Xiao Jialing, Li Yali, Cheng Gang, Xu Guochao
Department of Stomatology, Zhejiang Hospital, Hangzhou 310013, Zhejiang, China.
Department of Dermatology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
Open Med (Wars). 2022 Apr 18;17(1):768-780. doi: 10.1515/med-2022-0463. eCollection 2022.
This study examined the potential mechanism of zoledronate on interleukin (IL)-1β-induced temporomandibular joint osteoarthritis (TMJOA) chondrocytes, using IL-1β-induced rabbit immortalized mandibular condylar chondrocytes cultured with zoledronate. Cell viability, apoptosis, mRNA, and protein expression of relevant genes involved in extracellular matrix (ECM) degradation, apoptosis, and Wnt/β-catenin signaling were examined. The involvement of the Wnt/β-catenin signaling was examined using Wnt/β-catenin inhibitor (2-(4-(trifluoromethyl)phenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-ol (XAV-939)) and activator lithium chloride (LiCl). Aggrecan and type II collagen were downregulated by zoledronate, especially with 100 nM for 48 h ( < 0.01), consistently with the upregulation of A disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) ( < 0.001), matrix metalloprotease-9 (MMP-9) ( < 0.01), caspase-3 ( < 0.001) and downregulation of proliferating cell nuclear antigen (PCNA) ( < 0.01). The apoptotic rate increased from 34.1% to 45.7% with 100 nM zoledronate for 48 h ( < 0.01). The effects of zoledronate on ADAMTs4 ( < 0.001), MMP-9 ( < 0.001), caspase-3 ( < 0.001), and PCNA ( < 0.01) were reversed by XAV-939, while LiCl increased caspase-3 expression ( < 0.01). In conclusion, zoledronate enhances IL-1β-induced ECM degradation and cell apoptosis in TMJOA chondrocytes. Wnt/β-catenin signaling might be involved in this process, but additional studies are necessary to determine the exact involvement of Wnt/β-catenin signaling in chondrocytes after zoledronate treatment.
本研究使用唑来膦酸培养白细胞介素（IL）-1β诱导的兔永生化下颌髁突软骨细胞，探讨唑来膦酸对IL-1β诱导的颞下颌关节骨关节炎（TMJOA）软骨细胞的潜在作用机制。检测细胞活力、凋亡情况以及参与细胞外基质（ECM）降解、凋亡和Wnt/β-连环蛋白信号通路的相关基因的mRNA和蛋白表达。使用Wnt/β-连环蛋白抑制剂（2-(4-(三氟甲基)苯基)-7,8-二氢-5H-硫代吡喃并[4,3-d]嘧啶-4-醇（XAV-939））和激活剂氯化锂（LiCl）检测Wnt/β-连环蛋白信号通路的参与情况。唑来膦酸可下调聚集蛋白聚糖和II型胶原蛋白，尤其是在100 nM作用48小时时（P<0.01），这与含血小板反应蛋白基序的解聚素和金属蛋白酶-4（ADAMTS-4）（P<0.001）、基质金属蛋白酶-9（MMP-9）（P<0.01）、半胱天冬酶-3（P<0.001）的上调以及增殖细胞核抗原（PCNA）的下调（P<0.01）一致。100 nM唑来膦酸作用48小时后，凋亡率从34.1%增加到45.7%（P<0.01）。XAV-939可逆转唑来膦酸对ADAMTs4（P<0.001）、MMP-9（P<0.001）、半胱天冬酶-3（P<0.001）和PCNA（P<0.01）的影响，而LiCl可增加半胱天冬酶-3的表达（P<0.01）。总之唑来膦酸可增强IL-1β诱导的TMJOA软骨细胞的ECM降解和细胞凋亡。Wnt/β-连环蛋白信号通路可能参与此过程，但需要进一步研究以确定唑来膦酸处理后Wnt/β-连环蛋白信号通路在软骨细胞中的具体作用。
