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. 2023 Apr 5:14:1096616.
doi: 10.3389/fgene.2023.1096616. eCollection 2023.

Clinical efficacy of Yiqi Yangxue formula on knee osteoarthritis and unraveling therapeutic mechanism through plasma metabolites in rats

Ting Zhao  1 , Shiqi Wang  1 , Wenbin Liu  2 , Jiayan Shen  1 , Youwu Dai  3 , Mingqin Shi  3 , Xiaoyi Huang  3 , Yuanyuan Wei  3 , Tao Li  4 , Xiaoyu Zhang  1 , Zhaohu Xie  3 , Na Wang  5 , Dongdong Qin  3 , Zhaofu Li  3
Affiliations

Clinical efficacy of Yiqi Yangxue formula on knee osteoarthritis and unraveling therapeutic mechanism through plasma metabolites in rats

Ting Zhao et al. Front Genet. .

Abstract

Objective: To observe the clinical efficacy and safety of Yiqi Yangxue formula (YQYXF) on knee osteoarthritis (KOA), and to explore the underlying therapeutic mechanism of YQYXF through endogenous differential metabolites and their related metabolic pathways. Methods: A total of 61 KOA patients were recruited and divided into the treatment group (YQYXF, 30 cases) and the control group (celecoxib, Cxb, 31 cases). Effects of these two drugs on joint pain, swelling, erythrocyte sedimentation rate (ESR) and c-reactive protein (CRP) were observed, and their safety and adverse reactions were investigated. In animal experiments, 63 SD rats were randomly divided into normal control (NC) group, sham operation (sham) group, model (KOA) group, Cxb group, as well as low-dose (YL), medium-dose (YM), and high-dose groups of YQYXF (YH). The KOA rat model was established using a modified Hulth method. Ultra-high-performance liquid chromatography/Q Exactive HF-X Hybrid Quadrupole-Orbitrap Mass (UHPLC-QE-MS)-based metabolomics technology was used to analyze the changes of metabolites in plasma samples of rats. Comprehensive (VIP) >1 and t-test p < 0.05 conditions were used to screen the disease biomarkers of KOA, and the underlying mechanisms of YQYXF were explored through metabolic pathway enrichment analysis. The related markers of YQYXF were further verified by ELISA (enzyme-linked immunosorbent assay). Results: YQYXF can improve joint pain, swelling, range of motion, joint function, Michel Lequesen index of severity for osteoarthritis (ISOA) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, ESR, and CRP. No apparent adverse reactions were reported. In addition, YQYXF can improve cartilage damage in KOA rats, reverse the abnormal changes of 16 different metabolites, and exert an anti-KOA effect mainly through five metabolic pathways. The levels of reactive oxygen species (ROS) and glutathione (GSH) were significantly decreased after the treatment of YQYXF. Conclusion: YQYXF can significantly improve the clinical symptoms of KOA patients without obvious adverse reactions. It mainly improved KOA through modulating lipid metabolism-related biomarkers, reducing lipid peroxidation and oxidative stress.

