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Article(id=1297570993610969957, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260055, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1768838400000, receivedDateStr=2026-01-20, revisedDate=null, revisedDateStr=null, acceptedDate=1772121600000, acceptedDateStr=2026-02-27, onlineDate=1787294632579, onlineDateStr=2026-08-21, pubDate=1785772800000, pubDateStr=2026-08-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1787294632579, onlineIssueDateStr=2026-08-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1787294632579, creator=13701087609, updateTime=1787294632579, updator=13701087609, issue=Issue{id=1297570992835023717, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='8', pageStart='3681', pageEnd='4288', issueExtLink='null', onlineDate='null', pubDate='1785772800000', pubDateStr='2026-08-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1787294632395, creator='13701087609', updateTime=1787294931551, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1297572247670124783, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1297572247670124784, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=3913, endPage=3925, ext={EN=ArticleExt(id=1297570993816490854, articleId=1297570993610969957, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Protective effects of Camphora kanahirae leaves on gut microbiota and short-chain fatty acid metabolism in the mouse model of alcoholic liver disease, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] To investigate the mechanism by which Camphora kanahirae leaves alleviate alcoholic liver disease through the regulation of the gut microbiota-short-chain fatty acid-intestinal barrier pathway. [Methods] Sixty mice were randomly allocated into six groups [normal, model, silymarin (100 mg/kg), and low-, medium-, and high-dose (100, 200, and 400 mg/kg, respectively) C. kanahirae leaves], with 10 mice in each group. After 14 consecutive days of administration, a mouse model of alcoholic liver disease was established. The liver index, alanine aminotransferase (ALT) and aspartate transferase (AST) levels, colon histopathological changes, short-chain fatty acid content, and alterations in gut microbiota structure were measured. [Results] Compared with the model group, medium- and high-dose C. kanahirae leaves reduced ALT and AST levels (P<0.05). Pathological evaluation showed that C. kanahirae leaves at all doses alleviated alcohol-induced colon mucosal damage and inflammatory cell infiltration to varying degrees. Alcohol damage resulted in decreases in levels of short-chain fatty acids, such as acetic acid, propionic acid, butyric acid, and valeric acid (P<0.05). C. kanahirae leaves promoted the proliferation of beneficial bacteria, increased short-chain fatty acid levels, and enhanced intestinal barrier function. Furthermore, 16S rRNA gene analysis of the gut microbiota indicated that alcohol intake led to dysbiosis, and C. kanahirae leaves effectively reversed this microbial imbalance, restoring the composition and diversity of the gut microbiota. [Conclusion] C. kanahirae leaves exert hepatoprotective effects by regulating the gut microbiota balance, promoting short-chain fatty acid production, and maintaining intestinal barrier integrity.

, authors=Peiyao TAN1, Xuezheng LIANG2, Jianwei LÜ3, Xiaomin YANG4, Bei ZHANG3, authorsList=Peiyao TAN, Xuezheng LIANG, Jianwei LÜ, Xiaomin YANG, Bei ZHANG, authorCompany=null, correspAuthors=Bei ZHANG, authorNote=null, correspAuthorsNote=
E-mail:
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【目的】 探讨牛樟叶通过调节“肠道菌群-短链脂肪酸-肠道屏障”途径缓解酒精性肝病的作用机制。 【方法】 将60只小鼠随机分为6组[正常组、模型组、水飞蓟宾(100 mg/kg)组、牛樟叶低(100 mg/kg)组、牛樟叶中(200 mg/kg)组、牛樟叶高(400 mg/kg)组]，每组10只，连续给药14 d后建立酒精性肝病小鼠模型，检测肝脏指数、丙氨酸氨基转移酶(alanine aminotransferase, ALT)和天冬氨酸氨基转移酶(aspartate aminotransferase, AST)水平、结肠组织病理变化、短链脂肪酸含量及肠道菌群结构变化。 【结果】 与模型组相比，牛樟叶中、高剂量组显著降低ALT和AST水平(P<0.05)；病理学评估显示，各剂量牛樟叶均能不同程度减轻酒精导致结肠黏膜损伤及炎性细胞浸润；酒精损伤导致乙酸、丙酸、丁酸和戊酸等短链脂肪酸含量显著下降(P<0.05)，牛樟叶可促进有益菌增殖，提高短链脂肪酸水平，增强肠道屏障功能；16S rRNA基因测序分析显示，酒精摄入导致肠道菌群失调，牛樟叶能有效逆转菌群紊乱，恢复菌群组成与多样性。 【结论】 牛樟叶通过调节肠道菌群平衡、促进短链脂肪酸生成及维护肠道屏障完整性发挥肝保护作用。

