Melatonin Prevents Osteoarthritis-Induced Cartilage Degradation via Targeting MicroRNA-140

Figure 6

Inhibition of miR-140 intensified IL-1β-induced matrix degradation in human chondrocytes. Chondrocytes were treated with 5 ng/mL IL-1β and 100 μM melatonin (MT). To inhibit miR-140 expression, chondrocytes were transfected with miRNA-140 antagomir (antago miR-140), and cells that were transfected with negative control miRNA (antago miR-NC) served as a control. (a) The expression of miR-140 was decreased in antagomir-treated chondrocytes. (b–e) The mRNA levels of matrix-degrading enzyme genes, including ADAMTS4 (b), ADAMTS5 (c), MMP9 (d), and MMP13 (e), were quantified with real-time PCR using GAPDH for normalization. (f) Protein levels of matrix-degrading enzymes in melatonin-treated cells were determined using Western blot assays. Values are the of four independent experiments () in PCR experiments and three independent experiments () in Western blot assays. Statistically significant differences are indicated by # where or ## where vs. the CTRL group and where or where between the indicated groups.

(a)

(b)

(c)

(d)

(e)

(f)