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Article(id=1322925455858335853, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923782029062261, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.06.002, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793339607442, onlineDateStr=2026-10-30, pubDate=1749484800000, pubDateStr=2025-06-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339607442, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339607442, creator=13701087609, updateTime=1793339607442, updator=13701087609, issue=Issue{id=1322923782029062261, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='6', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1749484800000', pubDateStr='2025-06-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339208371, creator='13701087609', updateTime=1793340556755, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322929437859258468, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923782029062261, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322929437859258469, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923782029062261, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=5, endPage=8, ext={EN=ArticleExt(id=1322925456059662446, articleId=1322925455858335853, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Exploring Mechanism of Guilu Erxian Glue（龟鹿二仙胶）in Preventing and Treating Postmenopausal Osteoporosis Based on “Gut Microbiota”, columnId=1256183423710970242, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=New Opinion of Ph.D. Supervisor, runingTitle=null, highlight=null, articleAbstract=
Objective

To observe the effect of Guilu Erxian Glue（龟鹿二仙胶，GLE）on the prevention and treatment of postmenopausal osteoporosis（PMO）in ovariectomized rats（OVX），and to explore the biological effects of gut microbiota.

Methods

The female SD rats were divided into 6 groups by random number table method，thus as Sham，OVX，vancomycin（KSS），GLE-L，GLE-M and GLE-H groups.After 1 week of modeling，the drug was administrated by gavage，and the sample was collected 8 weeks later.HE staining was used to detect bone structure，bone cell morphology and number.Bone microstructure was detected by microCT.Bone metabolism was detected by ELISA.16Sr DNA was used to analyze the V3-V4 region of the gut microbiota.

Results

The results of HE and microCT showed that different concentrations of GLE effectively increased bone trabecular percentage，restored bone microstructure，up-regulated the contents of serum ALP and PINP and down-regulated serumβ-CTX content（compared with OVX group，P<0.05）.OTU and PCoA analysis showed that GLE-M significantly restored the richness and diversity of intestinal flora and the composition of intestinal flora in OVX rats.At the phylum level，Firmicutes increased significantly in OVX group（compared with that in Sham group，P<0.01）and the flora abundance decreased significantly after GLE-M intervention（compared with that in OVX group，P<0.01）.At the genus level，the dominant bacteria in GLE group were lactobacillus，Klebsiella，Fusobacterium and so on.Eubacterium myxus increased in OVX group（compared with that in Sham group，P<0.01），and its abundance recovered significantly after GLE-M intervention（compared with that in OVX group，P>0.05）.

Conclusion

GLE may prevent postmenopausal osteoporosis by regulating gut microbiota.

, authors=Dongdong YU, Kuizhu CHEN, Qiang GUO, Haili GU, Long QIAO, Ye QIAO, authorsList=Dongdong YU, Kuizhu CHEN, Qiang GUO, Haili GU, Long QIAO, Ye QIAO, authorCompany=null, correspAuthors=null, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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=1322925463403888783, articleId=1322925455858335853, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=基于“肠道菌群”探讨龟鹿二仙胶防治绝经后骨质疏松症的机制, columnId=1256183426563096985, journalTitle=中华中医药学刊, columnName=博士导师新论, runingTitle=null, highlight=null, articleAbstract=
目的

观察龟鹿二仙胶（Guilu Erxian Glue，GLE）对去卵巢绝经后骨质疏松症（OVX）大鼠的防治作用，并探索肠道菌群在该过程中所发挥的生物效应。

方法

采用随机数字表法对SD雌性大鼠分组，共6组：假手术组（Sham），模型组（OVX），万古霉素组（KSS），龟鹿二仙胶低、中、高剂量组（GLE-L、GLE-M、GLE-H），造模1周后开始灌胃给药，8周后取材。HE染色检测骨结构及骨细胞形态、数量；micro CT检测骨微观结构；ELISA法检测骨代谢；16Sr DNA对肠道菌群的V3～V4区进行分析。

结果

HE、microCT结果显示，不同浓度的龟鹿二仙胶均有效提高了骨小梁百分比、恢复骨微观结构，上调了血清中ALP及PINP含量，下调了β-CTX含量（与模型组相比，P<0.05）。OTU和PCoA分析结果显示，GLE-M显著恢复了OVX大鼠肠道菌群丰富度与多样性及菌群构成；在门水平上，厚壁菌门在OVX组内明显增加（与Sham组比较，P<0.01），中药GLE-M干预后，其菌群丰度明显降低（与OVX组比较，P<0.01）。在属水平上，GLE组优势菌属为乳杆菌属（lactobacillus）、克雷伯菌属（Klebsiella）、梭杆菌属（Fusobacterium）等。黏液真杆菌属在OVX组内增加（与Sham组比较，P<0.01），GLE-M干预后，其丰富度明显恢复（与OVX组比较，P>0.05）。

