Fig. 6: AC008 accelerates OA progression through the miR-328-3p‒AQP1/ANKH axis.

a Western blotting was performed to evaluate the protein levels of AQP1 and ANKH in chondrocytes after transfection with Vector + Mimic NC, AC008 + Mimic NC, or AC008 + miR-328-3p mimic. b A CCK-8 assay was used to evaluate the viability of chondrocytes transfected with Vector + Mimic NC, AC008 + Mimic NC, or AC008 + miR-328-3p mimic. c, d The apoptosis rates (c) and the expression levels of apoptosis-associated proteins (d) were evaluated in chondrocytes after transfection with Vector+Mimic NC, AC008 + Mimic NC, or AC008 + miR-328-3p mimic. e The expression levels of ECM proteins (Aggrecan and COL2A1) and cartilage-degrading enzymes (MMP13 and ADAMTS-5) were analyzed by western blotting in chondrocytes after transfection with Vector + Mimic NC, AC008 + Mimic NC, or AC008 + miR-328-3p mimic. f A schematic diagram illustrating the experimental design for the timeline of intra-articular injection of agomir-NC, agomir-328-3p, AAV-NC, and AAV-AC008 in vivo. g Histological sections of cartilage from mice in each group were stained with safranin-O/fast green (representative images; scale bar = 100 μm). h The modified Mankin scoring system was used to evaluate articular cartilage in mice. i IHC staining was conducted to detect the expression of COL2A1 and MMP13 in cartilage samples harvested from each group (scale bar = 100 μm). MIA monosodium iodoacetate, AAV adeno-associated virus, NC negative control. The data are presented as the means ± SDs. Statistical differences were determined using two-way ANOVA (b) or unpaired two-tailed Student’s t test (c, h). ^*P < 0.05; ^**P < 0.01; ^***P < 0.001.