中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (20): 4205-4214.doi: 10.12307/2025.707

• 骨组织构建 bone tissue construction • 上一篇    下一篇

蒜氨酸靶向调控表皮生长因子受体和犬尿氨酸酶治疗类风湿性关节炎的分子机制

许  良，古丽米拉•木合塔尔，居博伟，李若宁   

  1. 新疆医科大学第五附属医院，新疆维吾尔自治区乌鲁木齐市  830000
  • 收稿日期:2024-07-22 接受日期:2024-08-29 出版日期:2025-07-18 发布日期:2024-12-19
  • 通讯作者: 李若宁，硕士，新疆医科大学第五附属医院，新疆维吾尔自治区乌鲁木齐市 830000
  • 作者简介:许良，女，1985年生，新疆维吾尔自治区乌鲁木齐市人，汉族，硕士，主管药师，主要从事临床药学的基础研究。
  • 基金资助:
    新疆维吾尔自治区自然科学基金项目(2022D01C580)，项目负责人：许良

Molecular mechanism of allicin-targeted regulation of epidermal growth factor receptor and kynureninase in the treatment of rheumatoid arthritis

Xu Liang, Gulimila·Muhetaer, Ju Bowei, Li Ruoning   

  1. The Fifth Affiliated Hospital of Xinjiang Medical University, Xinjiang 830000, Xinjiang Uygur Autonomous Region, China
  • Received:2024-07-22 Accepted:2024-08-29 Online:2025-07-18 Published:2024-12-19
  • Contact: Li Ruoning, MS, The Fifth Affiliated Hospital of Xinjiang Medical University, Xinjiang 830000, Xinjiang Uygur Autonomous Region, China
  • About author:Xu Liang, MS, Pharmacist-in-charge, The Fifth Affiliated Hospital of Xinjiang Medical University, Xinjiang 830000, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    the Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 2022D01C580 (to XL)

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摘要：


文题释义：
差异表达基因：该文指的是在类风湿性关节炎患者和健康对照之间表现出显著表达差异的基因。在此次研究中，通过分析GSE45291和GSE93777两个基因表达数据集，鉴定出共计6 487个差异表达基因。这些差异表达基因是通过统计学方法筛选出来的，反映了类风湿性关节炎患者中基因表达的特异性变化。进一步的网络分析显示，这些差异表达基因中的某些基因与类风湿性关节炎的病理过程密切相关，特别是在涉及细胞增殖、凋亡和免疫反应的信号通路中，提示这些基因可能是类风湿性关节炎发病机制的重要调控者。
蒜氨酸：是一种从大蒜中提取的活性化合物，具有抗炎和抗氧化作用。此次研究采用蒜氨酸处理类风湿性关节炎滑膜成纤维细胞，以探索蒜氨酸对类风湿性关节炎的潜在治疗作用，发现蒜氨酸能够显著抑制类风湿性关节炎滑膜成纤维细胞的增殖、促进其凋亡，并下调了关键基因如表皮生长因子受体和犬尿氨酸酶的表达。这些实验结果量化了蒜氨酸在分子水平上的作用机制，表明其可能通过调控细胞信号通路和改善线粒体功能来延缓类风湿性关节炎的病理进展，具有开发为治疗类风湿性关节炎药物的潜力。

