中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (17): 2670-2675.doi: 10.3969/j.issn.2095-4344.3192

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

通络止痛凝胶干预膝关节骨性关节炎模型兔的软骨炎症及退变

陈  谱1，阮安民1，周  俊1，张晓哲1，马玉峰2，宗晨钟1，王庆普2   

  1. 1北京中医药大学，北京市   100029；2北京中医药大学第三附属医院，北京市   100029
  • 收稿日期:2020-03-25 修回日期:2020-03-31 接受日期:2020-05-13 出版日期:2021-06-18 发布日期:2021-01-08
  • 通讯作者: 王庆普，主任医师，教授，博士生导师，北京中医药大学第三附属医院，北京市 100029
  • 作者简介:陈谱，男，1991年生，江西省丰城市人，汉族，北京中医药大学中医骨伤科学专业在读博士，医师，主要从事中医药防治骨与关节退变研究。
  • 基金资助:
    国家自然科学基金面上项目(81373662)，项目负责人：王庆普；北京中医药大学自主课题项目(2019-JYB-XS-232)，项目负责人：陈谱

Effect of Tongluo Analgesic Gel on cartilage inflammation and degeneration in a rabbit model of knee osteoarthritis

Chen Pu1, Ruan Anmin1, Zhou Jun1, Zhang Xiaozhe1, Ma Yufeng2, Zong Chenzhong1, Wang Qingpu2    

  1. 1Beijing University of Chinese Medicine, Beijing 100029, China; 2The Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2020-03-25 Revised:2020-03-31 Accepted:2020-05-13 Online:2021-06-18 Published:2021-01-08
  • Contact: Wang Qingpu, Chief physician, Professor, Doctoral supervisor, The Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China
  • About author:Chen Pu, MD candidate, Physician, Beijing University of Chinese Medicine, Beijing 100029, China
  • Supported by:
    the National Natural Science Foundation of China (General Project), No. 81373662 (to WQP); Independent Selected Subject Program of Beijing University of Chinese Medicine, No. 2019-JYB-XS-232 (to CP)

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摘要：

文题释义：
膝关节骨性关节炎：是一类以滑膜炎症、软骨退变并累及软骨下骨为主要病理表现的骨科常见疾病，其主要症状有膝关节疼痛、活动障碍、晨僵等，多见于老年人，尤其是女性患者。
通络止痛凝胶：通络止痛方主要含桂枝、白芍、桃仁、红花、乳香、没药、牛膝等药物，是作者导师治疗膝关节骨性关节炎的经验方。通络止痛凝胶是在通络止痛方的基础上，以卡波姆、明胶等具有良好生物相容性的化合物作为载体制备而成的，并已成功申请专利。

背景：软骨损伤是膝关节骨性关节炎的重要病理表现之一。通络止痛凝胶是在通络止痛方的基础上，以卡波姆、明胶等具有良好生物相容性的化合物作为载体制备而成的，可直接外敷于患处，更加方便患者携带与使用。
目的：观察通络止痛凝胶对于不同程度家兔膝关节骨性关节炎软骨炎症和退变的影响，初步探讨通络止痛凝胶治疗膝关节骨性关节炎的作用机制。
方法：随机将56只家兔等分为8组，即空白对照组，假手术组，轻、中、重度膝关节骨性关节炎模型组，通络止痛凝胶干预轻、中、重度膝关节骨性关节炎组。除空白对照组及假手术组外，其余各组均予以改良Hulth法建立膝关节骨性关节炎模型，轻、中、重度膝关节骨性关节炎模型组分别于造模后4，8，16周处死，通络止痛凝胶干预轻、中、重度膝关节骨性关节炎组分别在造模4，8，16周予以通络止痛凝胶外敷3周治疗，每周3次，每次1 mL。观察各组造模前后右膝关节直径变化，苏木精-伊红染色观察各组软骨形态学变化；RT-qPCR和Western blot法检测软骨组织白细胞介素1β、肿瘤坏死因子α、基质金属蛋白酶13及解聚蛋白样金属蛋白酶5的表达水平。
结果与结论：①改良Hulth法造模后的家兔右膝关节直径较空白对照组及假手术组显著增加，且随时间延长而变化(P < 0.05，P < 0.01)；予以通络止痛凝胶治疗后各组右膝关节直径均有下降，差异有显著性意义，轻、中度组变化更为显著(P < 0.01)；②苏木精-伊红染色结果显示假手术组软骨形态学变化与空白对照组无明显区别，轻、中、重度膝关节骨性关节炎模型组软骨表面凹凸不平，层次紊乱，出现分层、剥脱；予以通络止痛凝胶干预后软骨表面稍光滑、平整；③与空白对照组相比，轻、中、重度膝关节骨性关节炎模型组白细胞介素1β、肿瘤坏死因子α、基质金属蛋白酶13及解聚蛋白样金属蛋白酶5表达水平显著升高，而通络止痛凝胶可以显著抑制白细胞介素1β、肿瘤坏死因子α、基质金属蛋白酶13及解聚蛋白样金属蛋白酶5表达水平(P < 0.05，P < 0.01)，尤其是轻度和中度膝关节骨性关节炎(P < 0.01)；④结果表明，通络止痛凝胶可以显著抑制不同程度膝关节骨性关节炎模型家兔软骨炎症及延缓软骨退变，尤其是轻、中度膝关节骨性关节炎。

