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Article(id=1315688508115763274, tenantId=1146029695717560320, journalId=1315121837170065538, issueId=1315688435411694395, articleNumber=null, orderNo=null, doi=10.16368/j.issn.1674-8999.2025.12.413, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1749830400000, receivedDateStr=2025-06-14, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1791614184645, onlineDateStr=2026-10-10, pubDate=1765296000000, pubDateStr=2025-12-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1791614184645, onlineIssueDateStr=2026-10-10, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1791614184645, creator=13701087609, updateTime=1791614184645, updator=13701087609, issue=Issue{id=1315688435411694395, tenantId=1146029695717560320, journalId=1315121837170065538, year='2025', volume='40', issue='12', pageStart='2487', pageEnd='2712', issueExtLink='null', onlineDate='null', pubDate='1765296000000', pubDateStr='2025-12-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1791614167309, creator='13701087609', updateTime=1791616084449, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1315696476567744680, tenantId=1146029695717560320, journalId=1315121837170065538, issueId=1315688435411694395, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1315696476571938985, tenantId=1146029695717560320, journalId=1315121837170065538, issueId=1315688435411694395, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2650, endPage=2660, ext={EN=ArticleExt(id=1315688508468084811, articleId=1315688508115763274, tenantId=1146029695717560320, journalId=1315121837170065538, language=EN, title=Action Mechanism of Acupotomy Intervention in Rabbit Knee Osteoarthritis Cartilage Based on Transcriptomics, columnId=1315688455783428135, journalTitle=Acta Chinese Medicine, columnName=Experimental Study, runingTitle=null, highlight=null, articleAbstract=

Objective:To explore the molecular mechanism of acupotomy intervention in knee osteoarthritis（KOA）based on transcriptome sequencing technology.Methods:Nine New Zealand rabbits were randomly divided into a blank group（3 rabbits）and a KOA modeling group（6 rabbits）.The modeling group was subjected to a modified Videman method fixation for6 weeks to establish the KOA model.After removing the fixation device，the rabbits were further randomly divided into a model group and an acupotomy group（3 rabbits each）according to Lequesne scores and intervened for 4 weeks.After the intervention，behavioral assessments were conducted，and transcriptome sequencing，Gene Ontology（GO），and Kyoto Encyclopedia of Genes and Genomes（KEGG）analyses were performed on the rabbit knee cartilage.Results:（1）After the intervention，compared with the model group，the Lequesne scores in the acupotomy group were significantly reduced（P＜0.01）；（2）Compared with the blank group，a total of 1 055 differentially expressed genes（DEGs）were identified in the model group.GO biological process analysis mainly involved functions such as ″inflammatory response″，″immune response″，and″extracellular matrix organization″.KEGG Level 2 analysis showed that the model group was mainly enriched in pathways related to ″infectious diseases″，″signal transduction″，and″immune system″.In addition，the upregulated DEGs in the model group were enriched in pathways such as″cytokine-cytokine receptor interaction″and″phagosome″，while the downregulated DEGs were enriched in ″cancer pathways″and ″axon guidance″. Compared with the model group，312 differentially expressed genes were identified in the acupotomy group.GO analysis indicated that these genes primarily involved functions such as″inflammatory response″，″tumor necrosis factor response″，and ″semaphorin-plexin signaling pathway″.KEGG Level 2 analysis showed enrichment mainly in pathways related to ″signal transduction″，″infectious diseases″，and ″immune system″.Moreover，downregulated DEGs in the acupotomy group were enriched in ″cytokine-cytokine receptor interaction″，″phagosome″，and ″osteoclast differentiation″，while upregulated DEGs were enriched in ″axon guidance″and ″protein digestion and absorption″.（3）qRT-PCR validation showed that acupotomy intervention significantly reduced the mRNA expression of CCL2，CCL7，and IL34. Conclusion:The molecular mechanism of acupotomy intervention in KOA mainly involves functions related to joint cartilage ″signal transduction″，″immune-inflammatory response″，and pathways such as ″cytokine-cytokine receptor interaction″，″phagosome″，and ″axon guidance″.

, authors=Xiaowei SHI, Wenjing YU, Dongfeng WANG, Meng XU, Dapeng LIU, Zeming YAN, Zhiduo ZHANG, Shenghua HU, Shufeng SHI, authorsList=Xiaowei SHI, Wenjing YU, Dongfeng WANG, Meng XU, Dapeng LIU, Zeming YAN, Zhiduo ZHANG, Shenghua HU, Shufeng SHI, authorCompany=null, correspAuthors=Shufeng SHI, 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=1315688510686871650, articleId=1315688508115763274, tenantId=1146029695717560320, journalId=1315121837170065538, language=CN, title=基于转录组学探讨针刀干预对膝骨关节炎兔关节软骨的作用机制, columnId=1315688455959588905, journalTitle=中医学报, columnName=实验研究, runingTitle=null, highlight=null, articleAbstract=

目的：基于转录组学测序技术探讨针刀干预膝骨关节炎（knee osteoarthritis，KOA）的分子机制。方法：9只新西兰兔随机分为空白组（3只）及KOA造模组（6只），造模组采用改良Videman法固定6周以建立KOA模型，拆除固定装置后根据Lequesne评分再随机分为模型组和针刀组（每组3只），干预4周。干预结束后进行行为学评价，并对兔膝关节软骨进行转录组学测序、基因本体论（Gene Ontology，GO）和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）分析。结果：（1）干预后，与模型组相比，针刀组Lequesne评分显著降低（P＜0.01）；（2）与空白组相比，模型组共筛选出1 055条差异表达基因（differentially expressed genes，DEGs），GO分析生物过程主要涉及“炎症反应”“免疫反应”和“细胞外基质组织”等功能，KEGG Level2分析显示模型组DEGs主要参与“感染性疾病”“信号转导”“免疫系统”等通路；模型组表达上调的DEGs主要参与“细胞因子相互作用”“吞噬体”等通路，而表达下调的DEGs主要参与“癌症通路”“轴突导向”等通路。与模型组相比，针刀组共筛选出312条DEGs，GO分析结果显示，这些基因主要涉及“炎症反应”“肿瘤坏死因子反应”“semaphorin-plexin信号通路”等功能，KEGG Level2分析显示其主要参与“信号转导”“感染性疾病”及“免疫系统”等通路；针刀组表达下调的DEGs参与“细胞因子相互作用”“吞噬体”“破骨细胞分化”等通路，表达上调的DEGs参与“轴突导向”“蛋白质消化和吸收”等通路。（3）进行qRT-PCR验证，结果显示：针刀干预显著降低CCL2、CCL7、IL34 mRNA表达。结论：针刀干预KOA的分子机制主要涉及关节软骨“信号转导”“免疫-炎症”等功能以及“细胞因子受体相互作用”“吞噬体”“轴突引导”等通路。