背景：蒜氨酸作为大蒜中的主要活性成分具有抗炎和抗氧化作用，但蒜氨酸改善类风湿性关节炎的作用和机制仍不清楚。
目的：利用基因表达数据分析类风湿性关节炎的差异表达基因，探讨蒜氨酸在类风湿性关节炎中的调控作用。
方法：从GSE45291和GSE93777数据集中收集类风湿性关节炎的基因表达数据，进行差异表达分析。对两套数据中的共同差异表达基因进行共表达网络分析，识别与类风湿性关节炎相关性最高的模块基因，进行功能富集分析。通过SwissTargetPrediction数据库预测蒜氨酸的靶向调控基因，并与差异表达基因行交集分析，计算差异表达靶基因的受试者工作特征曲线下面积。将人永生化类风湿性关节炎滑膜成纤维细胞接种于孔板内培养24 h后分5组处理：对照组不进行任何处理，阳性药组加入甲氨蝶呤，蒜氨酸低、中、高质量浓度组分别加入20，80，160 μg/mL的蒜氨酸，处理48 h后，CCK-8法检测细胞活性，TUNEL法检测细胞凋亡，ELISA法检测氧化应激和炎症因子水平，JC-1法检测线粒体膜电位变化，RT-qPCR和Western blot检测靶基因及相关信号通路的表达。
结果与结论：①在GSE45291和GSE93777数据集中共鉴定出6 487个共同的差异表达基因，并获得了12个共表达模块，magenta模块与类风湿性关节炎的相关性最高，模块基因主要富集于磷酯酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号通路。②通过数据库预测发现7个差异表达基因被蒜氨酸靶向调控，其中表皮生长因子受体在GSE45291数据集中的受试者工作特征曲线下面积值最大，犬尿氨酸酶在GSE93777数据集中的受试者工作特征曲线下面积值最大。③与对照组比较，其他4组类风湿性关节炎滑膜成纤维细胞活性降低，细胞凋亡数量增加，活性氧、丙二醛、白细胞介素6、白细胞介素1β水平均降低，线粒体膜电位均升高，表皮生长因子受体、犬尿氨酸酶、线粒体动力相关蛋白1 mRNA与蛋白表达均降低，线粒体融合蛋白2 mRNA与蛋白表达均升高，p-AKT、p-PI3K蛋白表达均升高。④结果表明，蒜氨酸通过调控表皮生长因子受体和犬尿氨酸酶等靶基因发挥潜在的类风湿性关节炎治疗作用。
https://orcid.org/0009-0006-7260-9466(许良)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 类风湿性关节炎, 蒜氨酸, 表皮生长因子受体, 犬尿氨酸酶, 靶基因, 工程化组织构建

Abstract: BACKGROUND: Allicin, the main active ingredient in garlic, has anti-inflammatory and antioxidant effects, but the role and mechanism of allicin in ameliorating rheumatoid arthritis remain unclear.
OBJECTIVE: To analyze differentially expressed genes in rheumatoid arthritis using gene expression data and explore the regulatory role of alliin in rheumatoid arthritis.
METHODS: Gene expression data for rheumatoid arthritis were collected from the GSE45291 and GSE93777 datasets, and differential expression analysis was then performed. Co-expression network analysis was conducted on the common differentially expressed genes identified in both datasets to identify the module genes most closely related to rheumatoid arthritis, followed by functional enrichment analysis. The SwissTargetPrediction database was used to predict the target genes of allicin in the differentially expressed genes. The area under the receiver operator characteristic curve (AUC) of the differentially expressed target genes was calculated. Human immortalized rheumatoid arthritis fibroblast-like synoviocytes were seeded onto pore plates and cultured for 24 hours, and the cells were then divided into five groups. The control group was not treated; methotrexate was added to the positive drug group; and 20, 80, and 160 μg/mL 
allicin was added to the low-, medium-, and high-dose allicin groups, respectively. After 48 hours of treatment, cell activity was assessed using cell counting kit-8 assay and cell apoptosis was detected using TUNEL assay. Levels of oxidative stress and inflammatory factors were measured by ELISA. Changes in mitochondrial membrane potential were detected by JC-1 staining. The expression levels of target genes and related signaling pathways were detected by RT-qPCR and western blot.
RESULTS AND CONCLUSION: A total of 6 487 common differentially expressed genes were identified in the GSE45291 and GSE93777 datasets, and 12 co-expression modules were obtained. The magenta module had the highest correlation with rheumatoid arthritis, with module genes primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B signaling pathway. Database predictions revealed that seven differentially expressed genes were targeted by allicin, with epidermal growth factor receptor having the highest AUC value in the GSE45291 dataset and kynureninase having the highest AUC value in the GSE93777 dataset. Treatment of human immortalized rheumatoid arthritis fibroblast-like synoviocytes with allicin significantly inhibited cell activity, promoted cell apoptosis, decreased the levels of reactive oxygen species, malondialdehyde, interleukin-6, interleukin-1β, increased the expression of mitochondrial membrane potential, decreased the mRNA and protein expression of epidermal growth factor receptor, kynureninase, mitochondrial dynamin-related protein 1, elevated the mRNA and protein expression of mitochondrial fusion protein 2, and increased the protein expression of p-AKT and p-PI3K. To conclude, allicin plays the potential therapeutic effects on rheumatoid arthritis through the regulation of target genes such as epidermal growth factor receptor and kynureninase. 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: rheumatoid arthritis, allicin, epidermal growth factor receptor, kynureninase, target gene, engineered tissue construction

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