具有良好的肠液耐受性能,但菌株166对人工肠液的耐受力稍高于菌株227。
胆盐作为胆汁发挥消化与吸收功能的核心成分,由肝细胞产生的胆汁酸与甘氨酸或牛磺酸结合而成,在人体小肠环境中,胆盐浓度呈现动态变化,通常维持在0.03~0.3 g/100 mL的区间范围内[22]。由图6可知,随着胆盐溶液浓度的逐步升高,菌株166与菌株227的活菌数均呈递减态势。当两株菌处于低胆盐浓度时,活菌数的下降幅度相对平缓;但随着胆盐浓度增加,菌株出现了大规模的死亡现象,活菌数急剧下降。当胆盐浓度达到0.33 g/100 mL时,菌株166、227的活菌数分别降至最低点,为1.9×108 CFU/mL和2.5×108 CFU/mL。在整个实验过程中,菌株227的活菌数始终优于菌株166,但两者均展现出良好的胆盐耐受性能,这为其在肠道复杂生态环境中实现定植增殖提供了有力的数据支撑。
乳酸菌表面的疏水性越高,其黏附性就越强。当乳酸菌的黏附性增强时,能有效利用营养物质,促进自身的生长繁殖[37]。近年来,细菌表面疏水性常被用作评估细菌的黏附能力的关键指标。从图7中可以看出,菌株166和菌株227都具有良好的疏水能力,分别为62.3%和54.4%,相比而言,菌株166疏水性优于菌株227。
自聚集能力是益生菌发挥益生功效的关键特征,不仅反映细菌黏附能力,也影响着其在肠道的定植[33,38]。从图中可以看出,菌株166和菌株227的自聚集能力分别为18.65%和17.08%,对比而言,菌株166自聚集能力更强。综合菌株胆固醇降低率和降低效力及不同益生特性,选择菌株166作为目标菌株进行下一步试验,菌株166经实验室前期鉴定为发酵粘液乳杆菌。
有研究表明,部分乳酸菌在发酵过程会产生有害代谢产物,如吲哚、溶血素等,严重危害人体健康[39]。菌株166的吲哚试验结果如图8所示,加入吲哚试剂后菌株166的液面颜色未发生变化,与空白对照组液面颜色一致,都未出现红色,判定为阴性结果。由此说明,发酵粘液乳杆菌166在生长过程中不会代谢产生色氨酸酶,不能催化色氨酸分解为吲哚。
在某些特定条件下,部分乳酸菌菌株可能会表现出溶血性,这不仅其对血液系统的潜在危害,严重时可能诱发败血症,因此开展乳酸菌的溶血特性评估实验具有重要意义[40]。溶血试验结果如图9所示,阳性对照菌菌落周围出现了溶血环,为β-溶血,而发酵粘液乳杆菌166正常生长,菌落周围无透明圈出现,为不溶血。
某些乳酸菌可以分泌氨基酸脱羧酶,催化氨基酸的脱羧反应,进而生成相应的生物胺类物质,而生物胺是致癌性N-亚硝基化合物的前体物质,对人体健康有着不良影响[41−42]。因此对乳酸菌进行氨基酸脱羧酶活性分析十分必要。氨基酸脱羧酶活性试验如图10所示,加入菌株166的三种氨基酸生化管均变为黄色,该菌氨基酸脱羧酶反应为阴性,说明发酵粘液乳杆菌166不产生氨基酸脱羧酶,不会造成生物胺积累。
如表3所示,青霉素、四环素、阿奇霉素和红霉素对发酵粘液乳杆菌166产生了抑菌圈,但根据抗性判定标准其表现为抗性,对链霉素、卡那霉素、万古霉素和诺氟沙星不产生抑菌圈,同样表现为抗性。菌株166对8种抗生素表现为耐药,在发酵粘液乳杆菌中虽不常见,但值得强调的是,耐药性不等同于致病性。未来应用中将通过耐药基因筛查确保菌株166使用安全。
研究表明,具备明胶液化特性的菌株通常具有致病潜能[43],为防止可能的安全隐患,对菌株166进行了明胶液化的评价。结果如图11所示,菌株166与空白组结果一致,均未使明胶固体培养基表面发生液化,进一步证明了菌株166的安全性。
试验期间,小鼠均保持正常生理状态,未观察到小鼠出现精神萎靡、食欲不振等异常情况,说明灌胃发酵粘液乳杆菌166不会对小鼠产生毒副作用。如表4所示,各组试验小鼠的体重增加量介于9.17~15.51 g区间,总摄食量处于216.28~296.45 g范围,饲料利用率维持在4.24%~5.23%之间。饲喂高脂饲料对小鼠体重增长具有显著促进作用(P<0.05),高脂组小鼠最终体重为四组中最高。菌液试验组小鼠体重虽较高脂组有所下降,但仍高于阳性对照组,且差异都不显著(P>0.05),但其体重增加量显著低于高脂组小鼠(P<0.05)。阳性对照组小鼠最终体重和体重增加量均显著低于高脂组小鼠(P<0.05)。表明辛伐他汀类药物和菌株166都能有效降低高脂小鼠体重,与Park等[44]的结果类似。在饲料利用率方面,高脂组小鼠饲料利用率为四组中最高,菌液试验组的饲料利用率稍低于高脂组,但不存在显著性差异(P>0.05),并且菌液试验组饲料利用率达到了5.08%,显著高于空白组和阳性对照组(P<0.05),说明灌胃菌液可能通过改善小鼠对饲料的消化吸收,进而提高饲料利用率。综合表明,发酵粘液乳杆菌166能够提高小鼠食欲,促进脂肪代谢,防止脂肪过度存积。灌喂菌株166在小鼠体内发挥了一定的作用。
