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

MiR-210 targeted DR6 in chondrocytes.(a) mRNA levels of DR6 in chondrocytes with different treatment; (b) Protein levels of DR6 in chondrocytes with different treatment; (c) prediction result of miR-210 targeting DR6 3′-UTR; (d) relative luciferase activity of 293T cells with different treatment. Data are represented 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

f3: MiR-210 targeted DR6 in chondrocytes.(a) mRNA levels of DR6 in chondrocytes with different treatment; (b) Protein levels of DR6 in chondrocytes with different treatment; (c) prediction result of miR-210 targeting DR6 3′-UTR; (d) relative luciferase activity of 293T cells with different treatment. Data are represented as mean ± SD of three experimental replicates. *P < 0.05 vs. control group, #P < 0.05 vs. LPS group.

Mentions: To investigate the molecular mechanism of miR-210, the potential targets of miR-210 was predicted from RegRNA website. DR6 was finally selected as the most likely target of miR-210 during inflammation (Fig. 3c). The mRNA and protein levels of DR6 were detected by RT-PCR and western blotting, respectively. As shown in Fig. 3a,b, protein level of DR6 was increased by LPS and inhibited by miR-210 mimic. However, the change of DR6 mRNA level was not obvious, indicating that the regulation of DR6 expression was posttranscriptional gene silencing (PTGS). To confirm whether miR-210 was targeted at the 3′-UTR of DR6, a relative luciferase activity assay was performed. The relative luciferase activity significantly decreased when the cells were transfected with the wide type of DR6 3′-UTR and miR-210 mimic (Fig. 3d). The results indicated that DR6 was the direct target of miR-210.


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

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

MiR-210 targeted DR6 in chondrocytes.(a) mRNA levels of DR6 in chondrocytes with different treatment; (b) Protein levels of DR6 in chondrocytes with different treatment; (c) prediction result of miR-210 targeting DR6 3′-UTR; (d) relative luciferase activity of 293T cells with different treatment. Data are represented 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

f3: MiR-210 targeted DR6 in chondrocytes.(a) mRNA levels of DR6 in chondrocytes with different treatment; (b) Protein levels of DR6 in chondrocytes with different treatment; (c) prediction result of miR-210 targeting DR6 3′-UTR; (d) relative luciferase activity of 293T cells with different treatment. Data are represented as mean ± SD of three experimental replicates. *P < 0.05 vs. control group, #P < 0.05 vs. LPS group.
Mentions: To investigate the molecular mechanism of miR-210, the potential targets of miR-210 was predicted from RegRNA website. DR6 was finally selected as the most likely target of miR-210 during inflammation (Fig. 3c). The mRNA and protein levels of DR6 were detected by RT-PCR and western blotting, respectively. As shown in Fig. 3a,b, protein level of DR6 was increased by LPS and inhibited by miR-210 mimic. However, the change of DR6 mRNA level was not obvious, indicating that the regulation of DR6 expression was posttranscriptional gene silencing (PTGS). To confirm whether miR-210 was targeted at the 3′-UTR of DR6, a relative luciferase activity assay was performed. The relative luciferase activity significantly decreased when the cells were transfected with the wide type of DR6 3′-UTR and miR-210 mimic (Fig. 3d). The results indicated that DR6 was the direct target of miR-210.

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