Article(id=1320890091975852725, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320889471592149951, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025020073, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1739289600000, receivedDateStr=2025-02-12, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792854338877, onlineDateStr=2026-10-24, pubDate=1771084800000, pubDateStr=2026-02-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792854338877, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792854338877, creator=13701087609, updateTime=1792854338877, updator=13701087609, issue=Issue{id=1320889471592149951, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='4', pageStart='1', pageEnd='524', issueExtLink='null', onlineDate='null', pubDate='1771084800000', pubDateStr='2026-02-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792854190967, creator='13701087609', updateTime=1792854190967, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=435, endPage=442, ext={EN=ArticleExt(id=1320890092210733750, articleId=1320890091975852725, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Relieving Effect of Heyndrickxia coagulans BC07 on Loperamide Hydrochloride-induced Constipation in Mice, columnId=1261343846965830077, journalTitle=Science and Technology of Food Industry, columnName=Nutrition and Healthcare, runingTitle=null, highlight=null, articleAbstract=

Objective: To investigate the effect of Heyndrickxia coagulans BC07 on loperamide hydrochloride-induced constipation in mice. Methods: Thirty mice were randomly divided into three groups: the normal control group (NC), the model group (MC) and the BC07 group, with 10 mice in each group. The NC and MC groups were intragastrically administered normal saline, while the BC07 group was intragastrically given Heyndrickxia coagulans BC07 bacterial solution for 14 days. From the 15th to the 17th day, loperamide hydrochloride was used to establish the constipation model. The time to the first black feces and intestinal transit in mice were observed. Feces, serum, cecum, and colon samples of mice were collected. Results: The intake of Heyndrickxia coagulans BC07 could significantly increase the feces water content in constipated mice (P<0.05), shorten the time to the first black feces in mice (P<0.05), and promote the propulsion rate of the small intestine. Meanwhile, it could effectively regulate the concentrations of motilin (MTL), vasoactive intestinal peptide (VIP), substance P (SP), and somatostatin (SS) in constipated mice. It also reduced the contents of pro-inflammatory cytokines IL-6 and TNF-α in the mouse serum and increased the level of the anti-inflammatory factor IL-10. In addition, after the supplementation of Heyndrickxia coagulans BC07, the structure of the intestinal microbiota in constipated mice could be effectively regulated, increasing the relative abundance of beneficial microbiota in the mouse intestine and significantly decreasing the relative abundance of harmful microflora (P<0.05). Conclusion: Heyndrickxia coagulans BC07 has the effect of increasing fecal water content and alleviating defecation difficulties. It is speculated that it may relieve constipation by regulating gastrointestinal peptide transmitters, improving the structure of the intestinal microbiota, and promoting intestinal peristalsis.

, authors=Sufen KONG, Mingming ZHU, Jianguo ZHU, Shuguang FANG*, authorsList=Sufen KONG, Mingming ZHU, Jianguo ZHU, Shuguang FANG, authorCompany=null, correspAuthors=Shuguang FANG, 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=1320890095541011143, articleId=1320890091975852725, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=凝结海恩德里克斯氏菌BC07对盐酸洛哌丁胺诱导小鼠便秘的缓解作用, columnId=1261343850979779009, journalTitle=食品工业科技, columnName=营养与保健, runingTitle=null, highlight=null, articleAbstract=

目的:研究凝结海恩德里克斯氏菌BC07对盐酸洛哌丁胺诱导小鼠便秘的影响。方法:将30只小鼠随机分为对照组(NC)、模型组(MC)、BC07组,每组10只,NC组和MC组灌胃生理盐水,BC07组灌胃凝结海恩德里克斯氏菌BC07菌液,持续14 d。第15~17 d使用盐酸洛哌丁胺造模,观察小鼠首粒黑便排出时间及肠转运情况,收集小鼠粪便,血清,盲肠和结肠样本。结果:摄入凝结海恩德里克斯氏菌BC07能够显著增加便秘小鼠的粪便水分含量(P<0.05),缩短小鼠首粒黑便排出时间(P<0.05),促进小肠的推进率;同时还能有效调节便秘小鼠胃动素(Motilin,MTL)、血管活性肠肽(Vasoactive intestinal peptide,VIP)、P物质(Substance P,SP)、生长抑素(Somatostatin,SS)的浓度,并能降低小鼠血清中细胞促炎因子IL-6和TNF-α的含量,提高抗炎因子IL-10的水平;此外,补充凝结海恩德里克斯氏菌BC07后,能有效调节便秘小鼠的肠道菌群结构,增加小鼠肠道有益菌群的相对丰度,显著降低有害菌群的相对丰度(P<0.05)。结论:凝结海恩德里克斯氏菌BC07具有提高粪便水分含量、减轻排便困难的作用,推测可能通过调节胃肠肽递质、改善肠道菌群结构、促进肠道蠕动来缓解便秘。

