背景:椎间盘退变(IVDD)是一种以异质性为特征的多方面疾病,其中髓核(NP)细胞的细胞外基质中的分解代谢和合成代谢之间的平衡起中心作用。目前,现有的治疗方法主要集中在缓解与IVDD相关的症状,而没有针对其潜在的病理生理过程提供有效的治疗.D-甘露糖(称为甘露糖)已在各种疾病中表现出抗分解代谢特性。然而,其在IVDD中的治疗潜力尚待探索。
方法:研究开始于优化用于恢复NP细胞的甘露糖浓度。转录组学分析用于鉴定受甘露糖影响的介体,硫氧还蛋白相互作用蛋白(Txnip)基因表现出最显著的差异。随后,小干扰RNA(siRNA)技术被用来证明Txnip是甘露糖发挥其作用的关键基因。技术,如共定位分析,分子对接,和过表达测定进一步证实了甘露糖和TXNIP之间的直接调控关系。为了阐明甘露糖的作用机制,代谢组学技术被用来确定谷氨酰胺是受甘露糖影响的核心代谢产物.接下来,各种方法,包括整合的组学数据和基因表达综合(GEO)数据库,用于验证TXNIP调节谷氨酰胺的单向途径。最后,甘露糖对IVDD的治疗效果得到验证,阐明TXNIP在椎间盘内和口服处理的大鼠谷氨酰胺代谢中的机制作用。
结果:在体内和体外实验中,发现甘露糖通过抑制分解代谢在缓解IVDD方面具有有效功效。从机械的角度来看,研究表明,甘露糖通过直接靶向转录因子max样蛋白X相互作用蛋白(MondoA)发挥其抗分解代谢作用,导致TXNIP的上调。这种上调,反过来,抑制谷氨酰胺代谢,最终通过抑制丝裂原活化蛋白激酶(MAPK)途径实现其抗分解代谢作用。更重要的是,体内实验进一步证明,与椎间盘内注射相比,口服安全浓度的甘露糖可以达到有效的治疗效果。
结论:总之,通过综合的多组学分析,包括体内和体外实验,这项研究表明,甘露糖主要通过TXNIP-谷氨酰胺轴对IVDD发挥其抗分解代谢作用。这些发现提供了有力的证据支持甘露糖在临床应用中用于缓解IVDD的潜力。与现有的临床侵入性或缓解IVDD疼痛的疗法相比,甘露糖的口服给药具有更有利于临床IVDD治疗的特点。
BACKGROUND: Intervertebral disc degeneration (IVDD) is a multifaceted condition characterized by heterogeneity, wherein the balance between catabolism and anabolism in the extracellular matrix of nucleus pulposus (NP) cells plays a central role. Presently, the available treatments primarily focus on relieving symptoms associated with IVDD without offering an effective cure targeting its underlying pathophysiological processes. D-
mannose (referred to as
mannose) has demonstrated anti-catabolic properties in various diseases. Nevertheless, its therapeutic potential in IVDD has yet to be explored.
METHODS: The study began with optimizing the
mannose concentration for restoring NP cells. Transcriptomic analyses were employed to identify the mediators influenced by mannose, with the thioredoxin-interacting protein (Txnip) gene showing the most significant differences. Subsequently, small interfering RNA (siRNA) technology was used to demonstrate that Txnip is the key gene through which mannose exerts its effects. Techniques such as colocalization analysis, molecular docking, and overexpression assays further confirmed the direct regulatory relationship between mannose and TXNIP. To elucidate the mechanism of action of mannose, metabolomics techniques were employed to pinpoint glutamine as a core metabolite affected by
mannose. Next, various methods, including integrated omics data and the Gene Expression Omnibus (GEO) database, were used to validate the one-way pathway through which TXNIP regulates glutamine. Finally, the therapeutic effect of
mannose on IVDD was validated, elucidating the mechanistic role of TXNIP in glutamine metabolism in both intradiscal and orally treated rats.
RESULTS: In both in vivo and in vitro experiments, it was discovered that mannose has potent efficacy in alleviating IVDD by inhibiting catabolism. From a mechanistic standpoint, it was shown that
mannose exerts its anti-catabolic effects by directly targeting the transcription factor max-like protein X-interacting protein (MondoA), resulting in the upregulation of TXNIP. This upregulation, in turn, inhibits glutamine metabolism, ultimately accomplishing its anti-catabolic effects by suppressing the mitogen-activated protein kinase (MAPK) pathway. More importantly, in vivo experiments have further demonstrated that compared with intradiscal injections, oral administration of mannose at safe concentrations can achieve effective therapeutic outcomes.
CONCLUSIONS: In summary, through integrated multiomics analysis, including both in vivo and in vitro experiments, this study demonstrated that mannose primarily exerts its anti-catabolic effects on IVDD through the TXNIP-glutamine axis. These findings provide strong evidence supporting the potential of the use of mannose in clinical applications for alleviating IVDD. Compared to existing clinically invasive or pain-relieving therapies for IVDD, the oral administration of mannose has characteristics that are more advantageous for clinical IVDD treatment.