Limits...
MiR-210 inhibits NF-κB signaling pathway by targeting DR6 in osteoarthritis.

Zhang D, Cao X, Li J, Zhao G - Sci Rep (2015)

Bottom Line: In the in vitro study, miR-210 level in chondrocytes was decreased after treatment with lipopolysaccharide (LPS).MiR-210 expression is reduced in OA rats.The results demonstrated that miR-210 decreased inflammation in articular cavity in OA rats by targeting DR6 and inhibiting NF-κB signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

ABSTRACT
Osteoarthritis (OA) is characterized by degradation of articular cartilage and joint inflammation. MicroRNAs have been proved to play an important role in the regulation of chondrogenesis. Previous study showed that microRNA-210 (miR-210) was probably associated with osteoarthritis, while the function of miR-210 in osteoarthritis still remains unknown. The aim of the present study was to investigate the protective effect of miR-210 on osteoarthritis. In the in vitro study, miR-210 level in chondrocytes was decreased after treatment with lipopolysaccharide (LPS). Transfection with miR-210 mimic inhibited LPS-induced pro-inflammatory cytokines production, cell viability reduction and cell apoptosis. Results of luciferase activity assay showed that miR-210 targeted 3'-UTR of death receptor 6 (DR6) to inhibit its expression. MiR-210 mimic and DR6 siRNA transfection inhibited the activation of NF-κB pathway and cell apoptosis of chondrocytes. For the in vivo study, OA model was established on rats by anterior cruciate ligament transection (ACLT). MiR-210 expression is reduced in OA rats. MiR-210 over-expressing lentivirus was injected into the OA rats. Cytokines production, and NF-κB and DR6 expression in OA rats was inhibited by miR-210 overexpression. The results demonstrated that miR-210 decreased inflammation in articular cavity in OA rats by targeting DR6 and inhibiting NF-κB signaling pathway.

No MeSH data available.


Related in: MedlinePlus

Effect of miR-210 on LPS-induced chondrocytes.(a) Production of IL-1β, IL-6 and TNF-α in the cell culture supernatants was determined by ELISA; (b) Cell viability was detected by MTT assay; (c–d) Cell apoptosis was measured by FCM. Data are shown as mean ± SD of three experimental replicates. *P<0.05 vs. control group; #P < 0.05 vs. LPS group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4525484&req=5

f2: Effect of miR-210 on LPS-induced chondrocytes.(a) Production of IL-1β, IL-6 and TNF-α in the cell culture supernatants was determined by ELISA; (b) Cell viability was detected by MTT assay; (c–d) Cell apoptosis was measured by FCM. Data are shown as mean ± SD of three experimental replicates. *P<0.05 vs. control group; #P < 0.05 vs. LPS group.

Mentions: LPS usually induces inflammation and causes cell apoptosis. After incubation with LPS, levels of IL-1β, IL-6 and TNF-α in cell supernatant were detected by ELISA. Results showed that LPS induced secretion of IL-1β, IL-6 and TNF-α (Fig. 2a). The levels of IL-1β and TNF-α were decreased in miR-210 mimic transfected cells (Fig. 2a). The cell viability of chondrocytes was detected by MTT assay. As shown in Fig. 2b, the relative cell viability was decreased by 55% after the treatment of LPS, whereas the cell viability was increased by 32% after miR-210 transfection. Flow cytometry was utilized to quantify LPS-induced chondrocytes apoptosis. As shown in Fig. 2c,d, the percentage of cell apoptosis was increased to 39.1% in LPS-induced cells (P < 0.01), and reduced to 25.6% after miR-210 mimic transfection (P < 0.05). The results indicated that miR-210 protect the chondrocytes from LPS induced injury.


MiR-210 inhibits NF-κB signaling pathway by targeting DR6 in osteoarthritis.

Zhang D, Cao X, Li J, Zhao G - Sci Rep (2015)

Effect of miR-210 on LPS-induced chondrocytes.(a) Production of IL-1β, IL-6 and TNF-α in the cell culture supernatants was determined by ELISA; (b) Cell viability was detected by MTT assay; (c–d) Cell apoptosis was measured by FCM. Data are shown as mean ± SD of three experimental replicates. *P<0.05 vs. control group; #P < 0.05 vs. LPS group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525484&req=5

f2: Effect of miR-210 on LPS-induced chondrocytes.(a) Production of IL-1β, IL-6 and TNF-α in the cell culture supernatants was determined by ELISA; (b) Cell viability was detected by MTT assay; (c–d) Cell apoptosis was measured by FCM. Data are shown as mean ± SD of three experimental replicates. *P<0.05 vs. control group; #P < 0.05 vs. LPS group.
Mentions: LPS usually induces inflammation and causes cell apoptosis. After incubation with LPS, levels of IL-1β, IL-6 and TNF-α in cell supernatant were detected by ELISA. Results showed that LPS induced secretion of IL-1β, IL-6 and TNF-α (Fig. 2a). The levels of IL-1β and TNF-α were decreased in miR-210 mimic transfected cells (Fig. 2a). The cell viability of chondrocytes was detected by MTT assay. As shown in Fig. 2b, the relative cell viability was decreased by 55% after the treatment of LPS, whereas the cell viability was increased by 32% after miR-210 transfection. Flow cytometry was utilized to quantify LPS-induced chondrocytes apoptosis. As shown in Fig. 2c,d, the percentage of cell apoptosis was increased to 39.1% in LPS-induced cells (P < 0.01), and reduced to 25.6% after miR-210 mimic transfection (P < 0.05). The results indicated that miR-210 protect the chondrocytes from LPS induced injury.

Bottom Line: In the in vitro study, miR-210 level in chondrocytes was decreased after treatment with lipopolysaccharide (LPS).MiR-210 expression is reduced in OA rats.The results demonstrated that miR-210 decreased inflammation in articular cavity in OA rats by targeting DR6 and inhibiting NF-κB signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

ABSTRACT
Osteoarthritis (OA) is characterized by degradation of articular cartilage and joint inflammation. MicroRNAs have been proved to play an important role in the regulation of chondrogenesis. Previous study showed that microRNA-210 (miR-210) was probably associated with osteoarthritis, while the function of miR-210 in osteoarthritis still remains unknown. The aim of the present study was to investigate the protective effect of miR-210 on osteoarthritis. In the in vitro study, miR-210 level in chondrocytes was decreased after treatment with lipopolysaccharide (LPS). Transfection with miR-210 mimic inhibited LPS-induced pro-inflammatory cytokines production, cell viability reduction and cell apoptosis. Results of luciferase activity assay showed that miR-210 targeted 3'-UTR of death receptor 6 (DR6) to inhibit its expression. MiR-210 mimic and DR6 siRNA transfection inhibited the activation of NF-κB pathway and cell apoptosis of chondrocytes. For the in vivo study, OA model was established on rats by anterior cruciate ligament transection (ACLT). MiR-210 expression is reduced in OA rats. MiR-210 over-expressing lentivirus was injected into the OA rats. Cytokines production, and NF-κB and DR6 expression in OA rats was inhibited by miR-210 overexpression. The results demonstrated that miR-210 decreased inflammation in articular cavity in OA rats by targeting DR6 and inhibiting NF-κB signaling pathway.

No MeSH data available.


Related in: MedlinePlus