, authors=史晓伟, 于文静, 王东峰, 徐萌, 刘大鹏, 闫泽明, 张治多, 胡胜华, 史术峰, authorsList=史晓伟, 于文静, 王东峰, 徐萌, 刘大鹏, 闫泽明, 张治多, 胡胜华, 史术峰, authorCompany=null, correspAuthors=史术峰, authorNote=

史晓伟（1986 -），男，河南济源人，医学博士，副研究员，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。

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史术峰，男，黑龙江孙吴人，医学博士，主任医师，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。E-mail:
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史晓伟（1986 -），男，河南济源人，医学博士，副研究员，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。

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史晓伟（1986 -），男，河南济源人，医学博士，副研究员，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。

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Cell Biol Int, 2023, 47(5):954-968., articleTitle=BML-111 inhibits osteoclast differentiation by suppressing the MAPK and NF-κB pathways,alleviating deterioration of the knee joints in a CIA rat model, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1315688510980472931, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, xref=null, ext=[AuthorCompanyExt(id=1315688510988861540, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, companyId=1315688510980472931, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=The Third Affiliated Hospital to Beijing University of Chinese Medicine, Beijing China 100029), AuthorCompanyExt(id=1315688510997250149, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, companyId=1315688510980472931, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=北京中医药大学第三附属医院，北京 100029)])], figs=[ArticleFig(id=1315688515837477023, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=8bxImwQbwUcUs5KM6nnTpw==, figureFileBig=L8ql4+9GV/5AHQWTRBQIyQ==, tableContent=null), ArticleFig(id=1315688515917168800, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图1, caption=与空白组相比模型组DEGs图

注：A：模型组与空白组DEGs聚类分析热图；control代表空白组，model代表模型组；红色代表高表达，蓝色代表低表达。B：模型组与空白组DEGs火山图；绿色代表高表达，红色代表低表达，灰色代表无差异。

, figureFileSmall=8bxImwQbwUcUs5KM6nnTpw==, figureFileBig=L8ql4+9GV/5AHQWTRBQIyQ==, tableContent=null), ArticleFig(id=1315688516080746657, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=l6gaSsRLmBRiIMI4XyXtmg==, figureFileBig=mXLjxRYYqgtLt6Gn/lBuSQ==, tableContent=null), ArticleFig(id=1315688516152049826, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图2, caption=与模型组相比针刀组DEGs图

注：A：针刀组与模型组DEGs聚类分析热图；Dao代表针刀组，model代表模型组；红色代表高表达，蓝色代表低表达。B：针刀组与模型组DEGs火山图；绿色代表低表达，红色代表高表达，灰色代表无差异。

, figureFileSmall=l6gaSsRLmBRiIMI4XyXtmg==, figureFileBig=mXLjxRYYqgtLt6Gn/lBuSQ==, tableContent=null), ArticleFig(id=1315688516210770083, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=Gkug8boc0umPaEdeMEDFvQ==, figureFileBig=ux8KuHeqndwPEgBM8+2OTg==, tableContent=null), ArticleFig(id=1315688516269490340, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图3, caption=模型组vs空白组DEGs GO富集分析（TOP 30 GO Term）, figureFileSmall=Gkug8boc0umPaEdeMEDFvQ==, figureFileBig=ux8KuHeqndwPEgBM8+2OTg==, tableContent=null), ArticleFig(id=1315688516336599205, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=jbzX8P1pk1joaT637b6/Ug==, figureFileBig=JI7VsBqwAoAJJdffdLfbvw==, tableContent=null), ArticleFig(id=1315688516403708070, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图4, caption=针刀组vs模型组DEGs GO富集分析（TOP 30 GO Term）, figureFileSmall=jbzX8P1pk1joaT637b6/Ug==, figureFileBig=JI7VsBqwAoAJJdffdLfbvw==, tableContent=null), ArticleFig(id=1315688516466622631, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=PoVLlQ5B4+nYlrAqxsZZNQ==, figureFileBig=yf3GzcKpi2oCQRn5PtzfNA==, tableContent=null), ArticleFig(id=1315688516533731496, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图5, caption=模型组vs空白组DEGs KEGG通路富集分析

注：A：模型组与空白组DEGs KEGG通路富集分析前20分布图（Enrichment top 20）；B：模型组与空白组DEGs与所有基因在KEGG Level2水平分布比较图（KEGG Pathway Classification）。

, figureFileSmall=PoVLlQ5B4+nYlrAqxsZZNQ==, figureFileBig=yf3GzcKpi2oCQRn5PtzfNA==, tableContent=null), ArticleFig(id=1315688516617617577, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=WBu8Gg+e4uAw6HSpQRw6fQ==, figureFileBig=1YSgo3nUTEIznfbtdl6MCQ==, tableContent=null), ArticleFig(id=1315688516697309354, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图6, caption=模型组vs空白组DEGs表达升高/降低KEGG富集分析图（Enrichment top 20）