Keywords: YQYXF; biomarkers; clinical efficacy; knee osteoarthritis; metabolome.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Clinical efficacy assessment. (A) Disease effectiveness comparison. (B) Joint pain VAS score. (C) Joint swelling score. (D) Joint range of motion (ROM) score. (E) Michel Lequesen index of severity for osteoarthritis (ISOA) score. (F) WOMAC score. (G) Erythrocyte sedimentation rate (ESR). (H) C-reactive protein (CRP). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
FIGURE 2
FIGURE 2
The total ion chromatograms (TIC) of YQYXF that were obtained in positive ion mode and negative ion mode. (A) UHPLC-QE-MS analysis base peak intensity chromatograms of YQYXF in positive ion mode. (B) UHPLC-QE-MS analysis base peak intensity chromatograms of YQYXF in negative ion mode.
FIGURE 3
FIGURE 3
Effects of YQYXF in OA rats. (A) Time flow chart. (B) Effects of YQYXF on body weight. (C) Cartilage tissue HE staining. NC: normal control group, sham: sham operation group, OA: model group, Cxb: celecoxib, YL: low-dose groups of YQYXF, YM: medium-dose groups of YQYXF, YH: high-dose groups of YQYXF.
FIGURE 4
FIGURE 4
PCA score plots of the QC, NC, KOA, and YH groups. (A) Positive ion modes. (B) Negative ion modes. NC group (n = 5), KOA group (n = 6), YH group (n = 6).
FIGURE 5
FIGURE 5
OPLS-DA analysis of serum of mice. OPLS-DA scores plots: KOA vs. NC (A1: positive ion, A2: negative ion), YH vs. KOA (C1: positive ion, C2: negative ion). Permutation test of OPLS-DA model: KOA vs. NC (B1: positive ion, B2: negative ion); YH vs. KOA (D1: positive ion, D2: negative ion).
FIGURE 6
FIGURE 6
Multivariate statistical analysis of metabolite profiles in plasma. Volcano plot of comparison groups: KOA vs. NC (A1: positive ion, A2: negative ion); YH vs. KOA (C1: positive ion, C2: negative ion). Heatmap of comparison groups: KOA vs. NC (C1: positive ion, C2: negative ion); YH vs. KOA (D1: positive ion, D2: negative ion). Screening of differential metabolites by metabolomic analysis. Significantly upregulated metabolites are shown in red, significantly downregulated metabolites in blue, and non-significantly different metabolites in grey.
FIGURE 7
FIGURE 7
Metabolic profiling of 16 potential biomarkers. (A) Principal components analysis score plot of 16 differential metabolites. (B) Metabolic pathway bubble plot of 16 differential metabolites. (C) Heatmap of correlation analysis. The horizontal and vertical coordinates in the figure represent the contrasting differential metabolites. Red represents a positive correlation, blue represents a negative correlation, and the darker the color, the stronger the correlation. Significant correlations are marked with an asterisk (*).
FIGURE 8
FIGURE 8
The effect of YQYXF on anti-oxidative stress and lipid peroxidation in OA rats. (A) Levels of ROS in plasma. (B) Levels of GSH in plasma. All data were expressed as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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References

    1. Abbott J. H., Robertson M. C., Chapple C., Pinto D., Wright A. A., Leon de la Barra S., et al. (2013). Manual therapy, exercise therapy, or both, in addition to usual care, for osteoarthritis of the hip or knee: A randomized controlled trial. 1: Clinical effectiveness. Osteoarthr. Cartil. 21, 525–534. 10.1016/j.joca.2012.12.014 - DOI - PubMed
    1. Anil U., Markus D. H., Hurley E. T., Manjunath A. K., Alaia M. J., Campbell K. A., et al. (2021). The efficacy of intra-articular injections in the treatment of knee osteoarthritis: A network meta-analysis of randomized controlled trials. Knee 32, 173–182. 10.1016/j.knee.2021.08.008 - DOI - PubMed
    1. Bolduc J. A., Collins J. A., Loeser R. F. (2019). Reactive oxygen species, aging and articular cartilage homeostasis. Free Radic. Biol. Med. 132, 73–82. 10.1016/j.freeradbiomed.2018.08.038 - DOI - PMC - PubMed
    1. Brosseau L., Yonge K. A., Robinson V., Marchand S., Judd M., Wells G., et al. (2003). Thermotherapy for treatment of osteoarthritis. Cochrane Database Syst. Rev. 2003, Cd004522. 10.1002/14651858.Cd004522 - DOI - PMC - PubMed
    1. Carlson A. K., Rawle R. A., Wallace C. W., Brooks E. G., Adams E., Greenwood M. C., et al. (2019). Characterization of synovial fluid metabolomic phenotypes of cartilage morphological changes associated with osteoarthritis. Osteoarthr. Cartil. 27, 1174–1184. 10.1016/j.joca.2019.04.007 - DOI - PMC - PubMed