Xie Wei, Jiang Luoyong, Huang Xiaoyang, Shang Hongxi, Gao Minghong, You Wei, Tan Jifeng, Yan Hong, Sun Wei
Department of Orthopedics, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, Shenzhen, Guangdong 518000, P.R. China.
Exp Ther Med. 2021 Oct;22(4):1153. doi: 10.3892/etm.2021.10587. Epub 2021 Aug 10.
Long noncoding RNA (lncRNA) maternally expressed 8, small nucleolar RNA host gene (MEG8) has been widely reported for its pro-proliferative, anti-apoptotic and anti-inflammatory effects in diverse diseases. The aim of the present study was to investigate the effects and underlying mechanism of MEG8 on IL-1β-stimulated human osteoarthritis (OA) chondrocytes. C28/I2 chondrocytes were cultured under the stimulation of IL-1β to establish a cellular model of OA. Functional assays involving Cell Counting Kit-8 and flow cytometry were performed to determine proliferation and apoptosis in the cells. The protein expression levels of caspase-3 and inflammatory cytokines were detected using cell-based ELISA. The expression levels of PI3K/AKT pathway-related proteins were evaluated by western blotting. It was identified that MEG8 expression was increased in the cartilage of patients with OA and in IL-1β-treated C28/I2 cells. In C28/I2 cells, silencing of MEG8 expression noticeably triggered IL-1β-induced proliferation suppression, cell death and an inflammatory response. However, transfection with MEG8 displayed adverse effects. Furthermore, MEG8 overexpression prevented IL-1β-induced activation of the PI3K/AKT signaling pathway in C28/I2 cells. These data demonstrated that MEG8 exerted protective effects against IL-1β-induced apoptosis and inflammation of OA chondrocytes by regulating the PI3K/AKT signaling pathway. Thus, the present study demonstrates that MEG8 might be a promising target for the treatment of OA.
长链非编码RNA(lncRNA)母系表达8,即小核仁RNA宿主基因(MEG8),其在多种疾病中的促增殖、抗凋亡和抗炎作用已被广泛报道。本研究旨在探讨MEG8对白细胞介素-1β(IL-1β)刺激的人骨关节炎(OA)软骨细胞的影响及潜在机制。在IL-1β刺激下培养C28/I2软骨细胞以建立OA细胞模型。进行了涉及细胞计数试剂盒-8和流式细胞术的功能测定,以确定细胞中的增殖和凋亡情况。使用基于细胞的酶联免疫吸附测定法检测半胱天冬酶-3和炎性细胞因子的蛋白表达水平。通过蛋白质印迹法评估PI3K/AKT通路相关蛋白的表达水平。研究发现,OA患者软骨组织及经IL-1β处理的C28/I2细胞中MEG8表达增加。在C28/I2细胞中,沉默MEG8表达显著引发IL-1β诱导的增殖抑制、细胞死亡和炎症反应。然而,转染MEG8则产生相反作用。此外,MEG8过表达可阻止IL-1β诱导的C28/I2细胞中PI3K/AKT信号通路激活。这些数据表明,MEG8通过调节PI3K/AKT信号通路对IL-1β诱导的OA软骨细胞凋亡和炎症发挥保护作用。因此,本研究表明MEG8可能是治疗OA的一个有前景的靶点。