注：A：模型组vs空白组DEGs表达升高KEGG通路富集分析前20分布图；B：模型组vs空白组DEGs表达降低KEGG通路富集分析前20分布图。

, figureFileSmall=WBu8Gg+e4uAw6HSpQRw6fQ==, figureFileBig=1YSgo3nUTEIznfbtdl6MCQ==, tableContent=null), ArticleFig(id=1315688516764418219, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=If7LFg+g3lrQgbsKuPwnSw==, figureFileBig=i23tkxYvavwVEU2FCskQpg==, tableContent=null), ArticleFig(id=1315688516839915692, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图7, caption=针刀组vs模型组DEGs KEGG通路富集分析

注：A：针刀组与模型组DEGs KEGG通路富集分析前20分布图（enrichment top 20）；B：针刀组与模型组DEGs与所有基因在KEGG Level2水平分布比较图（KEGG Pathway Classification）。

, figureFileSmall=If7LFg+g3lrQgbsKuPwnSw==, figureFileBig=i23tkxYvavwVEU2FCskQpg==, tableContent=null), ArticleFig(id=1315688516907024557, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=QMkXH2XfzYECKhl2EL2kUw==, figureFileBig=+tBSCaxI/RtgR0r77wLLqg==, tableContent=null), ArticleFig(id=1315688516999299246, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=图8, caption=针刀组vs模型组DEGs表达降低/升高KEGG富集分析图（enrichment top 20）

注：A：针刀组vs模型组DEGs表达降低KEGG通路富集分析前20分布图；B：针刀组vs模型组DEGs表达升高KEGG通路富集分析前20分布图。

, figureFileSmall=QMkXH2XfzYECKhl2EL2kUw==, figureFileBig=+tBSCaxI/RtgR0r77wLLqg==, tableContent=null), ArticleFig(id=1315688517066408111, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
基因前向引物（5'-3'）反向引物（5'-3'）
CCL2AGCCAACTCTCACTGAAGCCAACTGTGAACAACAGGCCCA
CCL7AAGCCCTGAAGACAGATGCCCCCCTTAGGACCGTAGTCCA
IL34TGAGCTGCTGTAGCTTGCATCTCTTGGAATCTGGGGCTGG
GADPHGGCATCCTGGGCTACACTGAGGAGTGGGTGTCGCTGTTG
), ArticleFig(id=1315688517158682800, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=表1, caption=

引物信息

, figureFileSmall=null, figureFileBig=null, tableContent=
基因前向引物（5'-3'）反向引物（5'-3'）
CCL2AGCCAACTCTCACTGAAGCCAACTGTGAACAACAGGCCCA
CCL7AAGCCCTGAAGACAGATGCCCCCCTTAGGACCGTAGTCCA
IL34TGAGCTGCTGTAGCTTGCATCTCTTGGAATCTGGGGCTGG
GADPHGGCATCCTGGGCTACACTGAGGAGTGGGTGTCGCTGTTG
), ArticleFig(id=1315688517221597361, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
时间n模型组针刀组t值P值
干预前36.33 ±0.576.00 ±0.001.000.374
干预后35.67 ±0.573.33 ±0.57*4.950.008
), ArticleFig(id=1315688517292900530, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=CN, label=表2, caption=

各组干预前后Lequesne评分比较

, figureFileSmall=null, figureFileBig=null, tableContent=
时间n模型组针刀组t值P值
干预前36.33 ±0.576.00 ±0.001.000.374
干预后35.67 ±0.573.33 ±0.57*4.950.008
), ArticleFig(id=1315688517355815091, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
样品Raw reads/MbRaw bases/GbClean reads/MbClean bases/GbQ30/%GC/%Total readsTotal mapped reads（%）
空白150.367.5545.696.5290.7054.764568823241659835（91.18）
空白255.448.3252.407.5693.4154.105240423648549636（92.64）
空白352.057.8147.816.8391.6652.824781088443834282（91.68）
模型152.047.8148.707.0192.8752.594869794444530255（91.44）
模型253.618.0449.517.0892.2353.764951423445501233（91.89）
模型350.967.6445.746.5490.5154.554573905442532703（92.98）
针刀155.708.3651.167.3892.5253.105116414447037311（91.93）
针刀252.237.8347.526.8791.9352.274752104043964990（92.52）
针刀353.117.9743.146.2391.6854.404314125639578087（91.74）
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测序数据质量控制结果

, figureFileSmall=null, figureFileBig=null, tableContent=
样品Raw reads/MbRaw bases/GbClean reads/MbClean bases/GbQ30/%GC/%Total readsTotal mapped reads（%）
空白150.367.5545.696.5290.7054.764568823241659835（91.18）
空白255.448.3252.407.5693.4154.105240423648549636（92.64）
空白352.057.8147.816.8391.6652.824781088443834282（91.68）
模型152.047.8148.707.0192.8752.594869794444530255（91.44）
模型253.618.0449.517.0892.2353.764951423445501233（91.89）
模型350.967.6445.746.5490.5154.554573905442532703（92.98）
针刀155.708.3651.167.3892.5253.105116414447037311（91.93）
针刀252.237.8347.526.8791.9352.274752104043964990（92.52）
针刀353.117.9743.146.2391.6854.404314125639578087（91.74）
), ArticleFig(id=1315688517506810037, tenantId=1146029695717560320, journalId=1315121837170065538, articleId=1315688508115763274, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
基因n空白组模型组针刀组F值P值
CCL230.089 ±0.0040.479 ±0.1760.101 ±0.01714.1800.004
CCL730.053 ±0.0060.110 ±0.0150.075 ±0.01218.4320.010
IL3430.112 ±0.0130.375 ±0.0340.226 ±0.01991.3230.000
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DEGs相对表达量比较