, authors=谭培姚1, 梁学政2, 吕建伟3, 杨晓敏4, 张蓓3, authorsList=谭培姚, 梁学政, 吕建伟, 杨晓敏, 张蓓, authorCompany=null, correspAuthors=张蓓, authorNote=

作者贡献声明

谭培姚：数据收集及统计分析，撰写文章；梁学政：提供资源，监督管理；吕建伟：提出概念，方法论；杨晓敏：规范分析，方法设计；张蓓：实验设计与指导，项目管理及获取基金。

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Contents of short-chain fatty acids in cecal contents/(μg/g)

, figureFileSmall=null, figureFileBig=null, tableContent=
GroupAcetic acidPropionic acidButyric acidValeric acidCaproic acidIsobutyric acidIsovaleric acid
NC1 709.77±6.281 062.61±10.621 911.09±15.84213.42±3.6518.57±0.19149.16±1.32169.18±3.01
ALD922.01±16.30#617.72±7.35#656.25±7.60#126.87±0.78#9.42±0.14#112.75±4.12#113.14±3.87#
Sil1 584.67±10.31*734.98±8.03*1 464.41±12.65*179.83±6.32*18.63±1.30*134.38±5.35*139.52±3.70*
CKH1 145.48±8.10*897.21±10.37*920.15±10.35*194.22±4.15*19.93±1.54*129.32±2.10*157.01±2.10*
), ArticleFig(id=1297571002733581242, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297570993610969957, language=CN, label=表1, caption=

盲肠内容物中短链脂肪酸含量

, figureFileSmall=null, figureFileBig=null, tableContent=
GroupAcetic acidPropionic acidButyric acidValeric acidCaproic acidIsobutyric acidIsovaleric acid
NC1 709.77±6.281 062.61±10.621 911.09±15.84213.42±3.6518.57±0.19149.16±1.32169.18±3.01
ALD922.01±16.30#617.72±7.35#656.25±7.60#126.87±0.78#9.42±0.14#112.75±4.12#113.14±3.87#
Sil1 584.67±10.31*734.98±8.03*1 464.41±12.65*179.83±6.32*18.63±1.30*134.38±5.35*139.52±3.70*
CKH1 145.48±8.10*897.21±10.37*920.15±10.35*194.22±4.15*19.93±1.54*129.32±2.10*157.01±2.10*
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牛樟叶对酒精性肝病小鼠肠道菌群和短链脂肪酸代谢的保护作用
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谭培姚 1 , 梁学政 2 , 吕建伟 3 , 杨晓敏 4 , 张蓓 3
微生物学报 | 研究报告 2026,66(8): 3913-3925
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微生物学报 |研究报告 2026 , 66 (8) : 3913 -3925
牛樟叶对酒精性肝病小鼠肠道菌群和短链脂肪酸代谢的保护作用
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谭培姚1, 梁学政2, 吕建伟3, 杨晓敏4, 张蓓3
作者信息
  • 1.广西中医药大学，广西 南宁
  • 2.柳州市中医医院(柳州市壮医医院)，广西 柳州
  • 3.广西中医药大学第一附属医院，广西 南宁
  • 4.柳州市妇幼保健院，广西 柳州
通讯作者:
张蓓
作者简介:

作者贡献声明

谭培姚：数据收集及统计分析，撰写文章；梁学政：提供资源，监督管理；吕建伟：提出概念，方法论；杨晓敏：规范分析，方法设计；张蓓：实验设计与指导，项目管理及获取基金。

