Fig. 6

TIMP4 was a target of miR-92b-5p, and TIMP4 inhibition reversed the effects of miR-92b-5p knockdown on proliferation, apoptosis, and ECM degradation in LPS-induced OA chondrocytes. A The target genes predicted by TargetScan for miR-92b-5p were intersected with the significantly down-regulated mRNAs in OA cartilage from GSE113825. B The mRNAs expression of these target genes was tested by qRT-PCR in chondrocyte with miR-92b-5p overexpression or knockdown. C RIP-Ago2 analysis indicated binding of miR-92b-5p to TIMP4. D Interaction between miR-92b-5p and TIMP4 was affirmed by dual luciferase reporter assay. E Chondrocyte transfected with miR-92b-5p inhibitor or NC inhibitor was treated with 40 ng/mL LPS for 48 h. RT-qPCR was used to detect the mRNA expression of miR-92b-5p and TIMP4. F Chondrocyte was transfected with miR-92b-5p inhibitor alone or in combination with shTIMP4, and then treated with 40 ng/mL LPS for 48 h. CCK-8 was detected the viability of chondrocyte. G Chondrocyte was transfected with miR-92b-5p inhibitor alone or in combination with shTIMP4, and then treated with 40 ng/mL LPS for 48 h. The apoptosis of chondrocyte was tested by flow cytometry. H Chondrocyte was transfected with miR-92b-5p inhibitor alone or in combination with shTIMP4, and then treated with 40 ng/mL LPS for 48 h. Western blotting was applied to assess the protein expression of MMP13, ADAMTS5, Aggrecan, and Collagen II. LPS, lipopolysaccharide; NC mim., negative control corresponding to miR-92b-5p mimics; miR mim., miR-92b-5p mimics; NC inh., negative control corresponding to miR-92b-5p inhibitor; miR inh., miR-92b-5p inhibitor; shTIMP4, TIMP4 knockdown adenovirus. N = 3. ^n.s.P > 0.05, ^*P < 0.05, ^**P < 0.01, and.^***P < 0.001