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基因n空白组模型组针刀组F值P值
CCL230.089 ±0.0040.479 ±0.1760.101 ±0.01714.1800.004
CCL730.053 ±0.0060.110 ±0.0150.075 ±0.01218.4320.010
IL3430.112 ±0.0130.375 ±0.0340.226 ±0.01991.3230.000
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基于转录组学探讨针刀干预对膝骨关节炎兔关节软骨的作用机制
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史晓伟 , 于文静 , 王东峰 , 徐萌 , 刘大鹏 , 闫泽明 , 张治多 , 胡胜华 , 史术峰
中医学报 | 实验研究 2025,40(12): 2650-2660
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中医学报 |实验研究 2025 , 40 (12) : 2650 -2660
基于转录组学探讨针刀干预对膝骨关节炎兔关节软骨的作用机制
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史晓伟, 于文静, 王东峰, 徐萌, 刘大鹏, 闫泽明, 张治多, 胡胜华, 史术峰
作者信息
  • 北京中医药大学第三附属医院，北京 100029
通讯作者:
史术峰，男，黑龙江孙吴人，医学博士，主任医师，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。E-mail:
作者简介:

史晓伟（1986 -），男，河南济源人，医学博士，副研究员，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。

Action Mechanism of Acupotomy Intervention in Rabbit Knee Osteoarthritis Cartilage Based on Transcriptomics
Xiaowei SHI, Wenjing YU, Dongfeng WANG, Meng XU, Dapeng LIU, Zeming YAN, Zhiduo ZHANG, Shenghua HU, Shufeng SHI
Affiliations
  • The Third Affiliated Hospital to Beijing University of Chinese Medicine, Beijing China 100029
出版时间: 2025-12-10 doi: 10.16368/j.issn.1674-8999.2025.12.413
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目的：基于转录组学测序技术探讨针刀干预膝骨关节炎（knee osteoarthritis，KOA）的分子机制。方法：9只新西兰兔随机分为空白组（3只）及KOA造模组（6只），造模组采用改良Videman法固定6周以建立KOA模型，拆除固定装置后根据Lequesne评分再随机分为模型组和针刀组（每组3只），干预4周。干预结束后进行行为学评价，并对兔膝关节软骨进行转录组学测序、基因本体论（Gene Ontology，GO）和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）分析。结果：（1）干预后，与模型组相比，针刀组Lequesne评分显著降低（P＜0.01）；（2）与空白组相比，模型组共筛选出1 055条差异表达基因（differentially expressed genes，DEGs），GO分析生物过程主要涉及“炎症反应”“免疫反应”和“细胞外基质组织”等功能，KEGG Level2分析显示模型组DEGs主要参与“感染性疾病”“信号转导”“免疫系统”等通路；模型组表达上调的DEGs主要参与“细胞因子相互作用”“吞噬体”等通路，而表达下调的DEGs主要参与“癌症通路”“轴突导向”等通路。与模型组相比，针刀组共筛选出312条DEGs，GO分析结果显示，这些基因主要涉及“炎症反应”“肿瘤坏死因子反应”“semaphorin-plexin信号通路”等功能，KEGG Level2分析显示其主要参与“信号转导”“感染性疾病”及“免疫系统”等通路；针刀组表达下调的DEGs参与“细胞因子相互作用”“吞噬体”“破骨细胞分化”等通路，表达上调的DEGs参与“轴突导向”“蛋白质消化和吸收”等通路。（3）进行qRT-PCR验证，结果显示：针刀干预显著降低CCL2、CCL7、IL34 mRNA表达。结论：针刀干预KOA的分子机制主要涉及关节软骨“信号转导”“免疫-炎症”等功能以及“细胞因子受体相互作用”“吞噬体”“轴突引导”等通路。

膝骨关节炎  /  针刀  /  转录组学  /  信号转导  /  免疫炎症  /  关节软骨

Objective:To explore the molecular mechanism of acupotomy intervention in knee osteoarthritis（KOA）based on transcriptome sequencing technology.Methods:Nine New Zealand rabbits were randomly divided into a blank group（3 rabbits）and a KOA modeling group（6 rabbits）.The modeling group was subjected to a modified Videman method fixation for6 weeks to establish the KOA model.After removing the fixation device，the rabbits were further randomly divided into a model group and an acupotomy group（3 rabbits each）according to Lequesne scores and intervened for 4 weeks.After the intervention，behavioral assessments were conducted，and transcriptome sequencing，Gene Ontology（GO），and Kyoto Encyclopedia of Genes and Genomes（KEGG）analyses were performed on the rabbit knee cartilage.Results:（1）After the intervention，compared with the model group，the Lequesne scores in the acupotomy group were significantly reduced（P＜0.01）；（2）Compared with the blank group，a total of 1 055 differentially expressed genes（DEGs）were identified in the model group.GO biological process analysis mainly involved functions such as ″inflammatory response″，″immune response″，and″extracellular matrix organization″.KEGG Level 2 analysis showed that the model group was mainly enriched in pathways related to ″infectious diseases″，″signal transduction″，and″immune system″.In addition，the upregulated DEGs in the model group were enriched in pathways such as″cytokine-cytokine receptor interaction″and″phagosome″，while the downregulated DEGs were enriched in ″cancer pathways″and ″axon guidance″. Compared with the model group，312 differentially expressed genes were identified in the acupotomy group.GO analysis indicated that these genes primarily involved functions such as″inflammatory response″，″tumor necrosis factor response″，and ″semaphorin-plexin signaling pathway″.KEGG Level 2 analysis showed enrichment mainly in pathways related to ″signal transduction″，″infectious diseases″，and ″immune system″.Moreover，downregulated DEGs in the acupotomy group were enriched in ″cytokine-cytokine receptor interaction″，″phagosome″，and ″osteoclast differentiation″，while upregulated DEGs were enriched in ″axon guidance″and ″protein digestion and absorption″.（3）qRT-PCR validation showed that acupotomy intervention significantly reduced the mRNA expression of CCL2，CCL7，and IL34. Conclusion:The molecular mechanism of acupotomy intervention in KOA mainly involves functions related to joint cartilage ″signal transduction″，″immune-inflammatory response″，and pathways such as ″cytokine-cytokine receptor interaction″，″phagosome″，and ″axon guidance″.

