中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (8): 1180-1186.doi: 10.12307/2022.221

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

苗药熏蒸治疗膝骨关节炎模型兔可延缓细胞外基质的破坏

王宝娟1，郑曙光1，张  琪2，李田洋2   

  1. 1贵州中医药大学第一附属医院，贵州省贵阳市   550000；2贵州中医药大学，贵州省贵阳市   550000
  • 收稿日期:2021-01-06 修回日期:2021-01-08 接受日期:2021-02-18 出版日期:2022-03-18 发布日期:2021-11-02
  • 通讯作者: 郑曙光，主任医师/教授，硕士研究生导师，贵州中医药大学第一附属医院，贵州省贵阳市 550000
  • 作者简介:王宝娟，女，1992年生，河北省迁安市人，汉族，2019年贵州中医药大学毕业，硕士，主要从事中医基础理论及民族医药研究。
  • 基金资助:
    国家自然科学基金项目(81660832，81960914)，项目负责人：郑曙光

Miao medicine fumigation can delay extracellular matrix destruction in a rabbit model of knee osteoarthritis

Wang Baojuan1, Zheng Shuguang1, Zhang Qi2, Li Tianyang2   

  1. 1The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China; 2Guizhou University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China
  • Received:2021-01-06 Revised:2021-01-08 Accepted:2021-02-18 Online:2022-03-18 Published:2021-11-02
  • Contact: Zheng Shuguang, Chief physician, Professor, Master’s supervisor, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou Province, China
  • About author:Wang Baojuan, Master, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81660832 and 81960914 (both to ZSG)

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摘要：

文题释义：
苗药熏蒸疗法：是苗医常用的一种外治方法，是以中医学基本理论为指导，选用苗族民间常用中草药煮沸后产生的气雾对人体局部进行熏蒸，从而借助药物的药力及煮沸后的热力直接作用于人体疾病部位，发挥扩张局部血管、促进血液循环、温通经脉、消肿止痛等作用。
细胞外基质：细胞外基质主要由5类物质组成，即胶原蛋白、非胶原蛋白、弹性蛋白、蛋白聚糖与氨基聚糖，其在上皮或内皮细胞的基底部者为基底膜，而在细胞间黏附结构者为间质结缔组织。研究表明，细胞外基质含有大量信号分子，积极参与控制细胞的生长、极性、形状、迁移和代谢活动。
背景：前期研究显示苗药熏蒸疗法对治疗骨性关节炎有确切疗效，但其作用机制不清楚。由于滑膜组织及软骨下骨中血管过度新生是骨性关节炎发生发展的重要原因，血管内皮生长因子是体内重要的促血管新生的因子，其过度表达与基质细胞衍生因子1/CXCR4信号通路存在密切联系。
目的：以苗医熏蒸疗法作为干预因素，探讨基质细胞衍生因子1/CXCR4信号通路在骨性关节炎发生发展中的作用及苗药熏蒸疗法的治疗作用机制。
方法：健康新西兰大白兔50只，随机分为正常组、模型组、阳性对照组(扶他林软膏外用)、早期干预组、苗药熏蒸组5组，后4组采用膝关节腔注射碘乙酸钠方法建立兔膝骨性关节炎模型，2周后造模成功开始干预治疗，连续治疗6周后取材，苏木精-伊红染色观察滑膜、软骨病理变化；免疫组化方法检测滑膜组织白细胞介素1β、肿瘤坏死因子α水平；Elisa法检测关节液中基质细胞衍生因子1及血清中血管内皮生长因子水平；实时荧光定量PCR检测关节软骨组织中血管内皮生长因子mRNA表达；Western blot检测软骨组织中Ⅱ型胶原蛋白、聚集蛋白聚糖蛋白表达。
结果与结论：①与模型组比较：苗药熏蒸组滑膜及软骨组织的炎症表达被显著抑制，病理改变减轻；苗药熏蒸组滑膜白细胞介素1β及肿瘤坏死因子α的积分吸光度值、血清中血管内皮生长因子及关节液中基质细胞衍生因子1水平均降低(P < 0.05)；苗药熏蒸组软骨血管内皮生长因子mRNA表达降低(P < 0.05)；软骨中Ⅱ型胶原蛋白、聚集蛋白聚糖表达均升高(P < 0.05)；②苗药熏蒸组上述指标与阳性对照组及早期干预组比较无显著差异(P > 0.05)。实验结果表明：苗药熏蒸疗法可能是通过减少基质细胞衍生因子1与其受体CXCR4结合，抑制白细胞介素1β、肿瘤坏死因子α、血管内皮生长因子表达，延缓细胞外基质破坏，起到治疗骨性关节炎作用。

