Moderate activation of Wnt/β-catenin signaling promotes the survival of rat nucleus pulposus cells via regulating apoptosis, autophagy, and senescence
Moderate activation of Wnt/β-catenin signaling promotes the survival of rat nucleus pulposus cells via regulating apoptosis, autophagy, and senescence
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Wnt/β-连环蛋白信号的适度激活通过调节细胞凋亡、自噬和衰老促进大鼠髓核细胞的存活
DOI：
10.1002/jcb.28518
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发表时间：
2019-08-01
影响因子：
4
通讯作者：
Shao, Zengwu
中科院分区：
文献类型：
--
作者：
Li, Zhiliang;Chen, Songfeng;Shao, Zengwu
本研究旨在探讨Wnt/β-catenin信号在压迫诱导的大鼠髓核(NP)细胞凋亡、自噬和衰老中的特殊作用。最初，细胞经历了不同时期的1.0兆帕压缩。对WNT/β-catenin信号相关分子进行详细分析，并选择0、24和48h的暴露时间。然后将细胞分为对照组、Wnt/β-连环蛋白抑制物(IWP-2)组、Wnt/β-连环蛋白激活剂(LiCl)组和β-连环蛋白过表达组。在压迫0、24、48h后，用Western印迹分析和实时定量聚合酶链式反应检测细胞的凋亡、自噬和衰老，并分别用Hoechst33258、单丹西林和SA-β-Gal染色进行肉眼观察。此外，还检测了Wnt/β-catenin信号对细胞形态、活力、细胞周期、死亡率和超微结构的调节作用，以全面评价NP细胞的生存能力。结果表明，压迫可引起Wnt/β-catenin信号的时间依赖性激活。IWP-2处理降低了细胞存活率，这与自噬的下调以及凋亡和衰老的增加相对应。LiCl处理使细胞存活更加有效，同时伴随着自噬的增加和凋亡和衰老的下调；然而，与LiCl相反，β-连环蛋白的过表达加剧了压缩诱导的NP细胞死亡。总之，适度激活Wnt/β-catenin信号可通过下调细胞凋亡、衰老和上调自噬而使NP细胞更有效地存活，而过度激活Wnt/β-catenin信号则达到相反的效果。旨在调节Wnt/β-catenin信号转导的治疗策略可能是改善压迫诱导的NP细胞死亡和潜在治疗椎间盘退变的新靶点。
This study aimed to investigate the specific role of Wnt/beta-catenin signaling in compression-induced apoptosis, autophagy, and senescence in rat nucleus pulposus (NP) cells. Initially, the cells underwent various periods of exposure to 1.0 MPa compression. Wnt/beta-catenin signaling associated molecules were assessed in detail, and then 0, 24 and 48 hours exposure periods were selected. The cells were then divided into control, Wnt/beta-catenin inhibitor (IWP-2), Wnt/beta-catenin activator (LiCl), and beta-catenin overexpression groups. After 0, 24, and 48 hours of compression, apoptosis, autophagy, and senescence were evaluated by Western blot analysis and real-time polymerase chain reaction and were visually observed by Hoechst33258, monodansylcadaverine, and SA-beta-gal stainings, respectively. Additionally, the regulatory effect of Wnt/beta-catenin signaling on cell morphology, viability, cell cycle, death ratio, and ultrastructure was detected to thoroughly evaluate the survival capacity of NP cells. The results established that compression elicited a time-dependent activation of Wnt/beta-catenin signaling. The IWP-2 treatment decreased cell survival rate, which corresponded to downregulation of autophagy as well as increases in apoptosis and senescence. LiCl treatment enabled more efficient of cell survival accompanied by increased autophagy and downregulated apoptosis and senescence; however, in contrast to LiCl, overexpression of beta-catenin aggravated compression-induced NP cells death. In conclusion, moderate activation of Wnt/beta-catenin signaling enables more efficient of NP cells survival via downregulation of apoptosis, senescence, and upregulation of autophagy, and overactivation of Wnt/beta-catenin signaling achieved the opposite effect. Treatment strategies that aim to regulate Wnt/beta-catenin signaling might be a novel target for improving compression-induced NP cells death and potential treatment of intervertebral disc degeneration.