基于制备液相色谱与非靶向代谢组学筛选鹿茸促进软骨细胞增殖的关键因子

doi:10.16431/j.cnki.1671-7236.2026.01.046

Abstract

Abstract:

Objective Deer antler exhibit significant efficacy in promoting cartilage injury repair. This study aimed to identify the underlying key factor responsible for the cartilage-repairing effects of deer antler. Method Tissue from the apical growth center of fresh antlers was extracted using ultrasonic and cold-water extraction methods， respectively， with the optimal method identified by high-performance liquid chromatography （HPLC）. Preparative liquid chromatography was employed to isolate different peak fractions from the extract. Rat articular chondrocytes were isolated and cultured in vitro. The proliferative effects of the different peak fractions on these chondrocytes were compared using the CCK-8 assay. The compositional differences among the peak fractions were analyzed by untargeted metabolomics. Further validation of the effects of key substances in the extract on chondrocyte proliferation was performed using the CCK-8 assay. Result Ultrasonic extraction was identified via HPLC as an effective method for deer antler extract preparation， producing well-resolved peaks with convenient operation. Using preparative liquid chromatography， this study successfully isolated four distinct peak fractions from the extract， all exhibiting stable baselines， well-defined peaks， and minimal or no extraneous signals. Compared with other concentrations extract，0.1 mg/mL extract significantly promoted proliferation in rat articular chondrocytes （P<0.01）. Compared with other peak components， peak 1 exhibited an extremely significant promoting effect on chondrocyte proliferation （P<0.01）. Untargeted metabolomics analysis demonstrated that lipid-like metabolites comprised the majority of the extract （43.74%）， and principal component analysis （PCA） clearly separated peaks 1-4. The relative spermine content in peak 1 was extremely significantly higher than in other fractions （P<0.01）. Compared to other concentrations of spermine， 0.25 mg/mL of spermine extremely markedly enhanced chondrocyte proliferation （P<0.01）. Conclusion Antler extract significantly promotes the proliferation of rat articular chondrocytes. Preparative liquid chromatography successfully isolated four distinct peak components of the extract， and spermine in peak 1 was identified as the key factor responsible for significantly enhancing chondrocyte proliferation of deer antler.

Key words: deer antler; preparative liquid chromatography; untargeted metabolomics; chondrocyte proliferation; spermine

CLC Number:

S825

ZHAO Chen, ZHU Shuang, LI Jiping, HU Pengfei, LI Chunyi. Screening Key Factor of Deer Antler in Promoting Chondrocyte Proliferation Through Preparative Liquid Chromatography and Untargeted Metabolomics[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(1): 509-519.

Figures/Tables 9

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Screening Key Factor of Deer Antler in Promoting Chondrocyte Proliferation Through Preparative Liquid Chromatography and Untargeted Metabolomics

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