knee osteoarthritis  /  acupotomy  /  transcriptomics  /  signal transduction  /  immune-inflammatory response  /  joint cartilage
史晓伟, 于文静, 王东峰, 徐萌, 刘大鹏, 闫泽明, 张治多, 胡胜华, 史术峰. 基于转录组学探讨针刀干预对膝骨关节炎兔关节软骨的作用机制. 中医学报, 2025 , 40 (12) : 2650 -2660 . DOI: 10.16368/j.issn.1674-8999.2025.12.413
Xiaowei SHI, Wenjing YU, Dongfeng WANG, Meng XU, Dapeng LIU, Zeming YAN, Zhiduo ZHANG, Shenghua HU, Shufeng SHI. Action Mechanism of Acupotomy Intervention in Rabbit Knee Osteoarthritis Cartilage Based on Transcriptomics[J]. Acta Chinese Medicine, 2025 , 40 (12) : 2650 -2660 . DOI: 10.16368/j.issn.1674-8999.2025.12.413
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膝骨关节炎（knee osteoarthritis，KOA）是临床常见的一种慢性骨关节退行性疾病，其典型病理表现为关节软骨退变，临床表现以膝关节疼痛和关节活动受限为主[1]。KOA患病率较高，2018年的《骨关节炎诊疗指南》显示，我国KOA的发病率约8.1%[2]，同时KOA多发于中老年人，严重影响中老年人生活质量。2023年《膝骨关节炎中西医结合诊疗指南》中建议：中医综合治疗疗效确切，可贯穿全程[3]。2015年《膝骨关节炎中医诊疗专家共识》和2023年《膝骨关节炎中西医结合诊疗指南》均推荐了针刀疗法[3-4]。针刀治疗作为一种中医疗法，在KOA的临床治疗中被广泛应用。团队前期研究证实，针刀治疗可调节KOA兔膝周肌肉的生物力学，并改善关节软骨的退变[5-6]，然而，针刀对KOA兔关节软骨在全基因组水平上的调控作用尚不明确。因此，本项目拟利用转录组学测序与生物信息学分析，探索针刀对KOA兔关节软骨基因转录谱的影响，以进一步阐明其治疗KOA的分子机制。
1　材料
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1.1　实验动物
9只6月龄的健康清洁级新西兰雄兔，体质量（2.75 ±0.25）kg，购买并饲养于北京金牧阳实验动物有限公司（动物生产许可证号：SCXK 2015-0005；动物使用许可证号：SYXK 2015 -0008），所有实验动物均单笼饲养，自由获取食物和纯净水[温度（22 ±2）℃，相对湿度（45 ±5）%]。本实验研究获得北京中医药大学动物伦理委员会批准（动物伦理号：BUCM-4 -2022010101 -1097），实验过程严格按照实验动物管理规定进行。
1.2　材料与试剂
树脂绷带（揭西县泰洲医疗有限公司，泰洲）；高分子固定绷带（苏州可耐特医疗科技有限公司，KCP04）；mirVanaTM miRNA ISOlation Kit（美国Ambion，1561）；TruSeq Stranded mRNA LTSample Prep Kit（美国Illumina，RS -122 -2101）；Agencourt AMPure XP（美国Beckman Coulter，A63881）；Qubit RNA Assay Kit（美国Life Technologies，Q32852）；Bioanalyzer 2100 RNA -6000 Nano Kit（美国Aglient，5067 -1511）；SuperScript II Reverse Transcriptase（美国Invitrogen，18064014）。
1.3　仪器
AceQ ® qPCR SYBR ® Green Master Mix（Without ROX）（南京Vazyme，Q321）；GoScriptTM逆转录酶（美国Promega，A5001）；8 Strip Real-time PCR Tubes And Caps（上海生工，F603001）；超纯水仪（美国Millipore，Qso）；冷冻离心机（美国Thermo，ST16R）；凝胶成像系统（天能公司，Tanon 2500）；紫外分光光度计（美国Thermo，NanoDrop 2000）；PCR仪（美国Bio -rad，MyCycler）；磁力架（美国Invitrogen，Magnetic stand-96）；测序仪（Illumina公司，HiSeqTM2500）。
2　方法
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2.1　KOA模型制备
应用改良的Videman法（下肢过伸位固定制动）制备兔KOA模型，详细造模方法参照团队前期研究[5-7]，简要步骤如下：（1）用医用胶带缠绕左后肢；（2）以0.5 cm厚泡沫双面胶再次缠绕；（3）使树脂绷带与双面胶紧密贴合；（4）最后用高分子固定绷带缠绕树脂绷带1圈，并以弹性胶带固定。连续制动6周。
KOA兔造模成功的判断标准是关节软骨出现损伤降解，团队前期研究已证实，制动6周后的KOA兔关节软骨即出现典型OA表现[5，7]：软骨番红O/固绿染色可见软骨层着色浅淡不均、潮线消失、组织厚度明显变薄等；Mankin病理评分达到中等损伤程度；此外，反映软骨降解的分子指标COMP与CTX-Ⅱ也显著升高。
2.2　分组及干预
将9只新西兰兔按体质量随机分为两组：空白组（3只）与造模组（6只），造模组进行KOA制备（方法同前）；造模组经6周制动成功建立KOA模型后，再随机分为模型组（3只）与针刀组（3只）。在实验期间每日给予空白组与模型组相同的抓取，但不进行针刀治疗；针刀组干预与前期研究参数一致[5-6，8]，具体如下：股内侧肌、股外侧肌腱止点，股直肌肌腱止点，股二头肌肌腱止点，鹅足腱囊以及局部压痛点、结节（压痛点确定方法：根据兔子对疼痛的躲避反应来确定，当兔子出现抖动或下肢收缩等躲避反射，则该按压点即为压痛点）进行针刀松解，刀体与皮肤切面垂直刺入，每次1～2刀，出刀并加压止血；每周2次，干预4周。
2.3　行为学评价
行为学评价根据改良的Lequesne MG评分标准[7，9]进行，包括：关节疼痛（0～3分），关节活动度（0～3分），关节肿胀程度（0～2分）和步态改变（0～3分）。于干预开始前及干预结束后，对新西兰兔进行行为学评价，由两名评价人进行单独评价，取平均值进行统计分析。
2.4　转录组学测序及生物信息学分析
干预结束后，采用空气栓塞法将实验兔处死。暴露膝关节腔，快速钳取兔股骨内侧髁软骨，液氮保存备用；每组取3只进行转录组学测序及后续生物信息学分析。
