Zhang Yushen, Sun Liguo, Liu Xincheng, Zhu Dongze, Dang Jingyi, Xue Yingsen, Fan Hongbin
Department of Orthopedic Surgery, Xi-jing Hospital, Fourth Military Medical University, Xi'an, China.
Department of Orthopedic Surgery, Northwest Women and Children's Hospital, Xi'an, China.
Ann Transl Med. 2021 Feb;9(3):249. doi: 10.21037/atm-20-4023.
Osteoarthritis (OA) is a common degenerative disease with multifactorial etiology. The dedifferentiation of chondrocytes can accelerate the progress of OA. Tanshinone IIA (TIIA) has been widely used to treat OA for many years and has proved to be effective in inhibiting chondrocyte dedifferentiation. Until now, the precise mechanism of TIIA's effect against dedifferentiation has not been well understood.
The targets of TIIA were explored from public databases using various methods. The related targets of OA were obtained from the GeneCards database and the Online Mendelian Inheritance in Man (OMIM) database. The potential targets and signaling pathways were determined using protein-protein interaction (PPI), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Cell viability, proliferation, and metabolic activity were analyzed . The effects of TIIA on chondrocyte dedifferentiation were evaluated by assessing morphological changes, glycosaminoglycan (GAG) production, and messenger RNA (mRNA) levels of cartilage-related genes. After 48 hours of culture in medium with 100 μg/mL TIIA, chondrocytes/hydrogel spheres were implanted to repair cartilage defects in a rat model. The harvested specimens were examined with hematoxylin and eosin (H&E) staining and immunohistochemistry to evaluate cartilage regeneration.
The results showed that there were 28 genes potentially interacting in the TIIA-chondrocyte dedifferentiation network, and nine hub genes were identified. experiments showed an inhibitory effect of TIIA on chondrocyte dedifferentiation. The proliferation and viability of chondrocytes were promoted by TIIA at a concentration of 100-200 μg/mL, but inhibited by TIIA at 400 μg/mL. Furthermore, the histology results showed that chondrocyte/hydrogel spheres pre-treated with TIIA had better cartilage repair.
This study revealed a systematic network pharmacology approach and provided a basis for the future study of TIIA as an effective treatment for cartilage regeneration. Moreover, and results confirmed the protective effects of TIIA against chondrocyte dedifferentiation.
骨关节炎（OA）是一种病因多因素的常见退行性疾病。软骨细胞去分化会加速OA的进展。丹参酮IIA（TIIA）多年来一直被广泛用于治疗OA，并且已被证明在抑制软骨细胞去分化方面有效。迄今为止，TIIA抗去分化作用的确切机制尚未完全明确。
使用多种方法从公共数据库中探索TIIA的靶点。OA的相关靶点从GeneCards数据库和《人类孟德尔遗传在线》（OMIM）数据库中获取。使用蛋白质-蛋白质相互作用（PPI）、基因本体论（GO）和京都基因与基因组百科全书（KEGG）确定潜在靶点和信号通路。分析细胞活力、增殖和代谢活性。通过评估形态变化、糖胺聚糖（GAG）产生以及软骨相关基因的信使核糖核酸（mRNA）水平来评估TIIA对软骨细胞去分化的影响。在含有100μg/mL TIIA的培养基中培养48小时后，将软骨细胞/水凝胶球植入大鼠模型以修复软骨缺损。收获的标本用苏木精和伊红（H&E）染色及免疫组织化学检查以评估软骨再生。
结果显示，在TIIA-软骨细胞去分化网络中存在28个潜在相互作用的基因，并鉴定出9个枢纽基因。实验表明TIIA对软骨细胞去分化有抑制作用。TIIA在100 - 200μg/mL浓度下促进软骨细胞的增殖和活力，但在400μg/mL时抑制软骨细胞的增殖和活力。此外，组织学结果表明，用TIIA预处理的软骨细胞/水凝胶球具有更好的软骨修复效果。
本研究揭示了一种系统的网络药理学方法，并为未来将TIIA作为软骨再生的有效治疗方法的研究提供了依据。此外，体外和体内结果证实了TIIA对软骨细胞去分化的保护作用。
