Xu Liang, Zhang Feng, Cheng Gang, Yuan Xiaoyang, Wu Yujiao, Wu Huaxun, Wang Qingtong, Chen Jingyu, Kuai Jiajie, Chang Yan, Wei Wei, Yan Shangxue
Institute of Clinical Pharmacology, Anhui Medical University, 81 Meishan Road, Hefei, 230032, China.
Key Laboratory of Anti-Inflammatory and Immune Medicine, Ministry of Education, Hefei, China.
Inflamm Res. 2023 Jan;72(1):89-105. doi: 10.1007/s00011-022-01655-2. Epub 2022 Nov 4.
To explore the role and mechanism of human adipose-derived mesenchymal stem cells (hAD-MSCs) in the treatment of osteoarthritis (OA).
OA hulth model of Sprague Dawley (SD) rats and 20 ng/ml TNF-α treated chondrocytes were used as models of OA in vivo and in vitro, respectively. hAD-MSCs were administrated in the articular cavity by injection in vivo and co-cultured with chondrocytes using transwell in vitro. Haematoxylin and eosin staining and Safranin-O/Fast green staining were performed to detect tissue destruction and histopathology. Scanning electron microscopy and transmission electron microscopy were used to observe the ultrastructure of chondrocytes. The pyroptosis signaling pathway-related proteins were detected by immunohistochemistry, immunofluorescence, qRT-PCR and Western blot. And small interference technology was used to study the mechanism in depth.
hAD-MSCs could delay the development of rat OA, improve the pathological changes of joints, inhibit the expression of NLRP3, Caspase1, GSDMD and TNFR1. In vitro, the expression of pyroptosis signal proteins in chondrocytes was significantly elevated when stimulated with TNF-α, the level of inflammatory factors such as IL-1β, IL-18 was increased, and the cell morphology was significantly destroyed. While co-cultured with hAD-MSCs, these syndromes were reversed. Knockout of TNFR1 also returned the upregulation of pyroptosis signals which caused by TNF-α.
These results demonstrated that hAD-MSCs could inhibit pyroptosis signaling pathway of chondrocytes induced by TNF-α, which have raised our understanding of the role of hAD-MSCs as promising therapy for the management of OA.
探讨人脂肪间充质干细胞（hAD-MSCs）在骨关节炎（OA）治疗中的作用及机制。
分别以Sprague Dawley（SD）大鼠OA模型和20 ng/ml TNF-α处理的软骨细胞作为OA的体内和体外模型。体内通过关节腔注射给予hAD-MSCs，体外使用Transwell与软骨细胞共培养。进行苏木精-伊红染色和番红O/固绿染色以检测组织破坏和组织病理学。使用扫描电子显微镜和透射电子显微镜观察软骨细胞的超微结构。通过免疫组织化学、免疫荧光、qRT-PCR和蛋白质免疫印迹法检测焦亡信号通路相关蛋白。并采用小干扰技术深入研究其机制。
hAD-MSCs可延缓大鼠OA的发展，改善关节病理变化，抑制NLRP3、Caspase1、GSDMD和TNFR1的表达。体外实验中，TNF-α刺激时软骨细胞中焦亡信号蛋白的表达显著升高，IL-1β、IL-18等炎症因子水平增加，细胞形态明显破坏。而与hAD-MSCs共培养时，这些症状得到逆转。敲除TNFR1也使TNF-α引起的焦亡信号上调恢复正常。
这些结果表明，hAD-MSCs可抑制TNF-α诱导的软骨细胞焦亡信号通路，这提高了我们对hAD-MSCs作为OA治疗有前景疗法的作用的认识。
