加味独活寄生合剂缓解小鼠膝骨关节炎过程中肠道菌群的参与机制  

Mechanism of intestinal flora in process of knee osteoarthritis in mice alleviated by Jiawei Duhuo Jisheng Mixture

作　　者：易南星 米倚林 许晓彤 李纳平 严可[1] 邝高艳[1] 卢敏[1] YI Nan-xing;MI Yi-lin;XU Xiao-tong;LI Na-ping;YAN Ke;KUANG Gao-yan;LU Min(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学,湖南长沙410208

出　　处：《中国药理学通报》2022年第4期625-632,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81874476);湖南省自然科学基金项目(No 2020JJ5422);湖南省卫生健康委项目(No 202104070364);湖南中医药大学中医学一流学科建设课题(No 2021ZYX32)。

摘　　要：目的本实验拟通过16S rRNA测序研究加味独活寄生合剂调控肠道菌群治疗膝骨关节炎(knee osteoarthritis,KOA)的潜在机制。方法8周龄雄性C57小鼠随机分为3组:假手术组、KOA模型组、KOA模型+加味独活寄生合剂组(给药组),6只/组。手术诱导建立KOA模型,给药组以16.25 mL·kg^(-1)灌胃/天,另两组给予生理盐水,8周后取小鼠膝关节和结肠,制备成石蜡切片后行番红O-固绿、免疫组化染色观察软骨变化,16S rRNA测序观察菌群变化。结果番红O-固绿染色发现,与模型组比较,给药组明显增加了软骨面积、降低了OARSI评分;16S rRNA结果表明,加味独活寄生合剂能调控KOA小鼠肠道菌群丰富度,改善Alpha多样性(Chao1、Simpson)和Beta多样性(PCoA、NMDS)。LefSe分析显示,3组间均存在丰度差异明显的物种,包括厚结论加味独活寄生合剂能够有效保护关节软骨,延缓KOA病程进展,其机制可能是通过调节肠道菌群结构、保护肠道屏障、减轻炎症反应而实现。Aim To investigate the potential mechanism of Jiawei Duhuo Jisheng Mixture regulating intestinal flora in the treatment of knee osteoarthritis(KOA)by 16S rRNA sequencing.Methods Eight-week-old male C57 mice were randomly divided into three groups:sham group,DMM group,and model+Jiawei Duhuo Jisheng Mixture group(Mixture group),6 mice per group.KOA model was induced by destabilization of medial meniscus surgery.16.25 mL·kg^(-1)dose mixture was given daily to the mixture group,and normal saline was given to the sham and DMM group.After eight weeks,the knee joints and colons of mice were collected,and the knee joints were prepared into paraffin sections,and the cartilage changes were observed with Safranin O-Fast Green and immunohistochemistry staining.16S rRNA sequencing of intestinal contents was performed to observe the changes of intestinal flora.Results Compared with model group,Jiawei Duhuo Jisheng Mixture could significantly reduce cartilage wear and OARSI score(P=0.0335,P=0.0295).16S rRNA sequencing showed that Jiawei Duhuo Jisheng Mixtrue could change the intestinal flora richness of KOA model mice,and improve the Alpha diversity(Chao1,Simpson)and Beta diversity(PCoA,NMDS).LefSe analysis showed that there were species with significant difference in abundance among the three groups(P=0.001),mainly including Lactobacillus,Firmicutes,Proteobacteria and other species.MetaCyc analysis indicated that Jiawei Duhuo Jisheng Mixture had effects on various metabolic pathways such as fatty acid,sugar and amino acid of intestinal flora(P<0.05).Conclusions Jiawei Duhuo Jisheng Mixtrue can effectively protect the articular cartilage and delay the progression of KOA.The mechanism may be through regulating the intestinal flora structure,protecting the intestinal barrier and reducing the inflammatory response.

关 键 词：膝骨关节炎 肠道菌群 加味独活寄生合剂 16S rRNA DMM手术 肠道-关节轴 

分 类 号：R-332[医药卫生] R282.71