Huang Yansheng, Huang Yiping, Xiao Jun, Ma Yibo, Liu Yaqing, Sun Haohao, Dai Yi, Ren Qian, Wang Sibo
Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Beilin District, Xi'an, Shanxi Province 710000, China.
The First Affiliated Hospital of Shihezi University, Shihezi City, Xinjiang Uygur Autonomous Region 832000, China.
Int J Biol Macromol. 2025 Feb;288:138521. doi: 10.1016/j.ijbiomac.2024.138521. Epub 2024 Dec 12.
Osteoclast differentiation is essential for maintaining bone metabolism, and its dysregulation, particularly in the context of Echinococcus granulosus (CE) infection, can lead to severe bone loss. This study explores a novel mechanism by which CE protoscolices (PSC) drive osteoclast differentiation through the inhibition of Nrf2, followed by the upregulation of Camkk1. Transcriptome sequencing revealed a significant down-regulation of Nrf2 in cells treated with PSC. This was confirmed by Western blot and Q-PCR assays showing reduced Nrf2 protein and gene levels. In vivo studies with Nrf2 knockout mice demonstrated that the absence of Nrf2 exacerbates bone loss induced by PSC in both the spine and lower limbs, as observed through Micro-CT imaging and TRAP staining.Further investigations identified Camkk1 as a key downstream target of Nrf2. Using high-throughput sequencing and CO-IP experiments, we established that Nrf2 directly interacts with and regulates Camkk1. Functional assays indicated that PSC-induced upregulation of Camkk1 is significantly enhanced by Nrf2 knockdown, while silencing Camkk1 alone inhibits osteoclast differentiation.The therapeutic potential of this pathway was evaluated by screening small molecule inhibitors of Camkk1, with Crenolani emerging as a potent compound. In vivo administration of Crenolani in PSC-treated mice significantly alleviated bone loss in a dose-dependent manner.These findings elucidate a crucial molecular mechanism in osteoclast differentiation driven by CE infection and propose a promising therapeutic strategy for combating CE-induced bone destruction. This study advances our understanding of bone.
破骨细胞分化对于维持骨代谢至关重要,其失调,尤其是在细粒棘球绦虫(CE)感染的情况下,可导致严重的骨质流失。本研究探索了一种新机制,即CE原头节(PSC)通过抑制Nrf2,随后上调Camkk1来驱动破骨细胞分化。转录组测序显示,用PSC处理的细胞中Nrf2显著下调。蛋白质免疫印迹和定量聚合酶链反应分析证实了这一点,显示Nrf2蛋白和基因水平降低。对Nrf2基因敲除小鼠的体内研究表明,通过显微计算机断层扫描成像和抗酒石酸酸性磷酸酶染色观察到,缺乏Nrf2会加剧PSC在脊柱和下肢诱导的骨质流失。进一步研究确定Camkk1是Nrf2的关键下游靶点。通过高通量测序和免疫共沉淀实验,我们证实Nrf2直接与Camkk1相互作用并对其进行调控。功能分析表明,Nrf2敲低显著增强了PSC诱导的Camkk1上调,而单独沉默Camkk1则抑制破骨细胞分化。通过筛选Camkk1的小分子抑制剂评估了该途径的治疗潜力,克雷诺拉尼成为一种有效的化合物。在PSC处理的小鼠体内给予克雷诺拉尼以剂量依赖性方式显著减轻了骨质流失。这些发现阐明了CE感染驱动的破骨细胞分化中的关键分子机制,并提出了一种有前景的治疗策略来对抗CE诱导的骨破坏。这项研究增进了我们对骨骼的理解。