建库测序主要流程：（1）提取样品总RNA并使用DNase消化DNA后，用带有Oligo（dT）的磁珠富集真核生物mRNA；（2）加入打断试剂将mRNA打断成短片段，以打断后的mRNA为模板，用六碱基随机引物合成一链cDNA，然后配制二链式反应体系合成二链cDNA，并使用试剂盒纯化双链cDNA；（3）纯化的双链cDNA再进行末端修复、加A尾并连接测序接头，进行片段大小选择，最后进行PCR扩增；（4）构建好的文库用Agilent 2100 Bioanalyzer质检合格后，使用Illumina HiSeqTM 2500测序仪进行测序，产生125 bp或150 bp的双端数据。质检合格后，使用Illumina测序仪进行测序（该实验过程由上海欧易生物医学科技有限公司全程负责完成）。
生物信息学分析流程：（1）数据预处理和质量控制：高通量测序产生的原始数据（raw reads）首先使用Trimmomatic软件[10]进行质控并去除接头（过滤掉低质量碱基以及N碱基），最终得到高质量的clean reads；将clean reads使用hisat2[11]比对到物种的参考基因组，通过基因组比对率来评估样本的情况。（2）转录本水平定量、差异表达基因（differentially expressed genes，DEGs）筛选和功能富集及聚类分析：clean reads与参考基因组比对的结果，以二进制binary文件进行储存；FPKM[12]和count使用bowtie2和eXpress软件[13]分析得到。通过eXpress软件获取样本中转录本（protein_coding）的reads数目，使用DESeq R package的estimateSizeFactors函数[14]对数据进行标准化，并使用nbinomTest函数计算差异比较的pvalue和foldchange值。挑选出pvalue小于0.05，且差异倍数大于2的差异转录本，进行GO和KEGG富集分析，以判定差异转录本主要影响的生物学功能或者通路（该分析过程由上海欧易生物科技有限公司负责完成）。
2.5　qRT-PCR结果验证
根据测序结果，选择显著差异表达的候选基因进行qRT-PCR验证。主要实验步骤如下：软骨组织使用液氮进行低温研碎，应用TRIzol法进行总RNA抽提，并进行RNA沉淀、洗涤、再悬浮等标准操作；应用NanoDrop进行总RNA浓度测定；随后应用逆转录酶试剂盒进行cDNA合成（25 ℃5 min，42 ℃60 min，70 ℃15 min）；应用AceQ®qPCR SYBR®Green Master Mix试剂盒进行qPCR反应，总体系14 μL（qPCR SYBR ® Green Master Mix 7 μL，cDNA模板3μL，上游和下游PCR引物各0.5 μL，超纯水3μL），以GAPDH基因为内参，反应程序（第一步95 ℃ 5 min；第二步95 ℃5 s，60 ℃20 s，72 ℃30 s，循环40次；第三步65 ℃5 s，95 ℃5 s）；表1为所有引物序列（上海生工提供）。mRNA相对表达量采用2 -ΔΔCq法计算。
2.6　统计学分析
采用SPSS 20.0软件（美国IBM公司）进行统计学分析，计量资料采用均值±标准差表示（±s）。数据先进行Kolmogorov-Smirnov正态性检验及Levene's test方差齐整性检验，符合正态分布及方差齐的数据资料，两组间比较采用独立样本t检验；多组间比较采用单因素方差分析（one way ANOVA），各组间分析采用LSD中Dunnett's post hoc检验。不符合正态检验或方差不齐的数据，采用非参数检验。P＜0.05为差异有统计学意义。
3　结果
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3.1　行为学评价
干预治疗前，模型组与针刀组Lequesne MG评分相比无统计学差异（P =0.374）；干预4周后，与模型组相比，针刀组Lequesne MG评分显著降低（P=0.008，P＜0.01），见表2。
3.2　转录组测序数据质量控制结果
9个新西兰兔股骨内侧髁软骨组织共获得431.67M clean reads，其中Q30值均在90%以上，clean reads比对新西兰兔参考基因组的比例均高于91.18%；说明测序数据质量控制良好，可满足后续生信分析要求，见表3。
3.3　转录组学DEGs筛选
与空白组相比，模型组共筛选出DEGs 1 055个，其中723个上调，332个下调（图1）。与模型组相比，针刀组共筛选出DEGs 312个，其中97个上调，215个下调（图2）。
3.4　DEGs GO功能富集分析
与空白组相比，对模型组的DEGs进行GO富集分析，结果显示：（1）在生物过程中，富集最显著的三个条目分别是：44个基因参与的“炎症反应”（inflammatory response，GO:0006954）、41个基因参与的“先天免疫反应”（innate immune response，GO:0045087）、35个基因参与的“免疫反应”（immune response，GO:0006955）；（2）在细胞组成中，富集最显著的3个条目分别是：111个基因参与的“细胞外区域”（extracellular region，GO:0005576）、106个基因参与的“细胞外空间”（extracellular space，GO:0005615）、85个基因参与的“质膜的组成部分”（integral component of plasma membrane，GO:0005887）；（3）在分子功能中，富集最显著的三个条目分别是：24个基因参与的“细胞外基质结构成分”（extracellular matrix structural constituent，GO:0005201）、20个基因参与的“信号受体激活”（signaling receptor activity，GO:0038023）、12个基因参与的“胶原结合域”（collagen binding，GO:0005518），见图3。
与模型组相比，对针刀组的DEGs进行GO富集分析，结果显示：（1）在生物过程中，富集最显著的三个条目分别是：17个基因参与的“炎症反应”（inflammatory response，GO:0006954）、6个基因参与的“细胞对肿瘤坏死因子反应”（cellular response to tumor necrosis factor，GO:0071356）、5个基因参与的“semaphorin-plexin信号通路”（semaphorin-plexin signaling pathway，GO:0071526）；（2）在细胞组成中，富集最显著的三个条目分别是：47个基因参与的“细胞外区域”（extracellular region，GO:0005576）、41个基因参与的“细胞外空间”（extracellular space，GO:0005615）、19个基因参与的“细胞表面”（cell surface，GO:0009986）；（3）在分子功能中，富集最显著的3个条目分别是：10个基因参与的“细胞外基质结构成分”（extracellular matrix structural constituent，GO: 0005201）、“丝氨酸型内肽酶活性”（serine-type endopeptidase activity，GO:0004252）、“钙调素结合”（calmodulin binding，GO:0005516），见图4。