Protective effects of Camphora kanahirae leaves on gut microbiota and short-chain fatty acid metabolism in the mouse model of alcoholic liver disease
Peiyao TAN1, Xuezheng LIANG2, Jianwei LÜ3, Xiaomin YANG4, Bei ZHANG3
Affiliations
  • 1.Guangxi University of Chinese Medicine, Nanning, Guangxi, China
  • 2.Liuzhou Traditional Chinese Medicine Hospital, Liuzhou, Guangxi, China
  • 3.The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, Guangxi, China
  • 4.Liuzhou Maternity and Child Healthcare Hospital, Liuzhou, Guangxi, China
    • Corresponding Author:
    E-mail:
出版时间: 2026-08-04 doi: 10.13343/j.cnki.wsxb.20260055
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【目的】 探讨牛樟叶通过调节“肠道菌群-短链脂肪酸-肠道屏障”途径缓解酒精性肝病的作用机制。 【方法】 将60只小鼠随机分为6组[正常组、模型组、水飞蓟宾(100 mg/kg)组、牛樟叶低(100 mg/kg)组、牛樟叶中(200 mg/kg)组、牛樟叶高(400 mg/kg)组]，每组10只，连续给药14 d后建立酒精性肝病小鼠模型，检测肝脏指数、丙氨酸氨基转移酶(alanine aminotransferase, ALT)和天冬氨酸氨基转移酶(aspartate aminotransferase, AST)水平、结肠组织病理变化、短链脂肪酸含量及肠道菌群结构变化。 【结果】 与模型组相比，牛樟叶中、高剂量组显著降低ALT和AST水平(P<0.05)；病理学评估显示，各剂量牛樟叶均能不同程度减轻酒精导致结肠黏膜损伤及炎性细胞浸润；酒精损伤导致乙酸、丙酸、丁酸和戊酸等短链脂肪酸含量显著下降(P<0.05)，牛樟叶可促进有益菌增殖，提高短链脂肪酸水平，增强肠道屏障功能；16S rRNA基因测序分析显示，酒精摄入导致肠道菌群失调，牛樟叶能有效逆转菌群紊乱，恢复菌群组成与多样性。 【结论】 牛樟叶通过调节肠道菌群平衡、促进短链脂肪酸生成及维护肠道屏障完整性发挥肝保护作用。

牛樟叶  /  酒精性肝病  /  短链脂肪酸  /  肠道菌群

[Objective] To investigate the mechanism by which Camphora kanahirae leaves alleviate alcoholic liver disease through the regulation of the gut microbiota-short-chain fatty acid-intestinal barrier pathway. [Methods] Sixty mice were randomly allocated into six groups [normal, model, silymarin (100 mg/kg), and low-, medium-, and high-dose (100, 200, and 400 mg/kg, respectively) C. kanahirae leaves], with 10 mice in each group. After 14 consecutive days of administration, a mouse model of alcoholic liver disease was established. The liver index, alanine aminotransferase (ALT) and aspartate transferase (AST) levels, colon histopathological changes, short-chain fatty acid content, and alterations in gut microbiota structure were measured. [Results] Compared with the model group, medium- and high-dose C. kanahirae leaves reduced ALT and AST levels (P<0.05). Pathological evaluation showed that C. kanahirae leaves at all doses alleviated alcohol-induced colon mucosal damage and inflammatory cell infiltration to varying degrees. Alcohol damage resulted in decreases in levels of short-chain fatty acids, such as acetic acid, propionic acid, butyric acid, and valeric acid (P<0.05). C. kanahirae leaves promoted the proliferation of beneficial bacteria, increased short-chain fatty acid levels, and enhanced intestinal barrier function. Furthermore, 16S rRNA gene analysis of the gut microbiota indicated that alcohol intake led to dysbiosis, and C. kanahirae leaves effectively reversed this microbial imbalance, restoring the composition and diversity of the gut microbiota. [Conclusion] C. kanahirae leaves exert hepatoprotective effects by regulating the gut microbiota balance, promoting short-chain fatty acid production, and maintaining intestinal barrier integrity.