结论

龟鹿二仙胶可能通过调控肠道菌群防治绝经后骨质疏松症。

, authors=于冬冬, 陈奎竹, 郭强, 顾海立, 乔隆, 乔野, authorsList=于冬冬, 陈奎竹, 郭强, 顾海立, 乔隆, 乔野, authorCompany=null, correspAuthors=null, authorNote=

于冬冬（1983-），男，黑龙江哈尔滨人，主任医师，博士研究生导师，博士，研究方向：中医药防治绝经后骨质疏松。

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于冬冬（1983-），男，黑龙江哈尔滨人，主任医师，博士研究生导师，博士，研究方向：中医药防治绝经后骨质疏松。

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于冬冬（1983-），男，黑龙江哈尔滨人，主任医师，博士研究生导师，博士，研究方向：中医药防治绝经后骨质疏松。

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Clin Lab，2020，66（12）：2443-2448., articleTitle=Associations of changes in serum inflammatory factors，MMP-3，25（OH）Dand intestinal flora with osteoporosis and disease activity in rheumatoid arthritis patients, refAbstract=null), Reference(id=1322925516432474380, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, doi=null, pmid=null, pmcid=null, year=2020, volume=40, issue=13, pageStart=2734, pageEnd=2736, url=null, language=null, rfNumber=［33］, rfOrder=32, authorNames=刘武, 崔潇镱, 滕健竣, journalName=中国老年学杂志, refType=null, unstructuredReference=刘武，崔潇镱，滕健竣，等. 二甲双胍联合鲑鱼降钙素治疗糖尿病性骨质疏松近期疗效及对肠道菌群、IGF-1和瘦素的影响［J］. 中国老年学杂志，2020，40（13）：2734-2736., articleTitle=二甲双胍联合鲑鱼降钙素治疗糖尿病性骨质疏松近期疗效及对肠道菌群、IGF-1和瘦素的影响, refAbstract=null)], funds=[Fund(id=1322925494575956191, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, awardId=82374480, language=CN, fundingSource=国家自然科学基金面上项目(82374480), fundOrder=null, country=null), Fund(id=1322925495301570785, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, awardId=zyzx1908, language=CN, fundingSource=辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金项目(zyzx1908), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1322925464699928719, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, xref=null, ext=[AuthorCompanyExt(id=1322925464733483152, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, companyId=1322925464699928719, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Liaoning University of Traditional Chinese Medicine，Shenyang 110847，Liaoning，China), AuthorCompanyExt(id=1322925465073221777, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, companyId=1322925464699928719, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=辽宁中医药大学，辽宁 沈阳 110847)])], figs=[ArticleFig(id=1322925484861948096, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=6nTwvDnuxRSx+v2cqGDfqA==, figureFileBig=8fLXs6qLAIc/hW8BnafSTg==, tableContent=null), ArticleFig(id=1322925484933251264, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图1, caption=大鼠股骨组织形态学（×10）, figureFileSmall=6nTwvDnuxRSx+v2cqGDfqA==, figureFileBig=8fLXs6qLAIc/hW8BnafSTg==, tableContent=null), ArticleFig(id=1322925485298155713, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=cD+yiKBUk2t7WSJZUAIyFg==, figureFileBig=7OFXmNULPMszrpAjty5AxQ==, tableContent=null), ArticleFig(id=1322925485637894338, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图2, caption=大鼠股骨组织三维成像图, figureFileSmall=cD+yiKBUk2t7WSJZUAIyFg==, figureFileBig=7OFXmNULPMszrpAjty5AxQ==, tableContent=null), ArticleFig(id=1322925487353364677, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=Jmx18n1mLHzArHWABz0cOg==, figureFileBig=G628l529V6y10bl957MxEg==, tableContent=null), ArticleFig(id=1322925487739240646, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图3, caption=韦恩图

