基于转录组测序技术探讨电针对膝骨关节炎兔关节软骨表达谱的影响

Impact of EA on Expression Profile of Joint Cartilage in KOA Rabbits Based on Transcriptome Sequencing Technology

目的:基于转录组测序技术探讨电针干预对膝骨关节炎软骨表达谱的影响。方法:12只新西兰兔按体质量随机分为空白组(4只)及KOA造模组(8只),KOA造模组应用改良Videman法制动6周复制KOA兔模型,拆除固定装置后根据Lequesne评分再随机分为模型组和电针组(每组4只),干预4周。干预后进行行为学评价,对兔膝关节软骨进行转录组学测序并进行GO和KEGG富集等生物信息学分析。结果:干预后,与模型组比较,电针组Lequesne评分显著降低(P<0.01);与空白组比较,模型组共筛选出1 055条差异基因,GO分析生物过程主要涉及炎症反应、免疫反应、血管生成和细胞外基质组织等功能,KEGG分析主要富集细胞因子受体相互作用、吞噬体、EB病毒感染和细胞粘附等通路;与模型组比较,电针组初步筛选出396条差异表达基因,进一步GO分析(生物过程)主要涉及炎症反应、免疫反应以及细胞外基质组织等功能,KEGG分析主要富集细胞因子受体相互作用、吞噬体、补体和凝血级联反应等通路;通过交集分析得到电针干预KOA的主要分子机制为白细胞激活、对细菌反应和炎症反应等白细胞-炎症反应,防御反应、免疫反应和对外部刺激反应等免疫功能。结论:在关节软骨层面,电针干预KOA主要通过影响白细胞-炎症反应、免疫功能等表达谱变化而发挥治疗作用。

Objective:To explore the impact of electro-acupuncture(EA)on expression profile of joint cartilage in rabbits with knee osteoarthritis(KOA)based on transcriptome sequencing technology.Methods:12 New Zealand rabbits were randomly divided into the blank group(n=4)and the KOA modeling group(n=8).The KOA model was established by modified Videman method for 6 weeks.After removing the fixed device,rabbits were randomly divided into the model group(n=4)and the EA group(n=4)according to the Lequesne score.After 4 weeks of intervention,the behavioral evaluation was conducted and the gene expression profiles of knee cartilage were detected by transcriptome sequencing and the function analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed.Results:The Lequesne score was significantly reduced in the EA group compared with that in the model group after intervention(P<0.01).Differential expression analysis showed that 1055 differentially expressed genes were screened out in the model group compared with those in the control group.GO functional classification analysis showed that these different expression genes mainly involved in inflammatory response,immune response,angiogenesis and extracellular matrix organization.KEGG analysis mainly enriched pathways including cytokine-cytokine receptor interaction,phagosome,EB viral infection and cell adhesion molecules.Compared with the model group,the EA group screened a total of 396 different expression genes,with the GO analysis revealing that biological processes mainly involved inflammatory response,immune response and extracellular matrix organization,and KEGG analysis mainly enriching pathways including cytokine-cytokine receptor interaction,phagosome,complement and coagulation cascades.Through intersection analysis,the main mechanisms of EA intervention in KOA were the leukocyte-inflammatory response,such as leukocyte activation,response to bacterium and inflammatory response and the immune function including defense response,immune response and response to external stimulus.Conclusion:EA intervention in KOA could exert therapeutic effects,mainly through the expression profiles of leukocyte-inflammatory response and immune function at the articular cartilage level.