Camphora kanahirae leaf  /  alcoholic liver disease  /  short-chain fatty acid  /  gut microbiota
谭培姚, 梁学政, 吕建伟, 杨晓敏, 张蓓. 牛樟叶对酒精性肝病小鼠肠道菌群和短链脂肪酸代谢的保护作用. 微生物学报, 2026 , 66 (8) : 3913 -3925 . DOI: 10.13343/j.cnki.wsxb.20260055
Peiyao TAN, Xuezheng LIANG, Jianwei LÜ, Xiaomin YANG, Bei ZHANG. Protective effects of Camphora kanahirae leaves on gut microbiota and short-chain fatty acid metabolism in the mouse model of alcoholic liver disease[J]. Acta Microbiologica Sinica, 2026 , 66 (8) : 3913 -3925 . DOI: 10.13343/j.cnki.wsxb.20260055
正文
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酒精性肝病(alcoholic liver disease, ALD)是全球范围内不容忽视的公共健康问题，其持续加剧的疾病负担与过早死亡率已引起广泛关注[1]。酒精性肝病始于肝脏的早期损伤，若未及时干预，可逐步进展为脂肪性肝炎、肝纤维化乃至肝细胞癌等严重肝脏病变[2]。然而，酒精性肝病的早期诊断面临挑战，致使多数病例在进展至晚期才被确诊，加之目前缺乏有效的治疗药物，患者面临较高的疾病恶化风险[3]。因此，深入研究酒精性肝病所致短链脂肪酸(short-chain fatty acid, SCFA)代谢异常及其肠道菌群结构紊乱，并利用16S rRNA基因测序技术阐明其作用机制，对开发针对酒精性肝病的早期干预药物具有重要的临床意义。
近年来，“肠-肝轴”理论的提出为深入理解酒精性肝病的发病机制提供了新的视角。作为连接肝脏与肠道的关键桥梁，肠道微生物组是一个由细菌、真菌、病毒及古细菌共同构成的复杂生态系统[4]，这些微生物栖息于胃肠道内，通过门静脉循环与肝脏进行双向交流。肠道微生物可将膳食纤维经厌氧发酵生成短链脂肪酸[5]。短链脂肪酸不仅能为肠道菌群与宿主肠上皮细胞提供能量来源，还能调节肠道免疫、抑制病原体生长并减轻炎症反应[6]。研究表明，急性酒精暴露会直接破坏上皮紧密连接的完整性，诱发肠道菌群失调，并可能引发氧化还原比失衡，减少了短链脂肪酸的生成，干扰其在肠上皮细胞中的代谢过程，最终导致肠道黏膜屏障功能受损[7-8]。
肠道屏障功能一旦受损，内毒素等代谢产物即可易位至肝脏，激活肝脏免疫细胞，引发炎症反应从而加剧肝损伤[9]。此外，菌群失调所致短链脂肪酸代谢紊乱通过破坏肠屏障、激活炎症反应和干扰表观遗传调控等多种途径，在肝脏疾病的发生和发展中起着关键作用[10]。综上所述，深入分析“肠道菌群-短链脂肪酸-肝脏”这一分子机制可为酒精性肝病的防治提供新的思路。
牛樟(Camphora kanahirae)作为中国台湾地区特有的植物，其叶片中富含的多糖成分具有多种生物活性[11]。然而，关于牛樟叶是否通过调控“肠道菌群-短链脂肪酸-肠道屏障”改善酒精性肝病，目前尚缺乏相关研究。为此，本研究旨在通过检测肝脏指数、丙氨酸氨基转移酶和天冬氨酸氨基转移酶水平、结肠组织病理变化、肠道菌群结构及短链脂肪酸含量探讨牛樟叶对酒精性肝病的保护作用及机制，分析其对肠道菌群组成、短链脂肪酸生成及肠道屏障功能的影响，并通过检测相关生化指标及观察肝脏组织病理学变化，揭示其经由肠-肝轴发挥作用的机制，为牛樟叶的进一步开发与应用提供理论依据。
1 材料与方法
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1.1 动物
60只雄性SPF级昆明小鼠(KM小鼠，8周龄)购自浙江维通利华实验动物技术有限公司桐乡分公司，合格证号SCXK(浙)2020-0002。本研究所有动物实验获得广西中医药大学动物伦理委员会批准，编号为KS20250000-266。
1.2 主要试剂和仪器