注：z为Sham组；m为OVX组；y为GLE-M组；k为KSS组。

, figureFileSmall=Jmx18n1mLHzArHWABz0cOg==, figureFileBig=G628l529V6y10bl957MxEg==, tableContent=null), ArticleFig(id=1322925489408573640, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=+dOoEAzT8DKbI+XN6efgjA==, figureFileBig=XRZBJS4sOoFtDCnW3xMxmw==, tableContent=null), ArticleFig(id=1322925489496654025, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图4, caption=PCoA图, figureFileSmall=+dOoEAzT8DKbI+XN6efgjA==, figureFileBig=XRZBJS4sOoFtDCnW3xMxmw==, tableContent=null), ArticleFig(id=1322925489844781258, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=GF8z6saV5SEftUn0LD0kHQ==, figureFileBig=IwemjyPmzCP5tSdHmvRx/g==, tableContent=null), ArticleFig(id=1322925490310349003, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图5, caption=物种相对丰度柱状图（门水平）, figureFileSmall=GF8z6saV5SEftUn0LD0kHQ==, figureFileBig=IwemjyPmzCP5tSdHmvRx/g==, tableContent=null), ArticleFig(id=1322925490662670540, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=Ek015BR+6yRBd4OWP6n1cw==, figureFileBig=CuzN0U2bZmwKnVtE81BJfw==, tableContent=null), ArticleFig(id=1322925490725585101, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图6, caption=物种相对丰度柱状图（属水平）, figureFileSmall=Ek015BR+6yRBd4OWP6n1cw==, figureFileBig=CuzN0U2bZmwKnVtE81BJfw==, tableContent=null), ArticleFig(id=1322925490796888270, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=6cQusm+tB5i2SJ1/POfWaA==, figureFileBig=N9DjlEbQaF6JAPU4+hFbjw==, tableContent=null), ArticleFig(id=1322925490859802831, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图7, caption=进化分支图（LDA＝4）, figureFileSmall=6cQusm+tB5i2SJ1/POfWaA==, figureFileBig=N9DjlEbQaF6JAPU4+hFbjw==, tableContent=null), ArticleFig(id=1322925491140821200, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=Dz6xC9BOb/k1Sh4IjDqjhA==, figureFileBig=xbWokZaTW0dsz2fgYVDhPQ==, tableContent=null), ArticleFig(id=1322925491216318674, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=图8, caption=LDA值分布柱状图, figureFileSmall=Dz6xC9BOb/k1Sh4IjDqjhA==, figureFileBig=xbWokZaTW0dsz2fgYVDhPQ==, tableContent=null), ArticleFig(id=1322925491296010452, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n骨小梁平均厚度/（μm）骨小梁面积百分比/（%）
Sham组1050.213±8.13734.992±8.132
OVX组1041.131±8.014**15.554±6.917**
KSS组1041.975±8.07117.054±7.201
GLE-M组1047.063±8.209#28.705±5.301##
), ArticleFig(id=1322925491493142741, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=表1, caption=

大鼠股骨骨小梁改变情况（±s）

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组别n骨小梁平均厚度/（μm）骨小梁面积百分比/（%）
Sham组1050.213±8.13734.992±8.132
OVX组1041.131±8.014**15.554±6.917**
KSS组1041.975±8.07117.054±7.201
GLE-M组1047.063±8.209#28.705±5.301##
), ArticleFig(id=1322925491568640214, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n骨密度/（mm3）骨体积/（mm3）皮质骨密度/（mm3）皮质骨体积/（μm）骨小梁密度/（mm3）骨小梁体积/（mm3）
Sham组106 605.78##41.95##7 903.9#16.286 159.934.83##
OVX组105 149.87**34.63**7 761.02*15.496 041.790.71**
KSS组105 364.83#36.02#7 783.2415.856 049.852.02##
GLE-M组105 856.93#37.91#7 836.6816.246 054.823.71##
), ArticleFig(id=1322925491639943383, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=表2, caption=

大鼠股骨组织Micro CT扫描结果

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组别n骨密度/（mm3）骨体积/（mm3）皮质骨密度/（mm3）皮质骨体积/（μm）骨小梁密度/（mm3）骨小梁体积/（mm3）
Sham组106 605.78##41.95##7 903.9#16.286 159.934.83##
OVX组105 149.87**34.63**7 761.02*15.496 041.790.71**
KSS组105 364.83#36.02#7 783.2415.856 049.852.02##
GLE-M组105 856.93#37.91#7 836.6816.246 054.823.71##
), ArticleFig(id=1322925491933544666, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nALP含量/（U/L）
Sham组10303.742±3.483##
OVX组10145.371±2.546**
KSS组10150.442±2.879**
GLE-L组10150.442±2.879**
GLE-M组10230.525±3.197**##
GLE-H组10251.637±3.294**##
), ArticleFig(id=1322925492009042139, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=表3, caption=

大鼠血清ALP含量比较（±s）

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组别nALP含量/（U/L）
Sham组10303.742±3.483##
OVX组10145.371±2.546**
KSS组10150.442±2.879**
GLE-L组10150.442±2.879**
GLE-M组10230.525±3.197**##
GLE-H组10251.637±3.294**##
), ArticleFig(id=1322925492365557981, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nPINP含量β-CTX含量
Sham组103 549.392±137.853##370.548±60.597##
OVX组102 249.392±137.853##570.548±60.597##
KSS组102 304.933±192.868**515.756±64.845**
GLE-L组102 895.055±185.021**##421.257±64.875**##
GLE-M组103 165.163±163.543**##396.136±65.363**##
GLE-H组103 323.747±157.596**##386.937±65.843**##
), ArticleFig(id=1322925494043279582, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925455858335853, language=CN, label=表4, caption=

