Ren Mengxin, Wang Yajing, Yuan Yue, Du Hailing, Liang Qinghua, Qin Feng, Xiong Zhili
School of Pharmacy, Shenyang Pharmaceutical University, No.26 Huatuo Rd, High & New Tech Development Zone, 117004 Benxi, Liaoning Province, PR China.
School of Pharmacy, Shenyang Pharmaceutical University, No.26 Huatuo Rd, High & New Tech Development Zone, 117004 Benxi, Liaoning Province, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Apr 15;1256:124540. doi: 10.1016/j.jchromb.2025.124540. Epub 2025 Feb 26.
According to the theory of traditional Chinese medicine, the kidney is regarded as governing the bones and dominating the storage of essence. Gushudan (GSD) is a traditional Chinese medicine prescription that has the effects of strengthening bone and nourishing the kidney. However, the mechanism of action of GSD in preventing postmenopausal osteoporosis (PMOP) rats based on the 'kidney-bone' axis remains to be further systematically investigated. In this study, an integrated kidney metabolomics method based on three MS detection modes of UHPLC-HRMS, UHPLC-MS/MS and AFADESI-MSI was developed to reveal the protective mechanism of GSD in PMOP rats. Firstly, the non-targeted metabolomics was investigated to comprehensively explore the metabolic changes in the kidneys of PMOP rats based on the UHPLC-Q-Orbitrap HRMS. Subsequently, UHPLC-MS/MS targeted metabolomics and Mass Spectrometry Imaging (MSI) techniques were combined to elucidate the preventive mechanism of GSD on PMOP through branched-chain amino acid (BCAA) metabolism. The results of the non-targeted metabolomics demonstrated that GSD significantly modulated the levels of 67 potential biomarkers, including leucine and valine, which are primarily involved in amino acid metabolism. Specifically, BCAA metabolism is notably enriched in amino acid metabolism. Compared to the control group, it was found that the levels of BCAAs were decreased and α-branched-chain keto acids (BCKAs) were increased in the model groups in the targeted metabolomics study. Moreover, MSI results showed that the changes in BCAAs content were mainly concentrated in the renal cortex. This finding confirmed the metabolic disorders of BCAA in the renal cortex of PMOP rats, and that GSD had a significant regulatory effect on this disorder. In conclusion, this study integrated three mass spectrometry techniques that validate and complement each other to revealed the anti-osteoporostic mechanism of GSD in PMOP rats and to elucidate the modern scientific connotation of the 'kidney-bone' axis based on the BCAA metabolism.
根据中医理论，肾主骨生髓。骨疏丹（GSD）是一种具有强骨补肾作用的中药方剂。然而，基于“肾-骨”轴，GSD预防绝经后骨质疏松症（PMOP）大鼠的作用机制仍有待进一步系统研究。本研究建立了一种基于超高效液相色谱-高分辨质谱（UHPLC-HRMS）、超高效液相色谱-串联质谱（UHPLC-MS/MS）和大气压辅助解吸电喷雾离子化质谱成像（AFADESI-MSI）三种质谱检测模式的肾脏整合代谢组学方法，以揭示GSD对PMOP大鼠的保护机制。首先，基于UHPLC-Q-Orbitrap HRMS对非靶向代谢组学进行研究，全面探索PMOP大鼠肾脏中的代谢变化。随后，将UHPLC-MS/MS靶向代谢组学和质谱成像（MSI）技术相结合，通过支链氨基酸（BCAA）代谢阐明GSD对PMOP的预防机制。非靶向代谢组学结果表明，GSD显著调节了67种潜在生物标志物的水平，包括主要参与氨基酸代谢的亮氨酸和缬氨酸。具体而言，BCAA代谢在氨基酸代谢中显著富集。在靶向代谢组学研究中发现，与对照组相比，模型组中BCAAs水平降低，α-支链酮酸（BCKAs）水平升高。此外，MSI结果显示，BCAAs含量的变化主要集中在肾皮质。这一发现证实了PMOP大鼠肾皮质中BCAA的代谢紊乱，以及GSD对这种紊乱具有显著的调节作用。总之，本研究整合了三种相互验证和补充的质谱技术，揭示了GSD对PMOP大鼠的抗骨质疏松机制，并基于BCAA代谢阐明了“肾-骨”轴的现代科学内涵。
