Research Paper Volume 13, Issue 5 pp 7676—7690

Downregulation of microRNA-29b by DNMT3B decelerates chondrocyte apoptosis and the progression of osteoarthritis via PTHLH/CDK4/RUNX2 axis

DNMT3B inhibits OA progression in the mouse model of OA. A mouse model of OA was induced by DMM and treated with lentivirus vector harboring oe-DNMT3B and/or oe-RUNX2. (A) Co-localization of DNMT3B and collagen II in cartilage tissues of mice detected by immunofluorescence (× 400, scale bar = 25 μm). (B) The miR-29b expression and the mRNA expression of DNMT3B and RUNX2 in cartilage tissues of mice determined by RT-qPCR. (C) Histological analyses including Safranin O staining and HE staining and microscopic observation of cartilage destruction at 8 weeks post-operation (lesions of articular cartilage were indicated by black arrows, × 400, scale bar = 25 μm). (D) OARIS scores of articular cartilage. (E) Representative images (× 400, scale bar = 25 μm) and quantitation of TUNEL positive apoptotic cells in the cartilage tissues. (F) The protein expression of CDK4, Ki67, MMP3, and MMP13 in cartilage tissues normalized to β-actin measured by Western blot analysis. *p

Figure 7. DNMT3B inhibits OA progression in the mouse model of OA. A mouse model of OA was induced by DMM and treated with lentivirus vector harboring oe-DNMT3B and/or oe-RUNX2. (A) Co-localization of DNMT3B and collagen II in cartilage tissues of mice detected by immunofluorescence (× 400, scale bar = 25 μm). (B) The miR-29b expression and the mRNA expression of DNMT3B and RUNX2 in cartilage tissues of mice determined by RT-qPCR. (C) Histological analyses including Safranin O staining and HE staining and microscopic observation of cartilage destruction at 8 weeks post-operation (lesions of articular cartilage were indicated by black arrows, × 400, scale bar = 25 μm). (D) OARIS scores of articular cartilage. (E) Representative images (× 400, scale bar = 25 μm) and quantitation of TUNEL positive apoptotic cells in the cartilage tissues. (F) The protein expression of CDK4, Ki67, MMP3, and MMP13 in cartilage tissues normalized to β-actin measured by Western blot analysis. *p < 0.05. Statistical data were measurement data and described as the mean ± standard deviation. The one-way ANOVA was used for comparison among multiple groups, followed by Tukey’s post hoc test. The experiment was repeated 3 times independently.