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郑权发,张宇,陈芳,聂晶,于莨鉴,曾光.乌头汤、四妙散对胶原诱导性关节炎DBA/1小鼠的作用研究[J].湖南中医药大学学报,2022,42(4):564-570[点击复制] |
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| 乌头汤、四妙散对胶原诱导性关节炎DBA/1小鼠的作用研究 |
| 郑权发,张宇,陈芳,聂晶,于莨鉴,曾光 |
| (湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007;中南大学化学化工学院, 湖南 长沙 410083) |
| 摘要: |
| 目的 研究中医“散寒除湿”和“清热祛湿”两种治法的代表方乌头汤和四妙散分别对胶原诱导性关节炎（collagen-induced arthritis, CIA）DBA/1小鼠的白细胞介素-17 (interleukin-17, IL-17)、转化生长因子-β (transforming growth factor β, TGF-β)、肿瘤坏死因子-α(tumor necrosis factor-α， TNF-α)表达的影响，从“以方测证”角度探讨DBA/1小鼠CIA模型的中医证候属性。方法 复制DBA/1小鼠CIA模型，将造模成功的小鼠随机分为模型组、乌头汤低剂量组、乌头汤高剂量组、四妙散低剂量组、四妙散高剂量组、甲氨蝶呤组，另设一组空白组，每组6只，连续给药35 d。观察记录关节炎指数的变化，ELISA法检测血清中IL-17、TGF-β及TNF-α的含量水平，免疫组化法检测炎症关节组织中IL-17、TGF-β及TNF-α的表达。结果 与空白组比较，模型组关节炎指数评分明显升高（P<0.01）；模型组关节滑膜增生明显，软骨表面组织变性、坏死；关节组织、血清中IL-17、TNF-α、TGF-β表达显著增高（P<0.05或P<0.01）。与模型组比较，从第21天起给药组关节炎指数评分明显下降（P<0.05或P<0.01）；关节滑膜增生、关节软骨表面损伤减轻；乌头汤低/高剂量组、四妙散高剂量组、甲氨蝶呤组关节组织中IL-17、TNF-α、TGF-β表达明显降低（P<0.05或P<0.01）；乌头汤高剂量组、四妙散低/高剂量组、甲氨蝶呤组血清中IL-17、TNF-α表达明显降低（P<0.05或P<0.01）。各给药组之间相比较：与乌头汤高剂量组比较，甲氨蝶呤组、乌头汤低剂量组、四妙散低剂量组IL-17表达明显升高（P<0.05或P<0.01）；乌头汤低剂量组TNF-α表达升高（P<0.05），其余均无统计学差异。结论 乌头汤及四妙散给药均能改善CIA小鼠模型的症状。对相关炎症因子的抑制，可能是乌头汤及四妙散的作用机制之一，推测DBA/1小鼠CIA模型或许兼有“寒痹”与“热痹”的属性。 |
| 关键词: 胶原诱导性关节炎 四妙散 乌头汤 白细胞介素-17 转化生长因子-β 肿瘤坏死因子-α 中医证候 |
| DOI：10.3969/j.issn.1674－070X.2022.04.008 |
| 投稿时间：2021-11-23 |
| 基金项目:湖南省教育厅双一流重点项目（18A209）；湖南省中医药管理局重点项目（201919）；湖南省卫生健康委2019年度科研计划课题项目（C2019093）；国家自然科学基金青年项目 （81403206）。 |
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| Effects of Wutou Decoction and Simiao Powder on collagen-induced arthritis DBA/1 mice |
| ZHENG Quanfa,ZHANG Yu,CHEN Fang,NIE Jing,YU Langjian,ZENG Guang |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;School of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan 410083, China) |
| Abstract: |
| Objective To study the effect of the two representative methods of "dispersing cold and removing dampness" and "clearing heat and removing dampness" in traditional Chinese medicine (TCM) Wutou Decoction and Simiao Powder on the expression of interleukin-17 (IL-17), transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) in DBA/1 mice with collagen-induced arthritis (CIA), and to explore the attribute of TCM syndrome of CIA model of DBA/1 mice from the angle of “measuring symptoms and signs with prescriptions”. Methods CIA model of DBA/1 mice was replicated, the successful modeling mice were randomly divided into model group, Wutou Decoction low-dose group, Wutou Decoction high-dose group, Simiao Powder low-dose group, Simiao Powder high-dose group and methotrexate group, another blank group was set up, with 6 mice in each group. After 35 consecutive days of drug administration, the changes of arthritis index were observed and recorded. The content levels of IL-17, TGF-β and TNF-α in serum were detected by ELISA, and the expression levels of IL-17, TGF-β and TNF-α in inflammatory joint tissue were detected by immunohistochemistry. Results Compared with the blank group, the arthritis index score of the model group was significantly increased (P<0.01), and there was significant synovial hyperplasia, as well as degeneration and necrosis of cartilage surface tissues. The expression levels of IL-17, TNF-α and TGF-β in the joint tissue and serum were significantly increased (P<0.05 or P<0.01). Compared with the model group, the arthritis index score of the medication administration group decreased significantly from the 21st day (P<0.05 or P<0.01); synovial hyperplasia of joints, and alleviation of surface damage of articular cartilage were observed. The expression levels of IL-17, TNF-α and TGF-β in the joint tissues of the low-dose and high-dose groups of Wutou Decoction, the Simiao Powder high-dose group and methotrexate group were significantly decreased (P<0.05 or P<0.01). The expression levels of IL-17 and TNF-α in the serum of the Wutou Decoction high-dose group and the low-dose, high-dose groups of Simiao Powder and methotrexate group were significantly decreased (P<0.05 or P<0.01). Compared between medication administration group: compared with Wutou Decoction high-dose group, the expression of IL-17 in methotrexate group, Wutou Decoction low-dose group and Simiao Powder low-dose group was significantly increased (P<0.05 or P<0.01); the expression of TNF-α was increased in Wutou Decoction low-dose group (P<0.05), and there was no significant difference in other groups. Conclusion Both Wutou Decoction and Simiao Powder can improve the symptoms of CIA mice model. The inhibition of inflammatory factors may be one of the mechanisms of action of Wutou Decoction and Simiao Powder. It is speculated that CIA model of DBA/1 mice may have both “cold arthromyodynia” and “heat arthromyodynia” properties. |
| Key words: collagen-induced arthritis Simiao Powder Wutou Decoction interleukin-17 transforming growth factor-β tumor necrosis factor-α TCM syndrom |
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