灌胃试验期间,小鼠粪便水分含量变化如图12所示。空白组小鼠粪便含水量保持相对稳定,无明显变化,在37.52%~39.83%之间浮动。高脂组小鼠粪便含水量急剧下降,由初始含量34.83%的含水量降至最终的19.41%,表明小鼠摄取高脂饲料会导致其粪便干燥,水分减少;阳性对照组小鼠粪便中含水量整体呈下降趋势,但在第21 d时有所回升,其含量始终高于高脂组,试验结束时也较高脂组增加了7.53%,表明他汀类药物能够在一定程度上增加高脂饮食小鼠粪便中的水分含量,从而缓解因高脂饮食引起的粪便干燥问题;菌液试验组小鼠粪便含水量变化很小,基本保持稳定,且从第7 d开始明显高于高脂组和阳性对照组,试验结束时也较高脂组和阳性对照组分别增加了17.80%和10.27%,具有显著性差异(P<0.05),与空白组小鼠粪便含水量无显著差异(P>0.05),效果最佳。综上所述,菌株166处理能够改善高脂膳食所致的小鼠粪便干燥问题,并有效提升小鼠粪便含水量,进而改善高胆固醇血症小鼠的整体健康状态。
Lee’s指数通过衡量体重与体长的比率,评估机体肥胖程度[45]。由表5所示,菌液试验组的Lee’s指数虽与阳性对照组无显著差异(P>0.05),但显著低于高脂组与空白组(P<0.05),表明发酵粘液乳杆菌166可能通过调节能量代谢稳态对小鼠肥胖表型产生显著改善作用[46]。
如图13所示,四组小鼠的心脏、脾脏以及肾脏指数均处于较为接近的水平,没有显著性差异(P>0.05),表明灌胃菌株166不会对小鼠造成明显的毒副作用,进一步证明了发酵粘液乳杆菌166的安全性。分析肝脏指数发现,高脂组呈现最高数值,阳性对照组小鼠的肝脏指数低于高脂组小鼠,但差异不显著(P>0.05),而菌液试验组小鼠的肝脏指数下降至37.80 mg/g,不仅显著低于高脂组小鼠的肝脏指数(P<0.05),且较阳性对照组有进一步改善作用,与空白组无显著差异(P>0.05),表明菌株166在降低高胆固醇血症小鼠肝脏胆固醇积累方面具有显著效果。
分析脂肪指数发现,高脂组小鼠的腹部脂肪指数和肾周脂肪指数显著高于空白对照组(P<0.05),并且都为四组中最高,分别达到了31.24 mg/g和9.86 mg/g。菌液试验组小鼠腹部脂肪指数和肾周脂肪指数相较于阳性对照组及高脂组均呈下降趋势,但差异不显著(P>0.05)。结果表明,灌胃菌株166能降低高脂小鼠的肥胖程度,减少肝脏、腹部及肾周脂肪的积累,降低心血管疾病风险,有效改善小鼠健康状况。
由图14可知,高脂组小鼠血清TG和TC含量分别为2.56 mmol/L和4.42 mmol/L,显著高于空白组小鼠TG和TC含量(P<0.05);菌液试验组的小鼠的血清TG、TC含量分别为1.57和3.08 mmol/L,显著低于高脂组(P<0.05);而阳性对照组的血清TC含量为2.8 mmol/L,显著低于菌液试验组(P<0.05),TG含量为1.42 mmol/L稍低于菌液试验组,差异不显著(P>0.05)。说明灌胃菌液166可以降低血清中TG和TC的含量。
高脂组小鼠血清中HDLC含量为0.98 mmol/L,显著低于其他三组(P<0.05),菌液试验组含量为1.8 mmol/L稍高于阳性对照组,但没有显著差异(P>0.05),这两组的血清HDLC含量与空白组相比,仍维持在较高水平,且存在显著性差异(P<0.05),证明其对升高血清中HDLC含量具有一定积极作用;高脂组小鼠血清中的LDLC含量最高,为1.6 mmol/L,显著高于空白组小鼠(P<0.05),菌液试验组和阳性对照组LDLC含量分别为0.74和0.68 mmol/L,都低于高脂组,并且相差很大,存在显著性(P<0.05),而菌液试验组LDLC含量稍高于阳性试验组,差异不明显。说明灌胃菌液166虽然没有使血清中HDLC含量和LDLC含量达到正常水平,但仍然有一定的影响作用,这可能与试验周期短有一定关系。
