中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (32): 5108-5113.doi: 10.12307/2023.573

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

淫羊藿苷对膝创伤性关节炎模型大鼠的镇痛效果及机制

丰瑞兵1,黄  勇1,2,胡  昊1,2,吴  刚1,2,段小锋1,2,李朝文1,2   

  1. 1湖北省中医院脊柱外科,湖北省武汉市  430074;2武汉体育学院健康科学学院,湖北省武汉市  430079
  • 收稿日期:2022-07-20 接受日期:2022-09-19 出版日期:2023-11-18 发布日期:2023-03-22
  • 通讯作者: 黄勇,博士,主任医师,湖北省中医院脊柱外科,湖北省武汉市 430074;武汉体育学院健康科学学院,湖北省武汉市 430079
  • 作者简介:丰瑞兵,男,1991年生,湖北省汉川市人,汉族,2021年湖北中医药大学毕业,博士,主治医师,主要从事骨关节疾病的临床与基础研究。

Analgesic effect and mechanism of icariin in a rat model of post-traumatic knee arthritis

Feng Ruibing1, Huang Yong1, 2, Hu Hao1, 2, Wu Gang1, 2, Duan Xiaofeng1, 2, Li Chaowen1, 2   

  1. 1Department of Spine Surgery, Hubei Hospital of TCM, Wuhan 430074, Hubei Province, China; 2School of Health Sciences, Wuhan Sports University, Wuhan 430079, Hubei Province, China
  • Received:2022-07-20 Accepted:2022-09-19 Online:2023-11-18 Published:2023-03-22
  • Contact: Huang Yong, MD, Chief physician, Department of Spine Surgery, Hubei Hospital of TCM, Wuhan 430074, Hubei Province, China; School of Health Sciences, Wuhan Sports University, Wuhan 430079, Hubei Province, China
  • About author:Feng Ruibing, MD, Attending physician, Department of Spine Surgery, Hubei Hospital of TCM, Wuhan 430074, Hubei Province, China

摘要:

文题释义:
淫羊藿苷:是中药淫羊藿的主要活性成分,可从箭叶淫羊藿、柔毛淫羊藿等干燥茎叶中提取,呈淡黄色针状结晶粉末,相对分子质量为676.65。
创伤性关节炎: 是骨关节炎的常见类型之一,指继发于关节创伤的骨关节炎。以关节软骨的退化、变性及关节周围骨赘形成为主要病理改变,以关节疼痛、活动障碍为主要临床表现的一种疾病。

背景:创伤性关节炎是骨关节炎最常见的形式之一,体内外研究显示,淫羊藿苷能通过多靶点、多途径、多通路对骨关节炎起到治疗作用,并且无明显不良反应,但淫羊藿苷对创伤性关节炎的镇痛效果及作用机制尚未见报道。
目的:探讨淫羊藿苷对大鼠膝创伤性关节炎的镇痛效果及机制。
方法:30只雄性SD大鼠随机分为假手术组、模型组和淫羊藿苷组,每组10只。后两组切断大鼠右膝前交叉韧带造模,假手术组仅切开右膝关节囊。造模结束后,淫羊藿苷组大鼠给予淫羊藿苷溶液100 mg/(kg·d)灌胃,假手术组与模型组同时期给予等量生理盐水灌胃,连续8周。给药结束后,采用机械刺激法与热辐射法检测机械撤退阈值与热撤退潜伏期,以此评估大鼠的疼痛阈值;应用旷场实验评估大鼠焦虑、抑郁状态;对膝关节软骨组织进行大体观察及Pelletier评分;以番红固绿染色与国际骨关节炎研究学会评分评估软骨退变程度;采用ELISA检测关节液与滑膜组织炎症因子白细胞介素1β、肿瘤坏死因子α的质量浓度;蛋白印迹法检测软骨组织单核细胞趋化蛋白1/单核细胞膜受体2配体受体轴的蛋白表达水平。
结果与结论:①与模型组相比,淫羊藿苷组的机械撤退阈值与热撤退潜伏期均显著升高(P < 0.05);②与假手术组相比,模型组大鼠运动总距离、总穿格数、中央区滞留时间均显著减少(P < 0.05),与模型组相比,淫羊藿苷组大鼠上述3个指标均显著增加(P < 0.05);③与模型组相比,淫羊藿苷组膝关节液及滑膜组织中白细胞介素1β、肿瘤坏死因子α的质量浓度均显著降低(P < 0.05);④与假手术组相比,模型组关节软骨面多表现为凹凸不平、形变,软骨组织肥大、硬化无光泽,大量软骨组织剥脱;而淫羊藿苷组较模型组软骨退变明显减轻;⑤番红固绿染色与国际骨关节炎研究学会评分结果显示,与假手术组相比,模型组膝关节软骨组织退变严重,而淫羊藿苷组较模型组软骨退变明显减轻;⑥与模型组相比,淫羊藿苷组单核细胞趋化蛋白1/单核细胞膜受体2配体受体轴的蛋白表达水平显著降低(P < 0.05);⑦提示淫羊藿苷可以显著减轻大鼠膝创伤性关节炎大鼠模型的痛觉过敏,其作用机制可能与减少膝关节内炎症因子白细胞介素1β、肿瘤坏死因子α的质量浓度及抑制单核细胞趋化蛋白1/单核细胞膜受体2配体受体轴的蛋白表达有关。