, authors=孔素芬, 朱明明, 朱建国, 方曙光*, authorsList=孔素芬, 朱明明, 朱建国, 方曙光, authorCompany=null, correspAuthors=方曙光, authorNote=

孔素芬(1984−),女,本科,中级工程师,研究方向:益生菌菌种及功能开发研究,E-mail:

, correspAuthorsNote=
方曙光(1974−),男,博士,教授,研究方向:益生菌功能性、产业化关键技术开发,E-mail:。
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孔素芬(1984−),女,本科,中级工程师,研究方向:益生菌菌种及功能开发研究,E-mail:

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孔素芬(1984−),女,本科,中级工程师,研究方向:益生菌菌种及功能开发研究,E-mail:

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Journal of Medical Microbiology, 2016, 65(10): 1049−1059., articleTitle=null, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1320890096002384584, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, xref=null, ext=[AuthorCompanyExt(id=1320890096010773193, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, companyId=1320890096002384584, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Wuhan Wecare Probiotic Research Institute Co., Ltd., Wuhan 430000, China), AuthorCompanyExt(id=1320890096023356106, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, companyId=1320890096002384584, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=武汉微康益生菌研究院有限公司,湖北武汉 430000)])], figs=[ArticleFig(id=1320890101048132325, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.1, caption=Fecal particle number and water content of mice (n=8), figureFileSmall=xfjJ1CaXfeCmiPKwiULPyQ==, figureFileBig=lCq8bWBtVl4nLYjI9hRREw==, tableContent=null), ArticleFig(id=1320890102960734950, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图1, caption=小鼠粪便颗粒数和含水量(n=8)

注:图中不同小写字母表示有显著差异(P<0.05),图2、图4~图8同。

, figureFileSmall=xfjJ1CaXfeCmiPKwiULPyQ==, figureFileBig=lCq8bWBtVl4nLYjI9hRREw==, tableContent=null), ArticleFig(id=1320890103313056487, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.2, caption=Small intestinal motility of mice in each group (n=8), figureFileSmall=Jdsd4qYqgcTqWtABvP7h+g==, figureFileBig=s77HME62CCxxaK3CAMrfVQ==, tableContent=null), ArticleFig(id=1320890103464051432, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图2, caption=各组小鼠小肠运动能力(n=8), figureFileSmall=Jdsd4qYqgcTqWtABvP7h+g==, figureFileBig=s77HME62CCxxaK3CAMrfVQ==, tableContent=null), ArticleFig(id=1320890103526965993, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.3, caption=Observation of colon tissue, figureFileSmall=tXmrtrpMrO5giPXfGROj8w==, figureFileBig=VbcvdM94ZBMDDMl136tmPA==, tableContent=null), ArticleFig(id=1320890103598269162, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图3, caption=结肠组织观察

注:各组小鼠结肠切片H&E染色(150×)。

, figureFileSmall=tXmrtrpMrO5giPXfGROj8w==, figureFileBig=VbcvdM94ZBMDDMl136tmPA==, tableContent=null), ArticleFig(id=1320890103682155243, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.4, caption=Effects of BC07 on constipation-related GI neurotransmitters (n=5), figureFileSmall=lpMbuHey+SNdeu1Q7g7JcQ==, figureFileBig=MJsJeGiTFO2wYFEFoJB7qg==, tableContent=null), ArticleFig(id=1320890103761847021, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图4, caption=BC07对便秘相关胃肠道神经递质的影响(n=5), figureFileSmall=lpMbuHey+SNdeu1Q7g7JcQ==, figureFileBig=MJsJeGiTFO2wYFEFoJB7qg==, tableContent=null), ArticleFig(id=1320890103845733102, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.5, caption=Effects of BC07 on cytokine levels in constipated mice (n=5), figureFileSmall=3kC9h40ymmR4xba92adwrA==, figureFileBig=mpN4hnHnu52cqvT7mqQJ1w==, tableContent=null), ArticleFig(id=1320890103921230575, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图5, caption=BC07对便秘小鼠细胞因子水平的影响(n=5), figureFileSmall=3kC9h40ymmR4xba92adwrA==, figureFileBig=mpN4hnHnu52cqvT7mqQJ1w==, tableContent=null), ArticleFig(id=1320890103984145136, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.6, caption=Treatment with BC07 alters the diversity and structure of the gut microbiota (n=5), figureFileSmall=O5iQDUazCvezV0cI6zbfWw==, figureFileBig=DpH+z/i0T7CqrkspbmcPLA==, tableContent=null), ArticleFig(id=1320890104055448305, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图6, caption=BC07干预对肠道微生物群的多样性和结构的影响(n=5)

