Background
Osteoarthritis (OA) is one of the most prevalent and degenerative diseases with complicated pathology including articular cartilage degradation, subchondral sclerosis and synovitis. Chondrocytes play a crucial role in maintaining cartilage integrity.
Conclusion
Baicalin protects human OA chondrocytes against IL-1β-induced apoptosis and the degradation of ECM through activating autophagy via miR-766-3p/AIFM1 axis and serves as a potential therapeutic candidate for OA treatment.
Methods
Primary chondrocytes were treated with 10 ng/mL IL-1β alone, or pre-treated with 20 μM baicalin for 5 h followed by co-treatment with 20 μM baicalin and 10 ng/mL IL-1β. CCK-8 assay was used to assess cell viability, and cell apoptosis was analyzed by both PI/FITC-Annexin V staining and quantitating apoptosis-related Bcl-2, Bax and cleaved-caspase-3 expression at both protein and mRNA level by Western blotting and qRT-PCR, respectively. Chondrocytes were transfected with miRNA-766-3p mimic and autophagy flux was examined by LC3, Beclin and p62 Western blotting and by Cyto-ID assay to quantify autophagic vacuoles.
Results
Baicalin treatment decreased the apoptosis rate and the expressions of pro-apoptotic proteins induced by IL-1β, up-regulated anti-apoptotic Bcl-2 expression, and inhibited the degradation of ECM. Baicalin increased autophagy through up-regulating the autophagy markers Beclin-1 expression and LC3 Ⅱ/LC3 Ⅰ ratio and promoting autophagic flux. Contrarily, autophagy inhibition partially alleviated the beneficial effects of baicalin on ECM synthesis and anti-apoptosis in the chondrocytes treated with L-1β. Furthermore, the differential expressional profiles of miR-766-3p and apoptosis-inducing factor mitochondria-associated 1 (AIFM1) were determined in IL-1β and IL-1β + baicalin-treated chondrocytes, and we confirmed AIFM1 was a target of miR-766-3p. MiR-766-3p overexpression suppressed apoptosis and facilitated autophagy and ECM synthesis in the chondrocytes through decreasing AIFM1. Contrarily, silencing of miR-766-3p inhibited chondrocyte autophagy and promoted apoptosis, and this effect could be reversed by AIFM1 silence.