2022, Vol. 26 ›› Issue (20): 3178-3183

Bushen Tiaogan Prescription containing serum in the treatment of osteoarthritis by promoting chondrocyte autophagy in rats

Fan Shuai1, Wu Chunfei1, Liang Zujian1, Xu Zhaohui2, Xie Pingjin2, Zhu Genfu1   

Abstract: BACKGROUND: Previous studies have found that Bushen Tiaogan Prescription has obvious clinical efficacy in the treatment of osteoarthritis, but there is a lack of research on its mechanism.
OBJECTIVE: To study the effect of Bushen Tiaogan Prescription containing serum on the autophagy of rat chondrocytes, and to explore its internal mechanism.
METHODS: The third-generation rat chondrocytes subcultured were randomly divided into four groups: dimethyl sulfoxide (DMSO), 10 μg/L interleukin-1β (IL-1β), 10 μg/L IL-1β+10% Bushen Tiaogan Prescription medicated serum (BSTG), and 5 μmol/L rapamycin groups, and chondrocytes in each group were treated correspondingly for 24 hours. Dansylcadaverine staining was used to detect the expression of chondrocyte autophagosomes. Real-time fluorescence quantitative PCR was used to detect the expression of autophagy-related genes ATG5 and ATG7 mRNAs, and the expression levels of Beclin1, LC3A/B, phosphorylated protein kinase B (p-AKT), and phosphorylated mammalian target of rapamycin (p-mTOR) proteins were analyzed by western blot assay.
RESULTS AND CONCLUSION: Compared with the DMSO group, the number of autophagosomes was significantly reduced in the IL-1β group. Compared with the DMSO group, the number of autophagosomes was significantly increased in the rapamycin group. Compared with the IL-1β group, the number autophagosomes was significantly increased in the 10 μg/L IL-1β+10% BSTG group. Compared with the DMSO group, the mRNA expressions of chondrocyte autophagy-related genes ATG5, ATG7 and the protein expressions of Beclin1 and LC3A/B-II/LC3A/B-I protein expression were significantly decreased in the IL-1β
group (P < 0.05, P < 0.01, P < 0.01, P < 0.05), while the protein expressions of p-AKT and p-mTOR were significantly increased (P < 0.05, P < 0.05). Compared with the DMSO group, the mRNA expressions of ATG5 and ATG7 and the protein expressions of Beclin1 and LC3A/B-II/LC3A/B-I were significantly increased in the rapamycin group (P < 0.01, P < 0.05, P < 0.05, P < 0.05), while the protein expressions of p-AKT and p-mTOR were significantly decreased (P < 0.01, P < 0.01). Compared with the IL-1β group, the mRNA expressions of ATG5 and ATG7 the protein expressions of Beclin1 and LC3A/B-II/LC3A/B-I were significantly increased in the 10 μg/L IL-1β+10% BSTG group (P < 0.01, P < 0.01, P < 0.05, P < 0.01), while the protein expressions of p-AKT and p-mTOR were significantly decreased (P < 0.05, P < 0.05). To conclude, Bushen Tiaogan Prescription containing serum increases the autophagy ability of chondrocytes by inhibiting the AKT/mTOR signaling pathway in chondrocytes and alleviating the degeneration of articular cartilage, and thereby plays a role in the treatment of osteoarthritis.
Key words: autophagy, osteoarthritis, Bushen Tiaogan Prescription, articular cartilage, AKT/mTOR