如图15所示,高脂组小鼠的AI值最大,达到了3.51,其次是菌液试验组,AI值为0.71,显著低于高脂组小鼠的AI值(P<0.05),并且与阳性对照组不存在显著性差异。可以得出结论,发酵粘液乳杆菌166可以降低血清中LDLC、TC、TG的含量,同时增加HDLC的含量,在动脉粥样硬化疾病预防方面表现出显著优势,对改善高胆固醇血症小鼠的健康状况具有一定价值意义。
由图16可知,高脂组小鼠由于摄入大量高脂饲料,其肝脏组织中胆固醇水平上升至1.19 mmol/L,显著高于其他三组(P<0.05),表明高脂饮食模式会造成小鼠体内脂质代谢失衡,过多量的胆固醇在肝脏部位异常积聚。菌液试验组和阳性对照组肝脏中胆固醇含量分别为0.39、0.47 mmol/L,与高脂组相比差异显著(P<0.05),且菌液试验组稍低于阳性对照组。该结果与丁淑娟[47]的研究结果一致。证明他汀药物和菌株166都可以减少肝脏中胆固醇的积存,对小鼠肝脏胆固醇水平的降低具有显著的干预效果。
高脂组小鼠粪便中胆固醇水平较空白组显著升高(P<0.05),达到0.33 mmol/L,阳性对照组粪便胆固醇水平为0.37 mmol/L,虽略高于高脂组但无显著性差异(P>0.05),显著高于空白组的0.20 mmol/L(P<0.05);菌液试验组小鼠粪便胆固醇水平高达0.43 mmol/L,显著高于空白组、高脂组和阳性对照组(P<0.05)。表明菌株166干预后能有效减少高胆固醇血症小鼠胆固醇在肝脏中的累积,促进胆固醇经肠道排泄,相较于阳性对照药物辛伐他汀,菌株166展现出更优的降胆固醇效果,其在降低胆固醇方面具有显著优势,这与万婧倞[48]的研究结果一致。
本研究从川西传统发酵牦牛酸乳中分离鉴定出的295株乳酸菌中筛选得到一株降胆固醇能力较强且具有良好益生性和安全性的菌株166,其胆固醇降低率达到58.00%,在当前关于传统发酵牦牛酸乳中降胆固醇乳酸菌的研究方面,菌株166的胆固醇降低率和降低效力都处于较高水平,具有一定的应用潜力。经体外试验分析菌株166具备良好益生性和安全性,动物体内实验表明,发酵粘液乳杆菌166能显著降低小鼠血清TC、TG、LDLC含量及AI值,有效提高HDLC含量,并有效改善小鼠Lee’s指数、肝脏指数、肾周脂肪指数和腹部脂肪指数。本研究表明菌株166对高胆固醇血症小鼠模型具有良好的益生性能,研究结果不仅丰富了我国乳酸菌资源,还为干预高胆固醇血症提供了新的研究方向。

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2026年第47卷第7期
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doi: 10.13386/j.issn1002-0306.2025050027
  • 接收时间:2025-05-08
  • 首发时间:2026-10-24
  • 出版时间:2026-04-01
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  • 收稿日期:2025-05-08
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    1西南民族大学药学与食品学院,四川成都 610041
    2四川省甘孜藏族自治州畜牧业科学研究所,四川甘孜 626000

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陈炼红(1967−),女,硕士,教授,研究方向:民族食品资源开发,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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