大鼠血清PINP、β-CTX含量比较（±s）

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组别nPINP含量β-CTX含量
Sham组103 549.392±137.853##370.548±60.597##
OVX组102 249.392±137.853##570.548±60.597##
KSS组102 304.933±192.868**515.756±64.845**
GLE-L组102 895.055±185.021**##421.257±64.875**##
GLE-M组103 165.163±163.543**##396.136±65.363**##
GLE-H组103 323.747±157.596**##386.937±65.843**##
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基于“肠道菌群”探讨龟鹿二仙胶防治绝经后骨质疏松症的机制
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于冬冬 , 陈奎竹 , 郭强 , 顾海立 , 乔隆 , 乔野
中华中医药学刊 | 博士导师新论 2025,43(6): 5-8
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中华中医药学刊 |博士导师新论 2025 , 43 (6) : 5 -8
基于“肠道菌群”探讨龟鹿二仙胶防治绝经后骨质疏松症的机制
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于冬冬（1983-），男，黑龙江哈尔滨人，主任医师，博士研究生导师，博士，研究方向：中医药防治绝经后骨质疏松。

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于冬冬（1983-），男，黑龙江哈尔滨人，主任医师，博士研究生导师，博士，研究方向：中医药防治绝经后骨质疏松。

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于冬冬, 陈奎竹, 郭强, 顾海立, 乔隆, 乔野
作者信息
  • 辽宁中医药大学，辽宁 沈阳 110847
作者简介:

于冬冬（1983-），男，黑龙江哈尔滨人，主任医师，博士研究生导师，博士，研究方向：中医药防治绝经后骨质疏松。

Exploring Mechanism of Guilu Erxian Glue（龟鹿二仙胶）in Preventing and Treating Postmenopausal Osteoporosis Based on “Gut Microbiota”
Dongdong YU, Kuizhu CHEN, Qiang GUO, Haili GU, Long QIAO, Ye QIAO
Affiliations
  • Liaoning University of Traditional Chinese Medicine，Shenyang 110847，Liaoning，China
出版时间: 2025-06-10 doi: 10.13193/j.issn.1673-7717.2025.06.002
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摘要
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目的

观察龟鹿二仙胶（Guilu Erxian Glue，GLE）对去卵巢绝经后骨质疏松症（OVX）大鼠的防治作用，并探索肠道菌群在该过程中所发挥的生物效应。

方法

采用随机数字表法对SD雌性大鼠分组，共6组：假手术组（Sham），模型组（OVX），万古霉素组（KSS），龟鹿二仙胶低、中、高剂量组（GLE-L、GLE-M、GLE-H），造模1周后开始灌胃给药，8周后取材。HE染色检测骨结构及骨细胞形态、数量；micro CT检测骨微观结构；ELISA法检测骨代谢；16Sr DNA对肠道菌群的V3～V4区进行分析。

结果

HE、microCT结果显示，不同浓度的龟鹿二仙胶均有效提高了骨小梁百分比、恢复骨微观结构，上调了血清中ALP及PINP含量，下调了β-CTX含量（与模型组相比，P<0.05）。OTU和PCoA分析结果显示，GLE-M显著恢复了OVX大鼠肠道菌群丰富度与多样性及菌群构成；在门水平上，厚壁菌门在OVX组内明显增加（与Sham组比较，P<0.01），中药GLE-M干预后，其菌群丰度明显降低（与OVX组比较，P<0.01）。在属水平上，GLE组优势菌属为乳杆菌属（lactobacillus）、克雷伯菌属（Klebsiella）、梭杆菌属（Fusobacterium）等。黏液真杆菌属在OVX组内增加（与Sham组比较，P<0.01），GLE-M干预后，其丰富度明显恢复（与OVX组比较，P>0.05）。

结论

龟鹿二仙胶可能通过调控肠道菌群防治绝经后骨质疏松症。

龟鹿二仙胶  /  肠道菌群  /  绝经后骨质疏松症
Objective

To observe the effect of Guilu Erxian Glue（龟鹿二仙胶，GLE）on the prevention and treatment of postmenopausal osteoporosis（PMO）in ovariectomized rats（OVX），and to explore the biological effects of gut microbiota.

Methods

The female SD rats were divided into 6 groups by random number table method，thus as Sham，OVX，vancomycin（KSS），GLE-L，GLE-M and GLE-H groups.After 1 week of modeling，the drug was administrated by gavage，and the sample was collected 8 weeks later.HE staining was used to detect bone structure，bone cell morphology and number.Bone microstructure was detected by microCT.Bone metabolism was detected by ELISA.16Sr DNA was used to analyze the V3-V4 region of the gut microbiota.