牛樟叶，柳州市天姿园艺有限公司，经广西中医药大学药学院杨柯教授鉴定为牛樟的叶子；水飞蓟宾，上海阿拉丁生化科技股份有限公司；丙氨酸氨基转移酶和天冬氨酸氨基转移酶试剂盒，南京建成生物工程研究所有限公司；磷酸、甲基叔丁基醚(methyl tertiary butyl ether, MTBE)，上海安谱实验科技股份有限公司；无水乙醇，国药集团化学试剂有限公司；VAHTSTM DNA Clean Beads试剂，南京诺唯赞生物科技股份有限公司；Quant-iT PicoGreen dsDNA检测试剂盒，赛默飞世尔科技公司；Soil DNA Kit，Omega Bio-Tek公司。
高速多功能粉碎机，武义海纳电器有限公司；电热恒温水浴锅，上海精宏实验设备有限公司；旋转蒸发仪，西安中凯科教仪器有限公司；全波长酶标仪，赛默飞世尔科技公司；正置显微镜、石蜡切片机，徕卡显微系统(上海)有限公司；高速冷冻离心机，杭州奥盛仪器有限公司；摊片、烤片机，湖北康强医疗器械有限公司；生物组织包埋机-冷冻机，湖北泰维科技实业股份有限公司；气相色谱-质谱仪(gas chromatography-mass spectrometry, GC-MS)，安捷伦科技(中国)有限公司。
1.3 牛樟叶提取物的制备
称取5 kg干燥牛樟叶，粉碎后过60目筛，按1 g:40 mL料液比加入蒸馏水，90 ℃条件下回流提取3次，每次75 min，合并滤液，加热浓缩，加入75%乙醇沉淀36 h，抽滤得牛樟叶提取物，备用。为明确提取物化学特征，参照前期工艺[12]，采用苯酚-硫酸法测定多糖含量，结果为8.6 mg/g。
1.4 酒精性肝病小鼠模型的建立
KM小鼠适应性饲养7 d后，随机分为6组：正常组(NC组)、模型组(ALD组)、水飞蓟宾组(Sil组，100 mg/kg)、牛樟叶低剂量组(CKL组，100 mg/kg)、牛樟叶中剂量组(CKZ组，200 mg/kg)和牛樟叶高剂量组(CKH组，400 mg/kg)，每组10只。除正常组与模型组灌胃等量生理盐水外，其余各组灌胃等体积相应药物，连续14 d。灌胃结束后，除正常组继续灌胃生理盐水外，其余各组灌胃20 mL/kg的56°白酒。
1.5 动物取材
干预结束后，采用小鼠应激排便法收集粪便，每组各取3个样本，置于无菌冻存管内，于-80 ℃冰箱保存备用。禁食禁水12 h后，麻醉取血并取肝脏组织备用；另取3 cm空肠和结肠组织置于4%多聚甲醛中固定，同时取出肠道内容物置于-80 ℃保存。
1.6 肝脏指数
准确称量各组小鼠肝脏质量，按公式(1)计算肝指数。
肝指数=小鼠肝脏质量/小鼠体质量×100%
1.7 生化指标检测
采用酶联免疫吸附试验(enzyme-linked immunosorbent assay, ELISA)法，使用相应试剂盒检测小鼠血清丙氨酸氨基转移酶和天冬氨酸氨基转移酶水平。
1.8 结肠组织病理学观察
将结肠组织固定于多聚甲醛中后，按照常规病理程序进行苏木精-伊红染色(hematoxylin and eosin, HE)，于光学显微镜下观察病理形态。
1.9 盲肠内容物中短链脂肪酸含量检测
精确称取50 mg样品于2 mL离心管中，加入0.2 mL 0.5%磷酸溶液，投入钢珠，于20 Hz下研磨10 min，涡旋混匀，加入500 μL甲基叔丁基醚，涡旋混匀，超声提取15 min，4 ℃、12 000 r/min离心10 min，取上清液进行GC-MS分析。
以甲基叔丁基醚为溶剂，先制备1 mg/mL混合标准品储备液，再稀释为0.1、0.2、0.5、1、2、10、20、50 mg/L的标准品系列溶液。
GC-MS分析条件：DB-FFAP色谱柱(30 m×0.25 mm×0.25 μm)，载气为氦气，流速1.0 mL/min。程序升温：90 ℃保持1 min，以25 ℃/min升至100 ℃，以20 ℃/min升至150 ℃并保持0.6 min，再以25 ℃/min升至200 ℃并保持0.5 min，最后以15 ℃/min升至230 ℃并保持1 min；采用单离子监测(single ion monitoring, SIM)模式分析。进样口温度和质谱传输线温度分别为200 ℃和230 ℃。
1.10 肠道菌群分析
采用Soil DNA Kit提取样本DNA，经0.8%琼脂糖凝胶电泳检测DNA完整性。使用引物338F (5′-ACTCCTACGGGAGGCAGCA-3′)和806R (5′-GGACTACHVGGGTWTCTAAT-3′)对16S rRNA基因V3-V4区进行PCR扩增。PCR反应体系(25 µL)：5×缓冲液5 µL，快速Pfu DNA聚合酶(5 U/µL) 0.25 µL，dNTPs (2.5 mmol/L) 2 µL，上、下游引物(10 µmol/L)各1 µL，DNA模板1 µL，ddH2O 14.75 µL。PCR反应条件：98 ℃预变性5 min；98 ℃变性30 s，53 ℃退火30 s，72 ℃延伸45 s，共25个循环；72 ℃终延伸5 min。借助Illumina NovaSeq 6000 SP测序系统，进行双端2×250 bp测序，获取基因序列信息。