注:A. ACE指数;B. Simpson指数;C. NMDS分析。

, figureFileSmall=O5iQDUazCvezV0cI6zbfWw==, figureFileBig=DpH+z/i0T7CqrkspbmcPLA==, tableContent=null), ArticleFig(id=1320890104122557170, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.7, caption=Effects of BC07 on the horizontal structure of intestinal microbiome in mice model of constipation (n=5), figureFileSmall=kLTAyxGqxr/7IKhK3R/+mQ==, figureFileBig=faO+5RRy52wxS/9QYt+eFw==, tableContent=null), ArticleFig(id=1320890104210637555, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图7, caption=BC07对便秘模型小鼠肠道微生物门水平结构的影响(n=5)

注:A. 门水平分布;B. 厚壁菌门占比;C. 拟杆菌门占比;D. 变形菌门占比;E. 厚壁菌门与拟杆菌门比值。

, figureFileSmall=kLTAyxGqxr/7IKhK3R/+mQ==, figureFileBig=faO+5RRy52wxS/9QYt+eFw==, tableContent=null), ArticleFig(id=1320890104315495156, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=EN, label=Fig.8, caption=Effects of BC07 on the Composition of gut microbiota at the genus level (n=5), figureFileSmall=zloIZvFUqGGxR8Tb1TuPRA==, figureFileBig=F1mFRvr+5WOAhgWKj0H5AA==, tableContent=null), ArticleFig(id=1320890104382604021, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320890091975852725, language=CN, label=图8, caption=BC07对肠道微生物属水平组成的影响(n=5)

注:A属水平分布;B不动杆菌属占比;C假单胞菌属占比;D毛螺菌属-NK4A136占比;E联合乳杆菌属占比。

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凝结海恩德里克斯氏菌BC07对盐酸洛哌丁胺诱导小鼠便秘的缓解作用
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孔素芬 , 朱明明 , 朱建国 , 方曙光 *
食品工业科技 | 营养与保健 2026,47(4): 435-442
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食品工业科技 |营养与保健 2026 , 47 (4) : 435 -442
凝结海恩德里克斯氏菌BC07对盐酸洛哌丁胺诱导小鼠便秘的缓解作用
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孔素芬 , 朱明明, 朱建国, 方曙光*
作者信息
  • 武汉微康益生菌研究院有限公司,湖北武汉 430000
通讯作者:
方曙光(1974−),男,博士,教授,研究方向:益生菌功能性、产业化关键技术开发,E-mail:。
作者简介:

孔素芬(1984−),女,本科,中级工程师,研究方向:益生菌菌种及功能开发研究,E-mail:

Relieving Effect of Heyndrickxia coagulans BC07 on Loperamide Hydrochloride-induced Constipation in Mice
Sufen KONG , Mingming ZHU, Jianguo ZHU, Shuguang FANG*
Affiliations
  • Wuhan Wecare Probiotic Research Institute Co., Ltd., Wuhan 430000, China
出版时间: 2026-02-15 doi: 10.13386/j.issn1002-0306.2025020073
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目的:研究凝结海恩德里克斯氏菌BC07对盐酸洛哌丁胺诱导小鼠便秘的影响。方法:将30只小鼠随机分为对照组(NC)、模型组(MC)、BC07组,每组10只,NC组和MC组灌胃生理盐水,BC07组灌胃凝结海恩德里克斯氏菌BC07菌液,持续14 d。第15~17 d使用盐酸洛哌丁胺造模,观察小鼠首粒黑便排出时间及肠转运情况,收集小鼠粪便,血清,盲肠和结肠样本。结果:摄入凝结海恩德里克斯氏菌BC07能够显著增加便秘小鼠的粪便水分含量(P<0.05),缩短小鼠首粒黑便排出时间(P<0.05),促进小肠的推进率;同时还能有效调节便秘小鼠胃动素(Motilin,MTL)、血管活性肠肽(Vasoactive intestinal peptide,VIP)、P物质(Substance P,SP)、生长抑素(Somatostatin,SS)的浓度,并能降低小鼠血清中细胞促炎因子IL-6和TNF-α的含量,提高抗炎因子IL-10的水平;此外,补充凝结海恩德里克斯氏菌BC07后,能有效调节便秘小鼠的肠道菌群结构,增加小鼠肠道有益菌群的相对丰度,显著降低有害菌群的相对丰度(P<0.05)。结论:凝结海恩德里克斯氏菌BC07具有提高粪便水分含量、减轻排便困难的作用,推测可能通过调节胃肠肽递质、改善肠道菌群结构、促进肠道蠕动来缓解便秘。