Results

The results of HE and microCT showed that different concentrations of GLE effectively increased bone trabecular percentage，restored bone microstructure，up-regulated the contents of serum ALP and PINP and down-regulated serumβ-CTX content（compared with OVX group，P<0.05）.OTU and PCoA analysis showed that GLE-M significantly restored the richness and diversity of intestinal flora and the composition of intestinal flora in OVX rats.At the phylum level，Firmicutes increased significantly in OVX group（compared with that in Sham group，P<0.01）and the flora abundance decreased significantly after GLE-M intervention（compared with that in OVX group，P<0.01）.At the genus level，the dominant bacteria in GLE group were lactobacillus，Klebsiella，Fusobacterium and so on.Eubacterium myxus increased in OVX group（compared with that in Sham group，P<0.01），and its abundance recovered significantly after GLE-M intervention（compared with that in OVX group，P>0.05）.

Conclusion

GLE may prevent postmenopausal osteoporosis by regulating gut microbiota.

Guilu Erxian Glue（龟鹿二仙胶）  /  gut microbiota  /  postmenopausal osteoporosis
于冬冬, 陈奎竹, 郭强, 顾海立, 乔隆, 乔野. 基于“肠道菌群”探讨龟鹿二仙胶防治绝经后骨质疏松症的机制. 中华中医药学刊, 2025 , 43 (6) : 5 -8 . DOI: 10.13193/j.issn.1673-7717.2025.06.002
Dongdong YU, Kuizhu CHEN, Qiang GUO, Haili GU, Long QIAO, Ye QIAO. Exploring Mechanism of Guilu Erxian Glue（龟鹿二仙胶）in Preventing and Treating Postmenopausal Osteoporosis Based on “Gut Microbiota”[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (6) : 5 -8 . DOI: 10.13193/j.issn.1673-7717.2025.06.002
正文
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骨质疏松症（osteoporosis，OP）是一种骨代谢性疾病，其特点是骨密度低下、骨量减少、骨微结构改变从而导致骨骼的脆性增加、骨折易感性提高[1]。绝经后女性由于体内雌激素的缺乏，从而诱发的骨质疏松又称绝经后骨质疏松症（postmenopausal osteoporosis，PMO）。全球约30%的50岁以上绝经后妇女患有PMO[2-3]。目前PMO的治疗以选择性雌激素受体调节剂、双膦酸盐类药物为主[4]，虽然具有一定疗效，但不良反应明显。中医药能够从整体进行辨证论治，历史悠久，前景广泛[5]。
PMO属中医学“骨枯”“骨痿”及“骨极”等疾病范畴[6]。《医经精义》曰：“肾藏精，精生髓，髓养骨，故骨者，肾之合也，髓者，精之所生也，精足则髓足，髓在骨内，足则骨强。”《素问·上古天真论篇》中描述绝经后妇女生理状态：“七七任脉虚，太冲脉衰少，天癸竭，地道不通，故形坏而无子也。”由此可见，PMO妇女肾的脏腑机能衰退，精少髓枯，骨失髓养，初为肾阴亏损，后累及肾阳气不化，久病则阴阳俱虚[7]，故“补肾益精、阴阳并补法”是PMO的重要防治方法。
