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ORIGINAL RESEARCH article

Front. Anal. Sci.
Sec. Pharmaceutical analysis
Volume 5 - 2025 | doi: 10.3389/frans.2025.1533486
This article is part of the Research Topic Thought Leaders in Analytical Science Research View all 10 articles

The chemicalome profiling of Zishen Yuzhen Pill in vivo and its promoting effect on osteogenic differentiation of MC3T3-E1 cells

Provisionally accepted
Juanjuan Cheng Juanjuan Cheng 1Xinyue Meng Xinyue Meng 2Daozheng Fang Daozheng Fang 2Yong Zhu Yong Zhu 2Zhihao Liu Zhihao Liu 2Xinyue Li Xinyue Li 2Ke Jie Ke Jie 3Shiying Huang Shiying Huang 1Huilin Li Huilin Li 1Shangbin Zhang Shangbin Zhang 1Jihang Chen Jihang Chen 2Jianping Chen Jianping Chen 1*
  • 1 The Fourth Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, China
  • 2 School of Medicine, The Chinese University of Hong Kong, Shenzhen, China
  • 3 Foshan Hospital of Traditional Chinese Medicine, Foshan, Guangdong Province, China

The final, formatted version of the article will be published soon.

    Zishen Yuzhen Pill (ZYP) is a Chinese herbal product developed by Shenzhen TCM Hospital, which have been frequently used to treat osteoporosis (OP). This study aimed to determine the major chemical components of ZYP and its prototype compounds and metabolites in rat biological samples, as well as explore the potential effect of ZYPcontaining serum in MC3T3-E1 cells. UPLC-Q/TOF-MS was used to identified the chemical components. Then, ZYP was orally administered to rat, and samples of plasma, urine, feces, bile, and tissue were collected to identify prototype compounds and metabolites. The viability of MC3T3-E1 cells was evaluated using the CCK-8 method after treatment with various concentrations (2%, 4%, and 8%) of ZYP-containing serum.Following treatment of MC3T3-E1 cells with ZYP-containing serum, the activity of alkaline phosphatase (ALP) and Alizarin red S (ARS) was measured, and the levels of Runx2, Opn, Opg and Osterix were quantified using the qPCR and Western blot analysis. And cells were collected to RNA-seq analysis. Results indicated that a total 152 compounds were identified in ZYP, including flavonoids, iridoid, lignans, triterpene saponins, etc. Furthermore, we detected a total of 70 prototype components and 99 metabolites distributed in different tissues. In addition, ZYP-containing serum observably promoted osteogenesis by increasing ALP and ARS activities, as well as up-regulating the expression of Runx2, Opn, Opg and Osterix in MC3T3-E1 cells.RNA-seq results indicated that the beneficial effects may be related to the upregulation of mitochondrial oxidative phosphorylation. This work provided further support for the traditional application of ZYP in the treatment of OP. And this study can promote the further pharmacokinetic and pharmacological evaluation of ZYP.

    Keywords: Zishen Yuzhen Pill, UPLC-QTOF/MS, chemical characterization, osteoporosis, MC3T3-E1 cells Abbreviations：ABR, Achyranthis Bidentatae Radix, ALP, alkaline phosphatase, AR, Astragali Radix, BP, biological process, CC, cellular component, CR Codonopsis Radix, DR, Drynariae Rhizoma, ECL, enhanced chemiluminescence

    Received: 24 Nov 2024; Accepted: 10 Jan 2025.

    Copyright: © 2025 Cheng, Meng, Fang, Zhu, Liu, Li, Jie, Huang, Li, Zhang, Chen and Chen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Jianping Chen, The Fourth Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.