Wu Guangwen, Huang Yunmei, Chen Wenlie, Chen Junfang, Lu Xiaodong, Liu Xiangxiang, Feng Fangfang, Huang Meiya, Lin Ruhui, Li Zuanfang, Tan Chunjiang
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.
Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.
Exp Ther Med. 2020 Jun;19(6):3641-3649. doi: 10.3892/etm.2020.8624. Epub 2020 Mar 24.
The aim of the study was to observe the effects of Tougu Xiaotong capsule (TGXTC) on the microstructure and ultrastructure of meniscus in rats with early knee osteoarthritis (KOA). A total of 27 Sprague Dawley rats were randomly divided into three groups: The normal group (non-papain-induced KOA; received saline only), the model group (papain-induced KOA; received saline only) and the TGXTC group [papain-induced KOA; received TGXTC (0.31g·kg·d)]. After 4 weeks treatment, the animals were anesthetized and the sagittal plane of the intact knees (n=6 per group) was obtained and prepared in paraffin section. Following hematoxylin and eosin staining, the degeneration of cartilage structure was evaluated via Mankin score, the microstructure of meniscus was observed and the area of calcification in meniscus was analyzed. Following toluidine blue staining, the content of proteoglycan in meniscus was analyzed. Three samples in each group were obtained and the ultrathin sections of meniscus were observed through a transmission electron microscope. The results showed that compared with the normal group, in the model group the joint space became narrow and the cartilage layer was slightly damaged and the Mankin score was 4.17±0.76, suggesting that the early KOA model was successfully established. After TGXTC treatment, the joint space stenosis and cartilage damage were improved as the Mankin score significantly decreased. Compared with the normal group, in the model group the surface of meniscal cartilage was much more uneven, the area of calcification was significantly increased and the content of proteoglycan of cartilage matrix was significantly decreased. However, following TGXTC treatment, the surface of the meniscal cartilage was much more smooth and flat, and the damage of tissue structure and the calcified area were significantly reduced, and the proteoglycan of cartilage matrix content was significantly increased. Compared with the normal group, the number of cellular processes and organelles, including the rough endoplasmic reticulum, mitochondria and Golgi apparatus of meniscal cartilage were reduced and swollen in the model group. In addition, the nuclei were deformed and heterochromatin agglutinated. The extracellular collagen fibrils became slender, disordered and sparse. Compared with the model group, the TGXTC group had more cell processes and organelles, alleviated swelling and heterochromatin agglutinating. Additionally, the collagen fibrils around the cells were thicker, larger and arranged in an orderly manner. In conclusion, TGXTC exerted its therapeutic effects on the development of KOA via reducing the destruction of the cartilage structure of the meniscus and improving the composition and function of the meniscus cartilage matrix.
本研究旨在观察透骨消痛胶囊(TGXTC)对早期膝骨关节炎(KOA)大鼠半月板微观结构和超微结构的影响。将27只Sprague Dawley大鼠随机分为三组:正常组(未用木瓜蛋白酶诱导KOA;仅给予生理盐水)、模型组(用木瓜蛋白酶诱导KOA;仅给予生理盐水)和TGXTC组[用木瓜蛋白酶诱导KOA;给予TGXTC(0.31g·kg·d)]。治疗4周后,将动物麻醉,获取完整膝关节的矢状面(每组n = 6)并制成石蜡切片。苏木精-伊红染色后,通过Mankin评分评估软骨结构退变情况,观察半月板微观结构并分析半月板钙化面积。甲苯胺蓝染色后,分析半月板中蛋白聚糖含量。每组取3个样本,通过透射电子显微镜观察半月板的超薄切片。结果显示,与正常组相比,模型组关节间隙变窄,软骨层轻度受损,Mankin评分为4.17±0.76,表明成功建立了早期KOA模型。TGXTC治疗后,关节间隙狭窄和软骨损伤得到改善,Mankin评分显著降低。与正常组相比,模型组半月板软骨表面更不平整,钙化面积显著增加,软骨基质中蛋白聚糖含量显著降低。然而,TGXTC治疗后,半月板软骨表面更光滑平整,组织结构损伤和钙化面积显著减少,软骨基质中蛋白聚糖含量显著增加。与正常组相比,模型组半月板软骨的细胞突起和细胞器数量减少,包括粗面内质网、线粒体和高尔基体肿胀。此外,细胞核变形,异染色质凝集。细胞外胶原纤维变得细长、紊乱且稀疏。与模型组相比,TGXTC组细胞突起和细胞器更多,肿胀减轻,异染色质凝集减轻。此外,细胞周围的胶原纤维更粗、更大且排列有序。总之,TGXTC通过减少半月板软骨结构破坏、改善半月板软骨基质组成和功能,对KOA的发展发挥治疗作用。