3.5　DEGs KEGG通路富集分析
3.5.1　模型组vs空白组DEGs KEGG富集分析
与空白组相比，对模型组的DEGs进行KEGG通路富集分析，结果显示：39个基因参与“细胞因子受体相互作用”（cytokine -cytokine receptor interaction，path:ocu04060）、37个基因参与“吞噬体”（phagosome，path:ocu04145）、35个基因参与“EB病毒感染”（epstein -barr virus infection，path:ocu05169）。此外，KEGG Level2水平分析，模型组DEGs参与的通路主要包括：158个基因参与“infectious diseases”、157个基因参与“signal transduction”、153个基因参与“immune system”，见图5。
进一步对模型组上调和下调的DEGs分别进行KEGG富集分析，结果发现模型组DEGs表达上调的基因主要参与以下通路：37个基因参与“cytokine -cytokine receptor interaction”、36个基因参与“phagosome”、34个基因参与“epstein -barr virus infection”；富集指数（enrichment score）排名前3的分别为：“staphylococcus aureus infection”（8.849 6）、“graft-versus -host disease”（7.497 3）、“allograft rejection”（7.285 5）；模型组DEGs表达下调的基因主要参与以下通路：10个基因参与“pathways in cancer”、8个基因参与“axon guidance”、7个基因参与“wnt signaling pathway”、7个基因参与“MAPK signaling pathway”；富集指数（enrichment score）排名前3的分别为：“glycosaminoglycan biosynthesis -heparan sulfate/heparin”（12.499 2）、“alanine，aspartate and glutamate metabolism”（8.680 0）、“nicotinate and nicotinamide metabolism”（6.249 6），见图6。
3.5.2　针刀组vs模型组DEGs KEGG富集分析
与模型组相比，对针刀组的DEGs进行KEGG通路富集分析，结果显示：14个基因参与“细胞因子受体相互作用”（cytokine -cytokine receptor interaction，path:ocu04060）、12个基因参与“吞噬体”（phagosome，path:ocu04145），另外，11个基因分别参与“补体和凝血级联反应”（complement and coagulation cascades，path:ocu04610）、“破骨细胞分化”（osteoclast differentiation，path:ocu04380）、“结核病”（tu berculosis，path:ocu05152）、“神经活性配体-受体作用”（neuroactive ligand-receptor interaction，path:ocu04080）、“PI3K-Akt信号通路”（PI3K-Akt signaling pathway，path:ocu04151）。此外，进行KEGG Level2水平分析，针刀组的DEGs富集的通路主要包括：54个基因参与“信号转导”（signal transduction）、47个基因参与“感染性疾病”（infectious diseases）、39个基因参与“免疫系统”（immune system）。见图7。
进一步对针刀组上调和下调的DEGs（与模型组相比）分别进行KEGG富集分析，结果发现针刀组DEGs表达下调的基因主要涉及以下通路：13个基因参与的“cytokine -cytokine receptor interaction”、10个基因参与的“phagosome”和“osteoclast differentiation”、9个基因参与的“complement and coagulation cascades”；富集指数（enrichment score）排名前3的分别为：“staphylococcus aureus infection”（9.823 9）、“complement and coagulation cascades”（8.150 7）、“IL-17 signaling pathway”（5.698 4）。针刀组DEGs表达上调的基因主要涉及以下通路：7个基因参与的“axon guidance”、5个基因参与的“protein digestion and absorption”、4个基因参与的“adrenergic signaling in cardiomyocytes”“cGMP -PKG signaling pathway”“calcium signaling pathway”“Focal adhesion”；富集指数（enrichment score）排名前3的分别为：“amphetamine addiction”（11.445 0）、“protein digestion and absorption”（9.967 1）、“axon guidance”（8.604 9），见图8。
3.6　候选DEGs qRT-PCR验证结果
根据测序结果，针刀干预后主要富集的通路是“cytokine-cytokine receptor interaction”和“phagosome”，因此选取差异表达显著的基因进行qRT -PCR验证，选取CCL2、CCL7和IL34共3个基因。结果显示，针刀干预后，KOA软骨CCL2（P＜0.01）CCL7（P＜0. 05）和IL34（P＜0.01）mRNA相对表达量均显著降低。见表4。
4　讨论
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西医学认为KOA是一种慢性退行性关节疾病，典型病理变化是关节软骨破坏缺失[1]，同时还存在软骨损伤之外的其他病理变化，比如滑膜炎[15]、膝周肌肉萎缩[16]等，目前认为其是一种全关节疾病（whole-organ disease）。有学者认为KOA是一种生物力学和炎症共同作用的疾病[17]，突出强调了生物力学因素和炎症因素在KOA致病过程中的关键作用。
KOA在中医学总体属于“膝痹”范畴，杨威等[18]根据KOA不同分期的表现提出KOA早、中期“筋痹”尤为重要，而中、晚期属于“膝痹”“骨痹”共存，然而我们认为“筋痹”在KOA的病理过程中贯穿始终，中、晚期“筋痹”“膝痹”“骨痹”共存。这一理念即强调了“筋”的作用，“膝者，筋之府”，筋的生理功能正常与否在KOA过程中发挥重要作用。KOA广泛存在的肌肉筋膜等病变，均属于广义“筋”的范畴。“筋”是经筋理论的继承与发展，包括现代解剖学的肌肉、韧带、筋膜、关节囊等软组织。“从筋论治”“筋骨同调”治疗KOA也被众多学者所认可[18-20]。
