Key Laboratory of Chinese Materia Medica, Heilongjiang University of Chinese Medicine, Ministry of Education, Harbin 150040, China.
Pharmaceutical College, Harbin University of Commerce, Harbin 150086, China.
Gene. 2020 Jan 5;722:144105. doi: 10.1016/j.gene.2019.144105. Epub 2019 Sep 12.
Caulophyllum robustum Maxim (CRM) is a medicinal compound of the Northeast and is commonly used in China for the treatment of rheumatic pain and rheumatoid arthritis (RA). A preliminary study found that CRM has good anti-inflammatory, analgesic and immunosuppressive effects. However, the specific links and targets for its function remain unclear. Our study aimed to provide a mechanism for the action of Caulophyllum robustum Maxim extraction (CRME) against RA and to establish a method for studying disease treatment using Chinese medicine.
The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) method was used to detect the toxicity of CRME in L929 cells, and the concentration ranges of the blank, model, and CRME drug groups were determined. Differentially expressed long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) were identified between the three groups. Gene Ontology (GO) and pathway enrichment analyses were performed to analyze the biological functions and pathways of the differentially expressed genes. Expression of Hist1h2bj, Hist1h2ba, Zfp36, Ccl3, Cxcl2 and Egr1 in the blank, model and drug groups was detected by quantitative real-time PCR (qRT-PCR), and the role of CRME on the above factors was determined to ensure consistency with the chip data.
A total of 329 significantly upregulated genes and 141 downregulated genes were identified between the blank and model groups. A total of 218 significantly upregulated genes and 191 downregulated genes were identified between the CRME drug group and model group. CRME has a significant role in multiple pathways involved in the occurrence and development of RA. Additionally, Hist1h2bj, Hist1h2ba, Zfp36, Ccl3, Cxcl2, and Egr1 were observed in modules of the lncRNA-mRNA weighted co-expression network, consistent with the chip data.
CRME has regulatory effects on inflammatory factors, the histone family, chemokines and their ligands that are related to RA-related cytokines, the RA pathway, the TNF signaling pathway, the Toll receptor-like signaling pathway, the chemokine signaling pathways and other pathways are related to the course of RA.
莨菪（Caulophyllum robustum Maxim）是东北的一种药用化合物，在中国常用于治疗风湿痛和类风湿关节炎（RA）。初步研究发现，莨菪具有良好的抗炎、镇痛和免疫抑制作用。然而，其功能的具体环节和靶点仍不清楚。我们的研究旨在为莨菪提取物（CRME）治疗 RA 的作用提供机制，并建立一种使用中药治疗疾病的方法。
采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）法检测 CRME 在 L929 细胞中的毒性，确定空白组、模型组和 CRME 药物组的浓度范围。鉴定三组之间差异表达的长链非编码 RNA（lncRNA）和信使 RNA（mRNA）。通过基因本体论（GO）和通路富集分析，对差异表达基因的生物学功能和通路进行分析。采用定量实时 PCR（qRT-PCR）检测空白组、模型组和药物组中 Hist1h2bj、Hist1h2ba、Zfp36、Ccl3、Cxcl2 和 Egr1 的表达，确定 CRME 对上述因子的作用，确保与芯片数据一致。
空白组与模型组之间共鉴定出 329 个显著上调基因和 141 个下调基因。CRME 药物组与模型组之间共鉴定出 218 个显著上调基因和 191 个下调基因。CRME 在多个涉及 RA 发生和发展的通路中具有显著作用。此外，Hist1h2bj、Hist1h2ba、Zfp36、Ccl3、Cxcl2 和 Egr1 观察到在 lncRNA-mRNA 加权共表达网络的模块中，与芯片数据一致。
CRME 对与 RA 相关细胞因子、RA 通路、TNF 信号通路、Toll 受体样信号通路、趋化因子信号通路等通路相关的炎症因子、组蛋白家族、趋化因子及其配体具有调节作用。