凝结海恩德里克斯氏菌BC07  /  便秘  /  调节

Objective: To investigate the effect of Heyndrickxia coagulans BC07 on loperamide hydrochloride-induced constipation in mice. Methods: Thirty mice were randomly divided into three groups: the normal control group (NC), the model group (MC) and the BC07 group, with 10 mice in each group. The NC and MC groups were intragastrically administered normal saline, while the BC07 group was intragastrically given Heyndrickxia coagulans BC07 bacterial solution for 14 days. From the 15th to the 17th day, loperamide hydrochloride was used to establish the constipation model. The time to the first black feces and intestinal transit in mice were observed. Feces, serum, cecum, and colon samples of mice were collected. Results: The intake of Heyndrickxia coagulans BC07 could significantly increase the feces water content in constipated mice (P<0.05), shorten the time to the first black feces in mice (P<0.05), and promote the propulsion rate of the small intestine. Meanwhile, it could effectively regulate the concentrations of motilin (MTL), vasoactive intestinal peptide (VIP), substance P (SP), and somatostatin (SS) in constipated mice. It also reduced the contents of pro-inflammatory cytokines IL-6 and TNF-α in the mouse serum and increased the level of the anti-inflammatory factor IL-10. In addition, after the supplementation of Heyndrickxia coagulans BC07, the structure of the intestinal microbiota in constipated mice could be effectively regulated, increasing the relative abundance of beneficial microbiota in the mouse intestine and significantly decreasing the relative abundance of harmful microflora (P<0.05). Conclusion: Heyndrickxia coagulans BC07 has the effect of increasing fecal water content and alleviating defecation difficulties. It is speculated that it may relieve constipation by regulating gastrointestinal peptide transmitters, improving the structure of the intestinal microbiota, and promoting intestinal peristalsis.