https://orcid.org/0000-0003-1388-7256(丰瑞兵)

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: 淫羊藿, 创伤性关节炎, 疼痛, 炎症因子, 单核细胞趋化蛋白1, 单核细胞膜受体2

Abstract: BACKGROUND: Post-traumatic arthritis is one of the most common forms of osteoarthritis. In vivo and in vitro studies have shown that icariin can treat osteoarthritis through multiple targets and multiple pathways with no obvious side effects. However, the analgesic effect and mechanism of icariin on post-traumatic arthritis has not been reported. 
OBJECTIVE: To investigate the analgesic effect and mechanism of icariin on post-traumatic arthritis in rats. 
METHODS: Thirty male Sprague-Dawley rats were randomly divided into sham group (n=10), model group (n=10) and icariin group (n=10). The right knee anterior cruciate ligament was cut off in the model group and icariin group, while the right knee joint capsule was cut only in the sham group. The rats in the icariin group were given intragastric administration of icariin (100 mg/kg/d), and the sham and model groups were given the same amount of normal saline at the same time point for 8 weeks. Mechanical withdrawal threshold and thermal withdrawal latency were measured by mechanical stimulation and thermal radiation to evaluate the pain threshold of rats. Open field test was used to evaluate anxiety and depression. General observation and Pelletier scoring of the knee cartilage were performed. Safranin O-Fast green staining and Osteoarthritis Research Society International score were used to evaluate the degree of cartilage degeneration. ELISA was used to detect the contents of inflammatory factors interleukin-1β and tumor necrosis factor-α in synovial fluid and synovial tissue. The protein expression of monocyte chemotactic protein 1/chemokine C-C-motif receptor 2 ligand receptor axis in cartilage tissue was detected by western blot.
RESULTS AND CONCLUSION: The mechanical withdrawal threshold and thermal withdrawal latency of the icariin group were significantly higher than those of the model group (P < 0.05). Compared with the sham group, the total exercise distance, the total number of crossing squares and the residence time in the central area of the model group were significantly decreased (P < 0.05). Compared with the model group, the above three indexes were significantly increased in the icariin group (P < 0.05). Compared with the model group, the levels of interleukin-1β and tumor necrosis factor-α in knee synovial fluid and synovial tissue were significantly decreased in the icariin group (P < 0.05). Compared with the sham operation group, the articular cartilage surface of the model group was more uneven and deformed, the cartilage tissue was hypertrophy, hardened and dull, and a large amount of cartilage tissue was exfoliated, while the cartilage degeneration in the icariin group was significantly less than that in the model group. The results of Safranin O-Fast green staining and Osteoarthritis Research Society International score showed that the degeneration of knee cartilage in the model group was more severe than that in the sham group, while that in the icariin group was significantly less than that in the model group. Compared with the model group, the protein expression level of monocyte chemotactic protein 1/chemokine C-C-motif receptor 2 ligand receptor axis was significantly lower in the icariin group (P < 0.05). The above findings indicate that icariin can significantly reduce hyperalgesia in the rat model of post-traumatic knee arthritis, and its mechanism may be related to reducing the levels of interleukin-1β and tumor necrosis factor-α in the knee joint and inhibiting the protein expression of monocyte chemotactic protein 1/chemokine C-C-motif receptor 2 ligand receptor axis.

Key words: icariin, post-traumatic arthritis, pain, inflammatory factor, monocyte chemoattractant protein 1, chemokine C-C-motif receptor 2

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