lncRNA ZFAS1 promotes intervertebral disc degeneration by upregulating AAK1

Abstract

We investigated the function of lncRNA zinc finger antisense 1 (ZFAS1) in intervertebral disc degeneration (IDD) progression in vitro and in vivo. Nucleus pulposus (NP) tissues were obtained from 20 patients with IDD. IL-1β was used to stimulate primary NP cells to establish the IDD models in vitro. Gene expression was determined by RT-qPCR. 5-Ethynyl-2′-deoxyuridine and flow cytometry were performed to determine cell proliferation and apoptosis, and western blotting was conducted to measure the apoptosis- and extracellular matrix (ECM)-related protein expression. Luciferase reporter assay was used to examine the interactions between the genes. We also investigated the effect of ZFAS1 in a mouse model of IDD induced by needle punctures. Our results showed that ZFAS1 expression was elevated in degenerative NP tissues and IL-1β-treated NP cells. ZFAS1 knockdown inhibited NP cell apoptosis and ECM degradation induced by IL-1β. Mechanically, ZFAS1 sponged miR-4711-5p and adaptor-associated kinase 1 (AAK1) was targeted by miR-4711-5p. Furthermore, AAK1 overexpression partially eliminated the impact of ZFAS1 depletion on NP cell proliferation, apoptosis, and ECM degradation. More importantly, the results of the in vivo studies confirmed the effect of silencing ZFAS1 on alleviating the symptoms of IDD mice. Overall, silencing ZFAS1 inhibits IDD progression by reducing NP cell apoptosis and ECM degradation through the miR-4711-5p/AAK1 axis.

Extracellular vesicles produced by avian pathogenic Escherichia coli (APEC) activate macrophage proinflammatory response and neutrophil extracellular trap (NET) formation through TLR4 signaling

“…The apoptosis of macrophages was detected with flow cytometry using an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme Biotech), according to the manufacturer’s instructions but with slight modifications [ 81 ]. Briefly, HD11 and PMA-treated THP-1 macrophages and were incubated with APEC EVs for 24 h. The cells were digested with EDTA-free trypsin.…”

Research progress on long non‑coding RNAs in non‑infectious spinal diseases (Review)

“…This previous study also revealed that ZNFX1 antisense RNA 1 (ZFAS1) was upregulated in degenerative NP tissues and NP cells treated with IL-1β, and that ZFAS1 competitively bound to miR-4711-5p through a ceRNA mechanism, thereby affecting the expression of adaptor-associated kinase 1, which is the target of miR-4711-5p. Silencing ZFAS1 may inhibit IDD progression by reducing NP apoptosis and ECM degradation ( 72 ). Similar to most lncRNAs, long intergenic nonprotein coding RNA 284 was revealed to be upregulated in IDD tissues and IL-1β-induced NP cells, and it activated the Wnt/β-catenin pathway by sponging miR-205-3p, thereby inhibiting NP cell proliferation and ECM synthesis ( 73 ).…”