Heyndrickxia coagulans BC07  /  constipation  /  regulating
孔素芬, 朱明明, 朱建国, 方曙光. 凝结海恩德里克斯氏菌BC07对盐酸洛哌丁胺诱导小鼠便秘的缓解作用. 食品工业科技, 2026 , 47 (4) : 435 -442 . DOI: 10.13386/j.issn1002-0306.2025020073
Sufen KONG, Mingming ZHU, Jianguo ZHU, Shuguang FANG. Relieving Effect of Heyndrickxia coagulans BC07 on Loperamide Hydrochloride-induced Constipation in Mice[J]. Science and Technology of Food Industry, 2026 , 47 (4) : 435 -442 . DOI: 10.13386/j.issn1002-0306.2025020073
便秘是一种常见的功能性胃肠道疾病[1]。由于社会老龄化、饮食习惯改变以及生活压力的影响,便秘的发病率每年都在增长[2]。便秘会导致大量排泄物在肠道中积聚,以致毒素堆积,不仅影响生活质量,还可能导致其他健康问题,如痔疮、肠梗阻等[3]。目前,用于便秘的治疗方法主要包括药物和非药物治疗[4]。长时间服用溶剂性、刺激性泻药会带来严重副作用,导致肠道粘膜、肠神经和肠道菌群平衡遭到损害[5]。因此,开发缓解便秘的有效治疗方法对于改善全球范围内的健康状况至关重要。
凝结海恩德里克斯氏菌(也称凝结芽孢杆菌Bacillus coagulans)是一种产乳酸的芽孢杆菌,具有良好的益生特性,相比非芽孢杆菌的乳酸菌,凝结海恩德里克斯氏菌在胃肠道条件下具有较强的稳定性和耐受性[6]。凝结海恩德里克斯氏菌在生产、储存、运输和应用方面具有高度的稳定性,使其成为食品和药品配方中更合适的候选者[7]。凝结海恩德里克斯氏菌在食品中的安全使用历史悠久,并且已获得美国食品药品监督管理局(FDA)授予的“普遍认为安全”(GRAS)状态[8]。Jung等[9]研究表明,凝结海恩德里克斯氏菌SNZ1969的摄入可通过粘蛋白的分泌和胃肠道激素的改变减轻便秘。同样,刘文英等[10]发现,凝结海恩德里克斯氏菌B.C-39复合微生态制剂可以使胃肠激素的分泌趋于正常,并增加小鼠肠道SCFAs(Short-Chain Fatty Acids)的含量,具有缓解便秘的作用。
凝结海恩德里克斯氏菌BC07分离于中国四川省的泡菜,保藏于中国典型培养物保藏中心,编号CCTCCNO:M2022195。基于凝结海恩德里克斯氏菌BC07前期的研究[11],显示其具有较强的抗逆性和稳定性,但是还未开展改善便秘,调节肠道菌群的体内研究。本研究考察了凝结海恩德里克斯氏菌BC07对缓解盐酸洛哌丁胺诱导的功能性便秘的作用,并初步讨论了其影响机制,为后续益生菌在保健食品和药物中的应用提供了实验基础和理论依据。
盐酸洛哌丁胺胶囊 西安杨森制药有限公司;活性炭粉 国药集团化学制药有限公司;阿拉伯胶 上海麦克林生化科技有限公司;福尔马林组织固定液 武汉赛维尔生物科技有限公司;酶联免疫ELISA试剂盒 武汉华联科生物技术有限公司;PCA固体培养基 青岛海博生物技术有限公司;生理盐水 四川科伦药业股份有限公司;凝结海恩德里克斯氏BC07菌株 保藏于中国典型培养物保藏中心;小鼠 购自斯贝福(北京)生物技术有限公司,许可证号:SCXK(京)2019-0010。
CX-23显微镜 奥林巴斯(广州)工业有限公司;UV-1700紫外分光光度计 上海美析仪器有限公司;SPX-250B-2型恒温培养箱 上海福玛实验设备有限公司;EF28 pH计 梅特勒-托利多(上海)有限公司;JY1002电子天平 上海衡平仪器仪表厂; DP812 Qubit dsDNA HS检测试剂盒 天根生化科技(北京)有限公司。
制备凝结海恩德里克斯氏菌BC07菌悬液:将活化后的BC07涂布接种于PCA固体培养基中,42 ℃培养72 h,洗脱收集菌体制成109 CFU/mL的菌悬液,置于−80 ℃冰箱保存备用[11]。
将阿拉伯胶100 g与水800 mL混合,加热煮沸至透明溶液,加入活性炭50 g,煮沸三次,待溶液凉后加水定容至1000 mL,于4 ℃冰箱中保存,用前摇匀[12]。