1.11 统计学方法
使用SPSS 27.0软件进行统计分析，GraphPad Prism 9.5软件绘制图表，实验数据以mean±SD表示，组间比较采用单因素方差分析，结合Tukey事后检验，以P<0.05为差异有统计学意义。
2 结果与分析
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2.1 牛樟叶对酒精性肝病小鼠肝脏指数的影响
与正常组相比，模型组肝脏指数显著升高(P<0.05)。与模型组相比，牛樟叶中剂量组、牛樟叶高剂量组及水飞蓟宾组肝脏指数均显著降低(P<0.05)，表明给药后肝脏指数得到明显改善。牛樟叶低剂量组肝脏指数虽有降低，但与模型组相比差异无统计学意义(P>0.05)；而牛樟叶中剂量组与高剂量组均表现出显著改善(P<0.05)，提示牛樟叶对肝脏指数的影响可能存在一定的剂量依赖性，即随剂量增加其改善作用增强(图1)。
2.2 牛樟叶对酒精性肝病小鼠生化指标的影响
与正常组相比，模型组天冬氨酸氨基转移酶水平显著升高(P<0.05)。与模型组相比，各给药组均能显著降低天冬氨酸氨基转移酶水平(P<0.05)，其中牛樟叶高剂量组降幅最为明显，表明牛樟叶各剂量组均能有效降低天冬氨酸氨基转移酶水平，且以高剂量组效果最佳(图2A)。
与正常组相比，模型组丙氨酸氨基转移酶水平显著升高(P<0.05)，进一步证实酒精所致肝脏损伤。与模型组相比，水飞蓟宾组和牛樟叶各剂量组均显著降低丙氨酸氨基转移酶水平(P<0.05)，表明各剂量牛樟叶和水飞蓟宾均能有效抑制丙氨酸氨基转移酶升高，且牛樟叶高剂量组和水飞蓟宾组在改善该指标方面效果更为突出(图2B)。
综上所述，牛樟叶能有效降低酒精性肝病小鼠天冬氨酸氨基转移酶和丙氨酸氨基转移酶水平，改善肝功能，且该作用呈一定的剂量依赖性，其中以牛樟叶高剂量组的干预效果最为显著。
2.3 结肠组织病理学分析
与正常组相比，模型组小鼠结肠组织可见明显的黏膜结构破坏与病理损伤(红色箭头)，具体表现为绒毛排列稀疏、倒伏且高度降低，隐窝结构扭曲、与黏膜肌层分离且深度变浅，杯状细胞数量明显减少，固有层中有大量炎症细胞浸润。上述病理特征表明，酒精灌胃成功诱导了典型的肠道黏膜损伤。经水飞蓟宾及牛樟叶干预后，各给药组均表现出明显的病理改善，包括绒毛结构修复、隐窝形态恢复及炎症浸润减轻，说明牛樟叶能有效缓解酒精所致肠道损伤(图3)。
2.4 牛樟叶对酒精性肝病小鼠短链脂肪酸的影响
与正常组相比，模型组短链脂肪酸(乙酸、丙酸和丁酸等)含量均显著降低(P<0.05)。与模型组相比，各给药组短链脂肪酸水平均显著回升(P<0.05)。结果表明，牛樟叶可通过促进短链脂肪酸生成、调节菌群代谢对酒精性肝病小鼠发挥保护作用(表1)。
2.5 牛樟叶对酒精性肝病小鼠肠道菌群的影响
2.5.1 肠道微生物OTUs比较
OTUs分析结果显示(图4)，各组间肠道菌群结构存在显著差异。其中，牛樟叶高剂量组与水飞蓟宾组的特有OTUs数量最多，表明二者干预均能显著重塑菌群结构，增强其独特性。相比之下，模型组特有OTUs较少，提示酒精干预可能削弱了菌群的独特性，建模成功。此外，各组间共有OTUs极少，进一步印证了不同处理对肠道菌群组成的整体性影响。综上所述，牛樟叶与水飞蓟宾给药均能有效驱动肠道菌群发生特异性改变。
2.5.2 α多样性分析
与正常组相比，模型组α多样性及丰富度显著降低，表明酒精摄入对肠道菌群结构具有明显破坏作用。牛樟叶高剂量组肠道菌群α多样性指标较模型组显著恢复，说明牛樟叶高剂量组能够有效缓解酒精引起的菌群失衡，可能通过调节微生物群落结构促进肠道微生态的重建与稳态维持(图5)。
此外，牛樟叶高剂量组与水飞蓟宾组在各项α多样性指数上表现相当，提示牛樟叶在促进菌群恢复和维持肠道生态平衡方面具有与水飞蓟宾相似的作用。综上所述，牛樟叶对肠道菌群具有保护作用，并具有干预酒精诱导菌群紊乱的治疗潜力。
2.5.3 β多样性分析
