He Cailin, Wang Yang, Wen Yuqi, Li Teng, Hu En, Zeng Siqing, Yang Bo, Xiong Xingui
Institute of Integrative Medicine, Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, Changsha, 410008, China.
National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, Hunan, P.R. China, 410008.
Chin Med. 2022 Sep 16;17(1):110. doi: 10.1186/s13020-022-00670-z.
Bi Zhong Xiao decoction (BZXD), a traditional Chinese herbal formula, has been used clinically for many years to treat rheumatoid arthritis (RA). Both clinical and experimental studies have revealed that BZXD is effective in treating RA, but the mechanism remains unclear. In this study, we aimed to explore the mechanism of efficacy of BZXD through transcriptomic analysis of lncRNA and mRNA.
The combination method of ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry was used to assess the quality of BZXD. The efficacy of BZXD in treating collagen-induced arthritis (CIA) was evaluated by clinical assessment, weight changes, hematoxylin-eosin and safranin o-fast green staining, and Micro-CT. Arraystar rat lncRNA-mRNA chip technology was used to determine the lncRNA and mRNA expression profiles of the Control, CIA and BZXD groups, and to screen gene expression profiles related to the curative effect of BZXD. A lncRNA-mRNA co-expression network was constructed for the therapeutic efficacy genes. Through GO function and KEGG pathway enrichment analysis, the biological functions and signaling pathways of therapeutic efficacy genes were determined. Based on fold change and functional annotation, key differentially expressed lncRNAs and mRNAs were selected for reverse transcription-quantitative polymerase chain reaction (RT-qPCR) validation. The functions of lncRNAs targeting mRNAs were verified in vitro.
We demonstrated that BZXD could effectively reverse bone erosion. After BZXD treatment, up to 33 lncRNAs and 107 mRNAs differentially expressed genes were reversely regulated by BZXD. These differentially expressed lncRNAs are mainly involved in the biological process of the immune response and are closely related to the ECM-receptor interaction, MAPK signaling pathway, Focal adhesion, Ras signaling pathway, Antigen processing and presentation, and Chemokine signaling pathway. We identified four lncRNAs (uc.361-, ENSRNOT00000092834, ENSRNOT00000089244, ENSRNOT00000084631) and three mRNAs (Acvr2a, Cbx2, Morc4) as potential therapeutic targets for BZXD and their microarray data consistent with the RT-qPCR. In vitro experiments confirmed that silencing the lncRNAs ENSRNOT00000092834 and ENSRNOT00000084631 reversed the expression of target mRNAs.
This study elucidates the possible mechanism of BZXD reversing bone erosion in CIA rats from the perspective of lncRNA and mRNA. To provide a basis and direction for further exploration of the mechanism of BZXD in treating RA.
痹肿消汤（BZXD）是一种传统中药方剂，临床应用多年来一直用于治疗类风湿关节炎（RA）。临床和实验研究均表明BZXD对RA有效，但作用机制尚不清楚。本研究旨在通过lncRNA和mRNA的转录组分析探索BZXD的疗效机制。
采用超高效液相色谱-质谱/质谱联用方法评估BZXD的质量。通过临床评估、体重变化、苏木精-伊红和番红O-固绿染色以及Micro-CT评估BZXD治疗胶原诱导性关节炎（CIA）的疗效。采用Arraystar大鼠lncRNA-mRNA芯片技术检测对照组、CIA组和BZXD组的lncRNA和mRNA表达谱，筛选与BZXD疗效相关的基因表达谱。构建治疗疗效基因的lncRNA-mRNA共表达网络。通过GO功能和KEGG通路富集分析，确定治疗疗效基因的生物学功能和信号通路。基于倍数变化和功能注释，选择关键差异表达的lncRNA和mRNA进行逆转录-定量聚合酶链反应（RT-qPCR）验证。在体外验证靶向mRNA的lncRNA的功能。
我们证明BZXD可有效逆转骨侵蚀。BZXD治疗后，多达33个lncRNA和107个mRNA差异表达基因被BZXD反向调控。这些差异表达的lncRNA主要参与免疫反应的生物学过程，与ECM-受体相互作用、MAPK信号通路、粘着斑、Ras信号通路、抗原加工和呈递以及趋化因子信号通路密切相关。我们鉴定出4个lncRNA（uc.361-、ENSRNOT00000092834、ENSRNOT00000089244、ENSRNOT00000084631）和3个mRNA（Acvr2a、Cbx2、Morc4）作为BZXD的潜在治疗靶点，其芯片数据与RT-qPCR结果一致。体外实验证实，沉默lncRNA ENSRNOT00000092834和ENSRNOT00000084631可逆转靶mRNA的表达。
本研究从lncRNA和mRNA角度阐明了BZXD逆转CIA大鼠骨侵蚀的可能机制。为进一步探索BZXD治疗RA的机制提供依据和方向。