6~8周龄SPF级BALB/c雄性小鼠(20~22 g),在室温为25±2 ℃,湿度为50%±5%,光-暗循环12 h环境中饲养[13]。实验动物福利伦理审查编号:安评中心动(福)第202310248号。试验期间各组小鼠均饲喂标准饲料,经过7 d的适应期后,将30只小鼠随机分为对照组(NC)、模型组(MC)、BC07组,每组10只,从适应期结束后的第1 d开始干预,NC组和MC组每只小鼠灌胃生理盐水200 µL,BC07组每只小鼠灌胃凝结海恩德里克斯氏菌BC07菌液200 µL(5×109 CFU/mL),持续14 d。第15~17 d使用盐酸洛哌丁胺造模,MC组、BC07组按小鼠体重10 mg/kg.bw灌胃盐酸洛哌丁胺,NC组灌胃等体积的生理盐水。4 h后,NC组、MC组灌胃生理盐水,BC07组灌胃BC07菌液。第18 d观察小鼠首粒黑便排出时间,收集小鼠粪便,第19 d采集眼球血后处死小鼠,观察肠转运情况,并收集盲肠、结肠样本。
第17 d每只小鼠都被转移到一个干净的空笼子里,收集3 h粪便样本,收集期间记录每只小鼠的粪便粒数,并将粪便装入2 mL离心管中称量粪便湿重和干重,测定水分含量[3]。
$ \mathrm{粪便含水量}(\text{%})=\frac{\text{粪便湿重}-\text{粪便干重}}{\text{粪便湿重}}\times 100 $
第18 d(禁食18 h后),NC组小鼠灌胃生理盐水,其他组灌胃盐酸洛哌丁胺,1 h后NC组和MC组灌胃墨汁,BC07组灌胃含凝结海恩德里克斯氏菌BC07菌液的墨汁。将每只小鼠分别转移到干净的空笼子里,并提供饲料和水,从灌胃墨汁开始计时,记录每只小鼠首粒黑便排出时间,以模型组最后一只小鼠的首粒黑便排出时间为终止时间[14]。
在小鼠灌胃的第19 d(禁食18 h后),NC组小鼠灌胃生理盐水,其他组的小鼠灌胃盐酸洛哌丁胺[14]。1 h后,NC组、MC组灌胃墨汁,BC07组灌胃含凝结海恩德里克斯氏菌BC07的墨汁。20 min后处死小鼠,打开腹腔,分离肠系膜,剪取上端至幽门,下端至回盲肠的肠管,轻轻将小肠拉成直线,测量肠管长度为小肠总长度,从幽门到墨汁前沿为墨汁推进长度。小肠推进率计算公式如下:
$ \mathrm{小肠推进率}(\text{%})=\frac{\mathrm{墨汁推进长度}(\text{cm})}{\mathrm{小肠总长度}(\mathrm{cm})}\times 100 $
取出小鼠的结肠组织,剪取1 cm结肠置于无菌离心管中,用10%甲醛溶液固定过夜后进行H&E染色,使用光学显微镜观察染色后的结肠组织[15]。
将收集到的小鼠血液静置2 h,845×g离心10 min吸取血清,采用ELISA试剂盒检测小鼠血清中与便秘相关的胃肠道调节递质水平:小鼠胃动素(MTL)、血管活性肠肽(VIP)、P物质(SP)、生长抑素(SS);炎症因子白细胞介素IL-10、IL-6、TNF-α[16]。
采用Qubit dsDNA HS检测试剂盒的方法,提取各组小鼠盲肠粪便中微生物基因组DNA,使用Qubit dsDNA HS检测试剂盒和Qubit 4.0荧光仪检测样本DNA浓度[17]。对样本中所有细菌的V3~V4高变区在Illumina NovaSeq 6000测序平台上进行16S rDNA测序,使用百迈客云平台对生物信息学进行分析,确定肠道微生物群的特征。
每组重复三次试验,结果以平均值±标准差表示;使用SPSS 23.0软件进行显著性分析,组间比较采用单因素方差分析(One-Way ANOVA),事后比较采用Tukey检验;P<0.05被认为有统计学意义,数据和制图采用GraphPad Prism V8.4.3.686软件进行处理。
排便困难,频率低,粪便坚硬干结是便秘的常见症状[17]。本试验通过测定盐酸洛哌丁胺造模3 d后的小鼠粪便颗粒数和含水量,观察不同组别小鼠便秘的程度。如图1A所示,与NC组粪便颗粒数相比,MC组粪便颗粒数显著减少(P<0.05),表明盐酸洛哌丁胺处理降低了小鼠排便的颗粒数,造成了小鼠便秘的结果。经BC07干预后小鼠粪便颗粒数与MC组粪便颗粒数无显著差异,此结果与Jang等[18]的研究中低剂量组结果相近,而当提高灌胃菌液浓度后,小鼠排便数量显著高于MC组。
各组小鼠粪便含水量如图1B所示,MC组的粪便含水量低于NC组,具有统计学差异(P<0.05),说明盐酸洛哌丁胺给药后可导致结肠对水分的过度吸收,降低结肠内容物含水率,导致便秘[19]。相比MC组,凝结海恩德里克斯氏菌BC07干预后,粪便水分含量显著增加(P<0.05),表明凝结海恩德里克斯氏菌BC07能增加便秘小鼠的粪便水分含量,从而软化粪便,减轻排便困难。
