Long non-coding RNA NORAD regulates IL-1β-induced inflammatory injury in osteoarthritis chondrocytes by regulating the NF-κB signaling pathway
Objective To explore the role of long non-coding RNA NORAD(lncRNA NORAD)in IL-1β-induced inflammatory injury of osteoarthritis chondrocytes and the mechanism.Methods An in vitro osteoarthritis model was established by IL-1β induction in chondrocytes.The chondrocytes were divided into four groups,including control group,IL-1β group(stimulation of chondrocytes with 10ng/mL of IL-1β for 24h),IL-1β + si-NC group(transfection with siRNA NC for 48h followed by IL-1β stimulation),and IL-1β + si-NORAD group(transfection with NORAD siRNA for 48 h followed by IL-1β stimulation).The expression level of NORAD was detected by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR).The survival rate of chondrocytes was measured using the Calcein AM Cell Viability Assay Kit.The apoptotic level of chondrocytes was assessed by the One Step TUNEL Apoptosis Assay Kit.The protein levels of Bax,B-cell lymphoma 2(Blc-2),metalloproteinase 13(MMP-13),collagen typeⅡ alpha 1 chain(COL2A1),and nuclear factor kappa B(NF-κB)p65 were examined by Western blot.The concentrations of interleukin 6(IL-6)and tumor necrosis factor-alpha(TNF-α)were determined by enzyme-linked immunosorbent assay(ELISA).Results Compared with that of the control group,NORAD was significantly upregulated in IL-1β group(P<0.01).Compared with that of the IL-1β + si-NC group,NORAD was significantly downregulated in IL-1β + si-NORAD group(P<0.01).Compared with those of the control group,significantly lower survival,and significantly higher apoptotic rate,degradation of extracellular matrix,concentrations of IL-6 and TNF-α,and phosphorylated NF-κB p65 level were detected in IL-1β group and IL-1β + si-NC group(all P<0.01).Compared with those of the IL-1β group and IL-1β + si-NC group,significantly higher survival,and significantly lower apoptotic rate,degradation of extracellular matrix,concentrations of IL-6 and TNF-α,and phosphorylated NF-κB p65 level were detected in the chondrocytes of IL-1β + si-NORAD group(all P<0.01).Conclusion Knockdown of lncRNA NORAD ameliorates IL-1β-induced inflammatory injury of chondrocytes by inhibiting the NF-κB signaling pathway.
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维普