龟鹿二仙胶出自《医便》，对肾阴阳两虚型OP疗效确切[8]。方中鹿角胶通督脉而补肾壮阳，龟板滋通任脉而滋补肾阴，配伍人参大补元气，枸杞子补肾益血，全方沟通任督二脉，阴阳相生相化，气固血充而达到气血阴阳互补之功效。研究表明肠道菌群及其代谢产物与PMO的发生与发展有着紧密的联系，其可通过介导肠道代谢产物、免疫和内分泌系统等因素影响骨稳态水平[9-11]。基于以上理论，本研究采用去卵巢（OVX）雌性大鼠作为绝经后骨质疏松症模型，在观察龟鹿二仙胶对PMO防治作用的同时，检测大鼠肠道菌群的差异，探索其潜在机制。
1　材料与方法
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1.1　材料
1.1.1　实验动物
60只7～8周龄SPF级雌性SD大鼠于辽宁长生生物技术股份有限公司购入，体质量为（200±20）g，许可证号：SCXK（辽）2020-0001，饲养于辽宁中医药大学动物实验中心，实验单位许可证编号：SYXK（辽）2013-0009。
1.1.2　主要实验药品
龟鹿二仙胶（龟板489 g，鹿角胶489 g，人参489 g，枸杞733 g），购于辽宁省沈阳市康多辽河药房。盐酸万古霉素（上海研生实业有限公司）。
1.1.3　其他主要试剂
HE染色试剂盒、4%多聚甲醛溶液、EDTA脱钙液（索莱宝，北京）；ELISA试剂盒ALP、PINP、β-CTx（伊莱瑞特，武汉）。
1.1.4　主要实验设备
石蜡切片机（徕卡显微系统，上海）；生物组织包埋机（徕卡，德国）；超纯水机（Millipore，美国）；全自动脱水机（徕卡，德国）；电子光学显微镜（尼康，日本）；Micro CT扫描机（ZKKSMCT-Sharp-I，广州中科恺盛）；超低温冰箱（中科美菱，合肥）。
1.2　方法
1.2.1　实验动物的饲养及处理
（1）大鼠饲养：大鼠于辽宁中医药大学动物实验中心饲养，经紫外线灯照射消毒，恒温、恒湿饲养1周，期间保证大鼠摄入水粮自由。
（2）大鼠分组：将60只大鼠按随机分组法随机分为6组，每组10只：假手术组（Sham组）；模型组（OVX组）；抗生素组（KSS组）；龟鹿二仙胶低剂量组（GLE-L）；龟鹿二仙胶中剂量组（GLE-M）；龟鹿二仙胶高剂量组（GLE-H）。
（3）造模：术前所有大鼠禁食水12 h。麻醉剂选择3%戊巴比妥纳溶液，现配现用。按照0.1 mL/100 g的剂量标准配置麻醉剂对大鼠行腹腔注射麻醉，将麻醉后的大鼠俯卧固定于实验台，消毒、备皮后，在股骨近端连线与后正中线交点处，切开大约2 cm纵行切口，摘除双侧卵巢组织后止血、缝合及消毒。假手术组取等量的皮下脂肪组织。
（4）大鼠灌胃给药：依据第4版《药理实验方法学》中列出的换算公式，得出龟鹿二仙胶给药浓度为1.7 g/mL。Sham组大鼠灌胃等体积饮用水，KSS组灌胃等体积含万古霉素0.5 g/L的饮用水，OVX组：4.03 g/kg；GLE-L组：2.015 g/kg；GLE-M组：4.03 g/kg；GLE-H组：8.06 g/kg。SD大鼠在OVX术后观察7 d，7 d后开始每日灌胃给药，持续8周。
（5）大鼠血清、骨组织标本收集：实验结束时，将大鼠麻醉成功后，仰卧固定于实验台，经腹主动脉采血后注入采血管，离心取上清液置于EP管中，做好组别标记，于-80℃冰箱中冷冻备用。取大鼠两侧股骨，去除附着组织，用生理盐水漂洗干净，外包锡纸，于-80℃冰箱冷冻备用。
（6）粪便收集：无菌条件下取出大鼠结肠内的粪便组织，约0.2 g，置于1.5 mL的EP管内，做好组别标记，于-80℃冰箱中冷冻备用。
（7）大鼠股骨组织的固定、脱钙、包埋处理：用4%多聚甲醛溶液将骨组织固定5 d后，放入15%乙二胺四乙酸溶液脱钙4周左右，以大头针可轻松刺入股骨确定脱钙成功后进行包埋。
1.2.2　指标检测方法
（1）骨组织切片制作：将蜡块放入4℃冰箱中冷却20 min后取出，固定于切片机，6μm厚度进行切片，将切片置于摊片机中，待切片充分展开后使用黏附载玻片捞取组织切片，置于生物组织烤片机上烤片，待充分干燥后备用。
（2）HE染色法：将骨组织切片进行脱蜡复水、经高浓度到低浓度乙醇浸泡处理，参照试剂盒说明书进行HE染色再脱水透明后封片。
（3）Micro CT检测骨结构：用4%多聚甲醛溶液固定股骨标本24 h后，将股骨标本固定在Micro CT载物台上扫描，角度为360°，分辨度为10μm，以股骨近端1.5 mm区域作为ROI（感兴趣区域），在主机上建立骨小梁3D图像后进行软件分析，主要检测参数包括：骨体积、骨密度、皮质骨体积、皮质骨厚度、骨小梁体积、骨小梁厚度。
（4）ELISA法检测血清骨代谢指标：提前20 min在室温下取出ELISA试剂盒，提前15 min打开酶标仪，取出大鼠血清上清液，严格按照说明书指示进行，检测大鼠血清中骨代谢物ALP、PINP及β-CTX水平。
（5）大鼠肠道菌群检测：报送至北京诺禾致源生物科技有限公司，采用16SrRNA基因测序技术进行肠道菌群分析。
1.3　统计学方法
采用SPSS13.0版统计软件分析，两组间比较用计量资料t检验，多组间比较采用ANOVA方法，数据以均数±标准差（）表示，P<0.05差异有统计学意义。
2　结果
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2.1　HE染色结果
Sham组骨小梁连接紧密、排列整齐密集、结构呈网状；OVX组骨小梁明显变细、排列稀疏紊乱、断裂、大部分呈非网状结构；万古霉素组骨小梁略粗、有断裂、网状结构不明显；GLE-M组的骨小梁明显增粗、可见网状结构、排列致密整齐。见插页Ⅰ图1。