关键词: 骨, 关节炎, 膝关节, 软骨退变, 凝胶, 炎症, 因子, 家兔, 实验

Abstract: BACKGROUND: Cartilage injury is one of the important pathological manifestations of knee osteoarthritis (KOA). Tongluo Analgesic Gel is prepared on the basis of Tongluo Analgesic Formula, using carbomer, gelatin and other compounds with good biocompatibility as carriers, which can be directly applied to the affected area, and is more convenient for patients to carry and use.   
OBJECTIVE: To observe the effects of Tongluo Analgesic Gel on cartilage inflammation and degeneration in rabbits with different degrees of KOA, and to preliminarily explore the mechanism of Tongluo Analgesic Gel in treating KOA. 
METHODS: Fifty-six male rabbits were randomly divided into eight groups, namely: control group; sham operation group; mild KOA group; moderate KOA group; severe KOA group; Tongluo Analgesic Gel intervention for mild, moderate, and severe KOA groups. Except for the control group and the sham operation group, KOA models were established in the other groups using the modified Hulth method. Rabbits in the mild, moderate and severe KOA groups were sacrificed at 4, 8 and 16 weeks after modeling, respectively. In the three intervention groups, 3-week intervention with Tongluo Analgesic Gel were given, 1 mL once, 3 times a week beginning at 4, 8, and 16 weeks after modeling, respectively. The changes of knee joint diameter before and after modeling in each group were observed. Hematoxylin-eosin staining was performed to observe the morphological changes of cartilage tissues in each group. RT-qPCR and western blot methods were used to detect the changes of expression levels of interleukin-1β, tumor necrosis factor-α, matrix metalloproteinase-13 and disintegrin-like metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) in cartilage tissue. 
RESULTS AND CONCLUSION: The diameter of the right knee joint of rabbit models established using the modified Hulth method was significantly increased compared with the control group and sham operation group, and it changed with time (P < 0.05, P < 0.01). After treatment with Tongluo Analgesic Gel, the diameter of the right knee joint was significantly decreased, especially in the mild and moderate KOA models (P < 0.01). The hematoxylin-eosin staining results showed that the morphological changes of cartilage tissues in the sham operation group were not significantly different from those in the blank group. The cartilage surface of the mild, moderate, and severe KOA groups was uneven, with disordered layers and delamination, whereas the cartilage surface became slightly smooth and flat after intervention with Tongluo Analgesic Gel. Compared with the control group, the expression levels of interleukin-1β, tumor necrosis factor-α, matrix metalloproteinase-13 and ADAMTS-5 in the mild, moderate and severe KOA groups were significantly increased, while Tongluo Analgesic Gel could significantly inhibit these expression levels in the KOA models (P < 0.05, P < 0.01), especially in the mild and moderate KOA groups (P < 0.01). To conclude, Tongluo Analgesic Gel can significantly inhibit cartilage inflammation and delay cartilage degeneration in rabbits with different degrees of KOA, especially in mild and moderate KOA rabbits. 

Key words: bone, osteoarthritis, knee, cartilage degeneration, gel, inflammation, factor, rabbit, experiment

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