小鼠首次排出黑便时间和小肠推进率反映了整个肠道的运动性。也就是说,首次排出黑便的时间越短,小肠推进率越高,表明整个肠道的运动速度越快,肠道运输能力越强[11]。如图2A所示,盐酸洛哌丁胺造模4 d后,NC组首粒黑便排出时间59 min,MC组黑便时间达到了174 min,显著长于NC组(P<0.05),说明灌胃盐酸洛哌丁胺导致了整个肠道的蠕动能力和运输能力下降,从而延长了排便时间。而经凝结海恩德里克斯氏菌BC07干预后,小鼠首粒黑便时间缩短,且显著低于MC组(P<0.05),该结果与刘文英等[10]研究一致。表明凝结海恩德里克斯氏菌BC07可以通过改善肠道功能和肠道运动提高便秘小鼠的肠道传输率,从而缩短首粒黑便排出时间。
肠道推进率是指单位时间内消化物在肠道内的运动速率,是研究小鼠便秘情况的重要指标之一[20]。小肠运动主要是通过蠕动力推动肠内容物运动。较高的胃肠道推进率表明食糜在小肠中的停留时间较短,且小肠的运动性较强。实验结果如图2B所示,盐酸洛哌丁胺处理后,MC组的胃肠道推进率低于NC组(P<0.05),表明便秘模型建立成功。灌胃凝结海恩德里克斯氏菌BC07后,便秘小鼠小肠推进率显著提升(P<0.05)。Zhou等[21]在便秘昆明鼠的研究中体现了同样的结果,表明凝结海恩德里克斯氏菌BC07可以有效改善肠道蠕动,并能加速食糜通过小肠的运动和排出。综合以上结果,表明凝结海恩德里克斯氏菌BC07在推进小肠运动,促进粪便排出方面具有积极作用。
观察小鼠结肠组织切片如图3所示,NC组结肠黏膜的上皮完整,细胞排列整齐,未见萎缩、糜烂、坏死,小鼠结肠形态健康。MC组经盐酸洛哌丁胺处理后,结肠组织大量炎细胞浸润,结肠黏膜变薄,杯状细胞显著减少。而经凝结海恩德里克斯氏菌BC07干预后,小鼠的结肠组织得到恢复,炎细胞浸润减少,细胞排列趋向整齐。便秘和肠易激综合征(IBS)常常由于上皮和粘膜层的变化,而破坏产生表达粘蛋白和糖蛋白的胃肠道屏障,补充益生菌可以增加小肠隐窝高度、肠干细胞、潘氏细胞和杯状细胞的数量[17]。
神经递质是神经细胞分泌的化学物质,是调节肠道运动的重要因素之一,肠神经递质分泌失衡可引起肠功能失调而导致疾病发生[22]。研究便秘小鼠血清中胃肠神经递质如图4所示,结果发现,与NC组相比,MC组小鼠VIP、SS含量显著升高(P<0.05),MTL和SP含量显著降低(P<0.05),表明盐酸洛哌丁胺诱导小鼠便秘与血清胃肠神经递质浓度变化相关。Zhou等[21]发现BC01可以通过调节兴奋性胃肠激素和抑制性胃肠激素的分泌水平来加速胃肠道运输并缓解便秘。本研究通过灌胃凝结海恩德里克斯氏菌BC07对便秘小鼠进行干预,结果显示,相比MC组,BC07组小鼠VIP、SS含量显著降低(P<0.05),MTL和SP含量显著升高(P<0.05),使胃肠神经递质浓度趋于正常水平,缓解小鼠便秘发生。
通过测定便秘小鼠血清中细胞炎症因子TNF-α、IL-6、IL-10的水平,定量研究BC07对便秘小鼠血清中炎症因子水平的影响。结果如图5所示,MC组便秘小鼠的抗炎因子(IL-10)含量显著低于NC组(P<0.05),促炎因子IL-6浓度高于NC组,促炎因子TNF-α水平显著高于NC组(P<0.05),表明便秘造成了小鼠血清细胞炎症因子的紊乱,而BC07组抗炎因子IL-10的水平显著增加(P<0.05),促炎因子IL-6、TNF-α的含量降低。Jang等[23]研究发现发酵粘液乳杆菌KBL375显著降低促炎因子IL-1β、TNF-α和IL-6并升高抗炎细胞因子IL-10减少肠道炎症。在本研究中,通过凝结海恩德里克斯氏菌BC07干预,下调了IL-6和TNF-α的表达,显著提升了抗炎因子IL-10的含量(P<0.05),抑制了炎症反应,结果表明,BC07可能通过预防肠道损伤缓解便秘。
对各组肠道菌群α多样性进行分析如图6A和图6B所示,小鼠肠道微生物α多样性中MC组ACE指数,Simpson指数相比NC组均降低。相比MC组,BC07组的肠道微生物α多样性中的ACE指数、Simpson指数均升高,且后者具有显著性差异(P<0.05),说明盐酸洛哌丁胺诱导的小鼠便秘造成了小鼠肠道菌群物种丰富度和多样性的下降,而凝结海恩德里克斯氏菌BC07的干预可提高肠道微生物菌群α多样性。同样,便秘也导致了MC组肠道微生物群β多样性的变化,如图6C所示,MC组菌群结构和NC组菌群结构出现差异,经凝结海恩德里克斯氏菌BC07干预后,便秘小鼠肠道微生物群落结构趋于正常对照组。先前的研究发现,哺乳动物肠道微生物组成与益生菌摄入相关联[24]。本研究的结果显示,BC07的干预对便秘小鼠肠道微生物群多样性和结构产生影响,微生物群组成的差异可能影响便秘症状。