β多样性能够反映群落结构的异质性程度。主坐标分析(principal coordinate analysis, PCoA)结果显示(图6A)，各组样本沿主坐标轴(Axis 1, Axis 2)呈明显分离，二者分别解释了17.2%与12.4%的群落变异。正常组与模型组样本在空间坐标中分布距离较远，表明酒精干预导致微生物群落结构发生显著改变；而牛樟叶高剂量组与水飞蓟宾组样本分布更接近正常组，提示牛樟叶治疗有助于酒精性肝病小鼠肠道菌群结构向正常状态恢复。
非度量多维标度(non-metric multidimensional scaling, NMDS)分析进一步验证了上述结论(图6B)，其应力值(stress=0.19)表明模型拟合优度处于可接受范围。在NMDS空间中各组样本各自独立聚类，其中牛樟叶高剂量组与正常组分布较为接近，而模型组样本呈相对分散的分布模式，与PCoA结果相互印证，共同说明牛樟叶能够改善酒精引起的肠道微生物群落紊乱。
综上所述，酒精摄入显著扰动了小鼠肠道微生物组成，而牛樟叶治疗能够有效逆转这一结构性失衡，促使菌群结构向正常状态恢复。该恢复效应可能与牛樟叶缓解酒精性肝病的内在机制有关，同时也进一步印证了肠道微生物群落在酒精性肝病发生与发展中的重要地位。
2.5.4 肠道菌群群落组成分析
门水平上各组肠道微生物群落结构组成显示(图7A)，与正常组相比，模型组中厚壁菌门(Firmicutes_D)和拟杆菌门(Bacteroidota)相对丰度均升高，F/B比值上升，表明酒精摄入干扰了基础菌群结构。经牛樟叶和水飞蓟宾干预后，厚壁菌门相对丰度下降，而拟杆菌门相对丰度升高，F/B比值下降，说明牛樟叶有助于恢复酒精引起的门水平菌群失衡。
科水平上各组肠道微生物组成变化显示(图7B)，与正常组相比，模型组中乳杆菌科(Lactobacillaceae)相对丰度升高，而毛螺菌科(Lachnospiraceae)相对丰度降低。经水飞蓟宾和牛樟叶治疗后，乳杆菌科相对丰度下降，毛螺菌科相对丰度回升，整体结构趋于正常，进一步证实牛樟叶对酒精扰动下菌群结构具有改善作用。
属水平上各组肠道微生物组成特征显示(图7C)，与正常组相比，模型组中乳杆菌属(Lactobacillus)和普雷沃氏菌属(Prevotella)相对丰度上升，而哺乳动物球菌属(Mammaliicoccus)相对丰度减少。经水飞蓟宾干预后，乳杆菌属相对丰度明显降低。牛樟叶高剂量组对乳杆菌属与变形菌属(Proteus)相对丰度虽有所改善但效果有限，提示牛樟叶对某些特定菌属的调控能力较弱，但仍有助于整体菌群结构的恢复。
综合门、科、属3个分类水平的结果可知，酒精摄入会显著改变小鼠肠道微生物组成，而牛樟叶治疗能够有效改善这一失衡状态，促进菌群整体恢复。该调节作用可能与牛樟叶缓解酒精性肝病的机制密切相关，同时也进一步表明肠道微生物群落在酒精性肝病发生与发展中具有重要地位。
2.5.5 关联网络分析
基于物种丰度数据构建的微生物共现网络揭示了类群间的相互关系及组间分布差异(图8)。在关联网络结构中，节点颜色代表不同分类，连线的粗细与颜色深浅反映类群间相关性强度；同时，图中还展示了不同微生物类群在各组样本中的具体分布模式。
在群落组成方面(图8A)，拟杆菌门和芽孢杆菌门在所有组别中均占据较高比例，为该生态系统中的优势菌门。在网络拓扑角色方面(图8B)，模块枢纽(module hubs)和网络枢纽(network hubs)分别在模块内部及整个网络中发挥核心作用；连接器(connectors)负责连接不同模块，而外围物种(peripherals)则处于网络边缘，功能较为独立。
综上所述，拟杆菌门与芽孢杆菌门在肠道微生态中占主导地位，而非优势类群(如假单胞菌等)在特定生理或扰动条件下可能承担关键功能。关联网络分析直观揭示了微生物类群间的复杂互作关系，为探索牛樟叶治疗酒精性肝病的作用机制提供了新视角，表明其可能通过调控肠道菌群结构与网络互作，在微生态层面发挥治疗作用。
2.5.6 肠道菌群代谢通路注释
基于KEGG数据库对盲肠内容物代谢产物进行功能注释的结果显示(图9)，主要富集的生物学功能集中于生物合成相关代谢通路，包括外源性物质生物降解与代谢、碳水化合物代谢、萜类与聚酮化合物代谢等核心模块。上述通路共同表明肠道微生物在宿主-环境界面中承担着关键物质转化功能，并从功能层面揭示了牛樟叶干预酒精性肝病的潜在机制，表明其可能通过调控菌群代谢网络，协同增强外源毒物清除能力、优化能量代谢平衡并调节免疫微环境，从多途径发挥治疗作用。
3 讨论与结论