OVX组大鼠股骨平均骨小梁厚度、骨小梁面积百分比显著降低（P<0.01，与Sham组比较）；KSS组平均骨小梁厚度、骨小梁面积百分比无明显改变（P>0.05，与OVX组相比）；GLE-M 组平均骨小梁厚度、骨小梁面积百分比显著提高（P<0.01，与OVX组相比）。见表1。
2.2　Micro CT检测结果
骨组织骨体积由高到低排序为Sham组>GLE-M组>KSS组>OVX组；骨密度由高到低排序：Sham组>GLE-M组>KSS组>OVX组；皮质骨体积检测结果显示：各组间对比差异无明显统计学意义（P>0.05）；皮质密度检测结果显示：OVX组明显降低（P<0.05，与Sham组相比），GLE-M组改变不明显（P>0.05，OVX组）；骨小梁体积：Sham组>GLE-M组>KSS组>OVX组。骨小梁密度：Sham组>GLE-M组>KSS组>OVX组，GLE-M组与OVX组差异无明显统计学意义（P>0.05），见插页Ⅰ图2、表2。
2.3　骨代谢生化指标结果
2.3.1　血清ALP含量
OVX组、KSS组ALP含量明显减少（P<0.01，与Sham组相比）；GLE各剂量组随剂量的增大ALP含量随之升高，GLE-H组ALP的提升效果最明显（P<0.01，与OVX组相比）。见表3。
2.3.2　大鼠血清PINP及β-CTX含量
OVX组、KSS组PINP含量明显减少，β-CTX含量明显增加（P<0.01，与Sham组比较）；GLE各剂量组随着剂量的增大呈现出PINP含量随之升高、β-CTX相继降低，其中GLE-M组PINP的提升、β-CTX的减低效果最为明显（P<0.01，与Sham组相比较）。见表4。
2.4　龟鹿二仙胶对去卵巢骨质疏松症大鼠肠道菌群影响
2.4.1　OTU分析结果
按照97%序列相似性，聚类生成操作性分类单元（Operational Taxonomic Unit，OUT），韦恩（venn）图中每个圈代表一组样品，圈和圈重叠部分的数字代表样组之间共有的OTUs个数，没有重叠部分的数字代表样组的特有OTUs个数。结果显示：OVX组的OTU数明显减少（与Sham组比较，P<0.01），GLE-M组的OTU数有所恢复，但未达到正常水平（与OVX组比较，P<0.01），KSS组的OTU数与GLEM组相似（P>0.01）。见插页Ⅰ图3。
2.4.2　PCoA分析结果
主坐标分析（Principal Co-ordinates Analysis，PCoA），是基于Weighted Unifrac距离和Unweighted Unifrac距离来进行PCoA分析，选取贡献率最大的主坐标组合进行作图展示。如果样品距离越接近，表示物种组成结构越相似，群落差异很大的样品则会远远分开。如插页Ⅰ图4所示：可见OVX组、Sham组与GLE-M组肠道菌群结构相似，KSS组其他3组肠道菌群结构差异较大。
2.4.3　菌群鉴定与物种丰度
（1）在门水平上，菌群相对丰度结果显示（见插页Ⅱ图5），厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidetes）、变形菌门（Proteobacteria）、梭杆菌门（Fusobacterium）物种相对丰度存在显著差异，尤其是厚壁菌门，其在OVX组内明显增加（与Sham组比较，P<0.01），中药GLE-M干预后，其菌群丰度明显降低（与OVX组比较，P<0.01）。而拟杆菌门在OVX组内减低（与Sham组比较，P<0.05），GLEM干预后，减低更加明显（与OVX组比较，P<0.01）。变形菌门在OVX组内减低（与Sham组比较，P<0.01），GLE-M干预后，其丰度明显恢复（与OVX组比较，P>0.05）。
（2）在属水平上，菌群相对丰度结果显示（见插页Ⅱ图6），OVX组狭义梭菌属1（Clostridium sensu stricto 1）、苏黎世杆菌属（Turicibacter）、埃希氏菌属（Escherichia-shigella）物种相对丰度降低（与Sham组相比）；GLE-M组为乳杆菌属（lactobacillus）、克雷伯菌属（Klebsiella）、梭杆菌属（Fusobacterium）、埃希氏菌属（Escherichia）物种相对丰度升高；经黏液真杆菌属（Blautia）、狭义梭菌属1（Clostridium sensu stricto 1）、苏黎世杆菌属（Turicibacter）、罗氏菌属（Roseburia）物种相对丰度降低（与OVX组比较）。其中经黏液真杆菌属在OVX组内增加（与Sham组比较，P<0.01），GLE-M干预后，其丰度明显恢复（与OVX组比较，P>0.05）。
2.4.4　进化分支
进化分支图（见插页Ⅱ图7）显示了GLEM组在不同层级的富集状态，在纲水平为芽孢杆菌纲（Bacillus）、厚壁菌纲（Firmicutes）、梭杆菌纲（Fusobacterium）等；在目水平为乳杆菌目（Lactobacillus）、韦荣氏球菌目（Veillonellales_Selenomon）、梭杆菌目（Fusobacterium）等；在科水平为乳杆菌科（Lactobacillus）、韦荣氏球菌科（Veillonellales_Selenomon）、梭杆菌科（Fusobacterium）等。