通过对小鼠肠道微生物门水平的研究发现,如图7所示,盐酸洛哌丁胺处理后MC组的厚壁菌门相对丰度显著降低(P<0.05),拟杆菌门丰度无显著变化,厚壁菌门与拟杆菌门的比值降低(P<0.05)。已知厚壁菌门/拟杆菌门(F/B)的比例对维持肠道内稳态有重要影响[25]。当F/B比率失衡时,各种疾病如炎症、自身免疫性疾病和癌症可能发生[26]。此外,MC组变形菌门相对丰度显著升高(P<0.05),变形菌门是肠道细菌中最大的门之一,包括很多病原菌,如大肠杆菌、沙门氏菌、霍乱弧菌、幽门螺杆菌等。变形菌门的比例增加是微生物群失调的标志,也是疾病的潜在诊断标准[27]。本研究结果显示,便秘造成了小鼠肠道有害菌群的增加,破坏了小鼠肠道菌群平衡,凝结海恩德里克斯氏菌BC07干预后,厚壁菌门相对丰度显著升高(P<0.05),变形菌门相对丰度显著降低(P<0.05),表明凝结海恩德里克斯氏菌BC07具有调节小鼠肠道菌群结构变化,缓解小鼠便秘的作用。
经OTU聚类分析发现,属水平肠道微生物存在显著差异,如图8所示,和NC组相比,MC组毛螺菌属_NK4A136相对丰度降低,而有害菌属不动杆菌属、假单胞菌属相对丰度升高(P<0.05)。凝结海恩德里克斯氏菌BC07干预后有益菌属毛螺菌属_NK4A136、联合乳杆菌属相对丰度增加,有害菌属不动杆菌属、假单胞菌属相对丰度显著下降(P<0.05),这与Wang等[1]研究结果一致。毛螺菌属_NK4A136(Lachnospiraceae_NK4A136)属于厌氧、产孢细菌的毛螺菌科,Lachnospiraceae_NK4A136可通过发酵膳食多糖产生SCFAs,与多种代谢性疾病和慢性炎症呈负相关[28];联合乳杆菌属(Ligilactobacillus)被认为对人类健康有益,可能有助于改善肠道健康、增强免疫功能和抑制有害细菌的生长[29];不动杆菌属(Acinetobacter),革兰氏阴性菌,可定植于人体皮肤、伤口、呼吸道和胃肠道,也可存在于口腔生物膜,具有高致病性并导致侵袭性疾病[30]。假单胞菌属(Pseudomonas)与肠道屏障功能障碍和感染有关,可能导致败血症和多器官功能障碍综合征。此外,假单胞菌与炎症性肠病的发生也有关系,在克罗恩病患者体内频繁检测到假单胞菌[31]。
本研究以盐酸洛哌丁胺诱导小鼠便秘为模型,通过灌胃凝结海恩德里克斯氏菌BC07观察对小鼠便秘的改善效果。结果表明,凝结海恩德里克斯氏菌BC07能够显著增加便秘小鼠的粪便水分含量,缩短小鼠首粒黑便排出时间,提高食糜在小肠的推进率。此外,凝结海恩德里克斯氏菌BC07干预后,小鼠结肠组织得到恢复。通过血清生化分析,灌胃BC07能有效调节便秘小鼠的胃肠肽递质VIP、SS、MTL、SP的浓度,并能降低小鼠血清中细胞促炎因子IL-6、TNF-α的含量,提高抗炎因子IL-10的水平。进一步的,补充凝结海恩德里克斯氏菌BC07后,小鼠肠道内有益菌群相对丰度增加,有害菌群相对丰度显著降低,BC07有效改善了便秘小鼠的肠道菌群结构。初步推测,凝结海恩德里克斯氏菌BC07通过调节神经肽递质的水平,增强肠道的蠕动,间接影响肠道健康,改善消化功能,促进排便。本研究提供了凝结海恩德里克斯氏菌BC07缓解便秘的基础数据,但是由于机体的复杂性,需要综合分析改善便秘的内在机制如肠道菌群与神经肽递质的相互关系,通过菌-肠-脑轴研究特定菌群代谢产物如短链脂肪酸、脂多糖等的变化影响神经肌肉功能,进一步挖掘凝结海恩德里克斯氏菌BC07改善便秘的作用机制。

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doi: 10.13386/j.issn1002-0306.2025020073
  • 接收时间:2025-02-12
  • 首发时间:2026-10-24
  • 出版时间:2026-02-15
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  • 收稿日期:2025-02-12
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    武汉微康益生菌研究院有限公司,湖北武汉 430000

通讯作者:

方曙光(1974−),男,博士,教授,研究方向:益生菌功能性、产业化关键技术开发,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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