针刀疗法已被纳入KOA治疗指南。针刀融合了西医手术刀与中医针灸针的技术特点，治疗时发挥针刺效应和手术刀切割剥离的双重作用[21]。针刀治疗KOA的作用点在“筋”，通过“调筋”而“疏利关节”，恢复膝关节内外平衡，从而发挥“治骨”作用，即“调筋治骨”。团队前期研究证实了针刀可改善KOA兔膝周肌肉生物力学、减轻肌肉萎缩程度[6]，同时对KOA兔关节软骨具有一定保护作用（病理学变化以及软骨ECM降解减少）[5]。为了探讨针刀对KOA兔软骨的保护作用具体通过哪些功能或通路来实现，本项目进行了转录组学测序和生物信息学分析，以进一步探究针刀干预KOA软骨组织的分子机制。
本项目共筛选出1 055条模型组vs空白组DEGs，GO功能富集分析其中的细胞组分显示主要富集的是细胞外区域（extracellular region）、含胶原蛋白的细胞外基质（collagen-containing extracellular matrix）等，提示KOA兔关节软骨细胞外基质发生病变，这一结果与KOA基本病理变化相一致，Chang等[22]汇总GEO数据显示模型组富集程度最高的细胞组成部分是细胞外基质（extracellular matrix）等，与本研究结果基本一致。进一步分析KOA兔GO功能富集生物过程，发现主要涉及炎症反应（inflammatory response）、先天免疫反应（innate immune response）、免疫反应（immune response）等功能；KEGG通路分析（Level2水平）发现富集通路主要是感染性疾病（infectious diseases）、免疫系统（immune system）等。这说明在KOA软骨破坏过程中炎症反应、免疫因素等发挥重要作用。对本研究模型组高表达DEGs进行KEGG分析，发现主要富集“细胞因子受体相互作用”“吞噬体”等，该结果与其他研究基本一致。Li等[23]对KOA关节液测序后进行KEGG分析，显示高表达DEGs富集通路有“细胞因子受体相互作用”“趋化因子受体通路”等。Han等[24]进行KEGG分析发现“吞噬体”“蛋白质消化”等与OA密切相关。此外，本研究模型组低表达DEGs KEGG分析主要富集“轴突引导”“Wnt信号通路”“cAMP通路”等。Chang等[22]汇集多个GEO数据，结果显示KOA KEGG富集通路有“Wnt信号通路”“轴突引导”“HIF-1信号通路”等，与本研究基本一致。以上结果说明应用Videman法成功建立KOA兔模型，众多通路参与了KOA致病过程。
本项目首次对针刀干预KOA兔软骨进行转录组学测序，结果发现针刀组vs模型组共筛选出312条DEGs，GO分析细胞成分主要富集细胞外区域（extracellular region）、细胞外空间（extracellular space）等，提示针刀干预KOA对软骨ECM发挥作用。进一步GO分析生物过程显示，主要富集炎症反应（inflammatory response）、细胞对TNF反应（cellular response to tumor necrosis factor）等，而KEGG Level2分析发现针刀DEGs富集通路包括信号转导（signal transduction）、感染性疾病（infectious diseases）和免疫系统（immune system）。上述结果表明针刀干预KOA兔主要通过信号转导、调节免疫-炎症过程而发挥软骨保护作用。此外，针刀组表达下调DEGs KEGG分析显示，富集通路主要是“cytokine-cytokine receptor interaction”“phagosome”“osteoclast differentiation”等；而针刀组表达上调DEGs KEGG分析显示，富集通路主要是“axon guidance”“protein digestion and absorption”“focal adhesion”等。针刀干预后调控的KEGG富集通路也是上文KOA模型组富集的关键通路，说明针刀干预KOA发挥软骨保护作用主要通过逆转KOA致病关键功能（比如信号转导、免疫-炎症反应）或通路（比如细胞因子受体相互作用、吞噬体和轴突引导等）而发挥作用。
针刀在临床广泛应用于治疗KOA，研究者们对针刀干预KOA的机制进行了探索，发现相关机制主要集中于炎症、自噬等领域[25-29]。卢曼[25]证实针刀通过AMPK-mTOR-ULK1信号通路调控软骨细胞自噬。佘泽宇等[26]应用蛋白质组学发现自噬相关蛋白表达变化（APOE等）。伍闲等[28]发现针刀调控lncRNA/miRNA/NF-κB通路的抗炎机制。张良志等[29]证实针刀通过NLRP3/Caspase-1通路降低关节内炎症介质发挥治疗作用。本研究富集分析发现，针刀治疗KOA涉及的功能为信号转导、免疫-炎症反应等。此外，本研究新发现针刀富集的主要通路包括“吞噬体”（phagosome）、“破骨细胞分化”（osteoclast differentiation）、“轴突引导”（axon guidance）、“黏着斑”（focal adhesion）等，最新研究发现“phagosome”“focal adhesion”通路在KOA中高富集[24]，研究者汇集多个GEO数据发现“axon guidance”是关键通路[22，30]，而“osteoclast differentiation”可由NF-κB、MAPK等炎症通路激活[31]。因此，这些新发现的通路可能是未来探索针刀治疗KOA机制的新方向。
总之，本研究通过转录组学测序发现：针刀干预KOA的分子机制主要涉及关节软骨“信号转导”“免疫-炎症反应”等功能以及“细胞因子受体相互作用”“吞噬体”等通路。然而针刀对KOA软骨以上功能或通路的影响需要进一步深入研究和验证。

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doi: 10.16368/j.issn.1674-8999.2025.12.413
  • 接收时间：2025-06-14
  • 首发时间：2026-10-10
  • 出版时间：2025-12-10
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  • 收稿日期：2025-06-14
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    北京中医药大学第三附属医院，北京 100029

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史术峰，男，黑龙江孙吴人，医学博士，主任医师，研究方向：针灸推拿防治肌肉骨骼系统疾病研究。E-mail:
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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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