Abstract：Objective: To study the regulatory effect of LncRNA uc.48+ on inflammation of knee osteoarthritis (OA) and explore its mechanism. Methods: Sodium iodoacetate (MIA) was injected into the joint cavity to induce OA in SD rats. The mechanical withdrawal threshold (MWT) and paw withdrawal thermal latency (PWTL) on the injection side of the knee joint were recorded within 21 days after injection. OS staining was used to observe the pathological changes of 21d joint tissues, and real-time quantitative PCR (qRT-PCR) was used to detect the levels of cartilage LncRNA uc.48+ and P2X7R. Plasmid pcDNA3.1 was used to construct LncRNA uc.48+ overexpression vector (pc-LncRNA uc.48+) and empty vector (pc-null). The rats were divided into sham group (n=10) and MIA group (n=50). Among them, 40 rats were randomly selected from the MIA group and randomly divided into pc-null group, pc-LncRNAuc.48+ group, pc-LncRNA uc .48++ Nucleotide P2X receptor (P2X7R) channel antagonist (AZD9056) group, pc-LncRNA uc.48++ nuclear factor κB (NF-κB) inhibitor (Helenalin) group (n=10). Western blot was used to detect the levels of P2X7R, P65, phosphorylated P65 (p-P65), matrix metalloproteinase (MMP)-13 and substance P (SP) in the above groups. ELISA was used to detect cartilage tissue prostaglandin (PG) E2, tumor necrosis factor (TNF)-α, and interleukin (IL)-1β. In vitro chondrocytes were transfected with pc-LncRNA uc.48+ or pc-null for 24h to detect the expression of P2X7R, P65, and p-P65. Results: Compared with the pre-modeling, MWT and PWTL of rats after 1d-21d of MIA injection decreased, and the joint PGE2, TNF-α, IL-1β all increased (all P<0.05). LncRNA uc.48+ continued to be up-regulated from 1d to 21d in the MIA group, and P2X7R mRNA was also up-regulated (all P<0.05). In MIA rats, the levels of PGE2, TNF-α, IL-1β, P2X7R, P65, and p-P65 in the pc-LncRNAuc. 48+ group were higher than those in the pc-null group (all P<0.05), while the levels of PGE2, TNF-α, IL-1β, P65, p-P65, MMP-13 and SP in the uc.48++AZD9056 group were lower than those in the pc-LncRNAuc.48+ group (all P<0.05). In addition, the levels of PGE2, TNF-α, IL-1β, MMP-13 and SP in the pc-LncRNA uc.48++ Helenalin group were lower than those in the pc-LncRNAuc.48+ group (all P<0.05). However, the P2X7R level of the pc-LncRNA uc.48++ Helenalin group and the pc-LncRNAuc.48+ group were not different (P>0.05). Conclusion: LncRNA uc.48+ promotes the inflammation of osteoarthritis cartilage tissue by activating the P2X7R/NF-κB signaling pathway.
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2022, Vol. 28 
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