2.4.5　组间差异菌群分析
LDA值分布柱状图（见插页Ⅲ图8）反映了各组具有显著差异的物种对其组间差异的影响大小。GLE-M组对比其余各组相对丰度较高的差异物种在门水平上为变形菌门（Proteobacteria）、梭杆菌门（Fusobacterium）、疣微菌门（Verrucomicrobiota）；在属水平为乳杆菌属（lactobacillus）、克雷伯菌属（Klebsiella）、梭杆菌属（Fusobacterium）、埃希氏菌属（Escherichia）等。
3　讨论
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PMO是常见的骨代谢疾病，严重地影响患者健康和生活质量。PMO目前尚无特效的治疗药物，临床使用的抗PMO药物存在多种不良反应（唑来膦酸钠的发热及骨痛，雌激素替代疗法增加子宫内膜癌的风险）及患者耐受程度差等问题[12]。越来越多的研究表明，肠道菌群可通过调节肠屏障、营养吸收和免疫调节等改善骨代谢[13-17]。因此课题组基于大鼠的肠道菌群进行动态分析，深入挖掘龟鹿二仙胶的抗PMO机制。
PMO的主要病机为肾精亏虚，亦与肝、脾等脏腑密切相关[18-19]。基于此病机，“补肾益精法”被认为是防治骨质疏松症的主要方法[20]。传统方剂龟鹿二仙胶有补益气血、填精益髓的功效，李士材将其特点概括为“一阴一阳，无偏胜之忧。入气入血，有和平之美”，以龟鹿二仙胶补肾填精，亦是治病求其本的表现。研究发现龟鹿二仙胶主要成分为胶原蛋白、脂肪酸、人参皂苷、鹿茸多肽等[21]。龟鹿二仙胶也具有抗炎、抗氧化等作用，能减轻患者疼痛，提高骨密度[26-27]。本研究结果显示，龟鹿二仙胶有助于提高骨密度，改善骨微结构，上调血清中ALP及PINP含量、下调了β-CTX含量水平，促进骨生成。
研究发现，人参、鹿茸、枸杞的活性成分人参皂苷、人参多糖、鹿茸多肽和枸杞多糖可以调节肠道菌群的丰度以及微生物群落的功能[22-25]。我们的结果显示，假手术组的肠道菌群多样性和丰度明显高于模型组，提示模型组大鼠骨质流失的发生可能与肠道菌群失衡相关。应用龟鹿二仙胶后，模型组大鼠肠道菌群组成、菌群的丰富度及多样性也相对得到了提升，接近假手术组。在门、属两水平物种相对丰度显示，发生变化的主要菌群为厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria），尤其是厚壁菌门，与假手术组相比较，模型组厚壁菌门丰度增加，而在龟鹿二仙胶中剂量组中厚壁菌门丰度降低，此结果与有关的报道相似[28]。YAN J等[29]发现厚壁菌门可通过免疫调节影响骨代谢。研究[30-31]表明，在骨质疏松症大鼠的肠道中，厚壁菌门的丰度显著提高，可通过降低其在肠道中的丰度来进行骨质疏松症治疗。LefSe分析中龟鹿二仙胶中剂量组中乳杆菌（Lactobacillus）丰度均明显高于其他3组。乳杆菌是一种在生物肠道内广泛存在的益生菌，其与OP的发生密切相关 [32-33]。
综上所述，龟鹿二仙胶可能通过肠道菌直接或间接影响骨代谢，进而达到防PMO的目的。肠道菌群是处于动态的群体，其复杂的生理调控机制仍需进一步探索。后续课题组将继续深入探究龟鹿二仙胶通过肠道菌群防治PMO的潜在机制，为其应用与临床防治PMO提供更加深入的理论依据。
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  • 国家自然科学基金面上项目(82374480)
  • 辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金项目(zyzx1908)
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doi: 10.13193/j.issn.1673-7717.2025.06.002
  • 首发时间：2026-10-30
  • 出版时间：2025-06-10
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国家自然科学基金面上项目(82374480)
辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金项目(zyzx1908)
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    辽宁中医药大学，辽宁 沈阳 110847
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2种不同金属材料的力学参数
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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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