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酒精性肝病的发生与肠道微生态失衡及有益代谢物短链脂肪酸的耗竭密切相关[13]。经牛樟叶干预可有效逆转这一病理状态，通过调节肠道菌群组成促进具有屏障修复与抗炎功能的短链脂肪酸生物合成，从而经“微生物-肠-肝轴”发挥肝脏保护作用。
酒精摄入可引起肠道菌群结构在多级分类水平上发生特征性紊乱[14]，门水平上表现为芽孢杆菌门/拟杆菌门比例升高，科水平上乳杆菌科异常增多而毛螺菌科减少。牛樟叶干预后能有效逆转酒精所致菌群结构失调，恢复微生态平衡，使整体菌群结构向正常组回归。这种抑制致病菌、促进有益菌恢复的双向调节作用与水飞蓟宾趋势一致，但针对乳杆菌属等特定菌属的调节效果存在差异，提示牛樟叶对特定菌群具有选择性调控作用。
短链脂肪酸作为肠道菌群代谢产生的关键活性物质，在维持肠道屏障完整性、调节免疫反应及肝脏代谢中发挥核心作用[15]，其含量可有效反映菌群代谢活性及机体微生态健康状况[16]。菌群结构的改善直接促进了短链脂肪酸等有益代谢产物的产量回升，是连接肠道与肝脏保护功能的关键环节[17]。模型组小鼠盲肠内容物中乙酸、丙酸、丁酸等短链脂肪酸含量均显著降低，经牛樟叶干预后短链脂肪酸水平显著恢复，这与毛螺菌科相对丰度回升高度吻合。由于短链脂肪酸(如丁酸)不仅能作为结肠细胞的主要能量来源、增强肠道屏障功能以减少有害物质易位，还能经循环系统调节全身及肝脏的免疫与炎症反应[18]，牛樟叶通过重塑菌群、促进短链脂肪酸生成，构建了一条从肠道微生态修复到“肠-肝轴”信号改善，最终实现肝损伤缓解的完整作用通路。
肠道菌群结构及其代谢产物短链脂肪酸的变化表明，牛樟叶对肠道菌群的调节机制可能涉及多重途径：作为益生元促进有益菌增殖[19]；调节肠道局部免疫，抑制促炎因子产生[20]；同时，通过恢复短链脂肪酸等代谢产物水平[21]，间接调控“肠-肝轴”信号通路，从而发挥肝保护效应。本研究通过多组学方法揭示了牛樟叶干预下“菌群-短链脂肪酸-肝损伤”三者之间的密切关联，为其保护作用提供了系统性的机制线索。不足之处在于，菌群变化与宿主表型之间的直接因果关系仍需通过粪菌移植[22]、无菌动物模型[23]或特定益生菌/短链脂肪酸补充实验[24]等进一步验证，这也是后续研究的重点方向。
基金
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  • 全国中药特色技术传承人才培训项目(T20234832005)
  • 广西中医药大学研究生教育创新计划(YCSY2025096)
文献
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参考文献 引证文献
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2026年第66卷第8期
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doi: 10.13343/j.cnki.wsxb.20260055
  • 接收时间：2026-01-20
  • 首发时间：2026-08-21
  • 出版时间：2026-08-04
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  • 收稿日期：2026-01-20
  • 录用日期：2026-02-27
基金
National Training Program for Inheriting Talents of Characteristic Technologies in Traditional Chinese Medicine(T20234832005)
全国中药特色技术传承人才培训项目(T20234832005)
Innovation Project of Guangxi Graduate Education of GXUCM(YCSY2025096)
广西中医药大学研究生教育创新计划(YCSY2025096)
作者信息
    1.广西中医药大学，广西 南宁
    2.柳州市中医医院(柳州市壮医医院)，广西 柳州
    3.广西中医药大学第一附属医院，广西 南宁
    4.柳州市妇幼保健院，广西 柳州

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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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