https://orcid.org/0000-0002-1833-8558 (王宝娟) 

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: 骨性关节炎, 苗药熏蒸, 血管内皮生长因子, SDF-1/CXCR4信号通路, 基质细胞衍生因子1, 兔, 动物模型

Abstract: BACKGROUND: Previous studies have shown that Miao medicine fumigation therapy has a definite effect on the treatment of osteoarthritis; however, its mechanism is unclear. Because excessive angiogenesis in synovial tissue and subchondral bone tissue is an important cause of the occurrence and development of osteoarthritis, vascular endothelial growth factor is an important pro-angiogenesis factor in the body, and the stromal cell-derived factor 1/CXCR4 signaling pathway is closely related to the overexpression of vascular endothelial growth factor. 
OBJECTIVE: To explore the role of stromal cell-derived factor 1/CXCR4 signaling pathway in the osteoarthritic occurrence and development and the therapeutic mechanism of Miao medicine fumigation on knee osteoarthritis.
METHODS: Fifty healthy New Zealand white rabbit were randomized into normal, model, positive control (voltaren ointment for external use), early intervention, and Miao medicine fumigation groups. The knee osteoarthritis model was established in the latter four groups by injecting sodium iodoacetate into the knee joint cavity of rabbits. Hematoxylin-eosin staining was used to observe the pathological changes of synovium and cartilage. Immunohistochemical method was used to detect the levels of interleukin-1β and tumor necrosis factor-α in synovial tissue. ELISA method was used to detect the level of stromal cell-derived factor 1 in synovial fluid and the level of vascular endothelial growth factor in serum. Real-time fluorescence quantitative PCR was used to detect vascular endothelial growth factor mRNA expression in articular cartilage tissue. Western blot assay was used to detect the expression of type II collagen and aggrecan protein in cartilage tissue.
RESULTS AND CONCLUSION: Compared with the model group, Miao medicine fumigation treatment significantly inhibited the inflammation in synovium and cartilage tissue and reduced its pathological changes. Compared with the model group, the integral absorbance values of interleukin-1β and tumor necrosis factor-α in synovium and the levels of vascular endothelial growth factor in serum and stromal cell-derived factor in joint fluid were significantly reduced in the Miao medicine fumigation group (P < 0.05). Compared with the model group, the serum vascular endothelial growth factor mRNA was reduced in the Miao medicine fumigation group, while type II collagen and aggrecan expressions were increased (P < 0.05). The above experimental results indicated no significant differences among Miao medicine fumigation group, positive control group and early intervention group (P > 0.05). To conclude, the Miao medicine fumigation therapy maybe by reducing the binding of stromal cell-derived factor 1 to its receptor CXCR4 inhibits the expression of interleukin-1β, tumor necrosis factor-α, and vascular endothelial growth factor, delays the destruction of extracellular matrix, and plays a therapeutic role.

Key words: osteoarthritis, Miao medicine fumigation therapy, vascular endothelial growth factor, stromal cell-derived factor 1/CXCR4 signaling pathway, stromal cell-derived factor 1, rabbit, animal model

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