Shen Lin, Wu Yinghua, Han Liang, Zhang Haiying
Graduate School, Tianjin Medical University, Tianjin 300070, P.R. China.
Department of Orthopedic Trauma, Tianjin Hospital, Tianjin 300211, P.R. China.
Exp Ther Med. 2018 Apr;15(4):3603-3608. doi: 10.3892/etm.2018.5867. Epub 2018 Feb 14.
Low back pain (LBP) is one of the most common musculoskeletal diseases in the world. The incidence is ~70% in adults and many of them suffer from disability. Recently, intervertebral disc degeneration (IDD) has been deemed as a main cause of LBP. The present study aimed to investigate the potentials of growth and differentiation factor-5 (GDF-5) in IDD. The protein levels of prostaglandin-E2 (PGE2), tumor necrosis factor (TNF)-α and interleukin (IL)-1β in culture medium were evaluated by ELISA. mRNA and protein expression levels in nucleus pulposus (NP) cells were evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. Griess reaction was applied to test the nitric oxide (NO) concentration in the culture supernatant. The expression levels of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in NP cells were measured by RT-qPCR. Collagen-II, aggrecan, IκBα and phosphorylated (p)-p65 expression levels were detected by western blotting. Compared with the control group, protein expression levels of TNF-α, IL-1β and PGE2, and NO concentration in culture medium were upregulated by LPS, which were significantly repressed by GDF-5 overexpression (P<0.05). Additionally, GDF-5 overexpression reduced lipopolysaccharide-induced upregulation of TNF-α, IL-1β, iNOS, COX-2, collagen-II, aggrecan, IκBα and p-p65 expression levels in NP cells.
下腰痛(LBP)是世界上最常见的肌肉骨骼疾病之一。成年人中的发病率约为70%,其中许多人患有残疾。最近,椎间盘退变(IDD)被认为是LBP的主要原因。本研究旨在探讨生长分化因子5(GDF-5)在IDD中的作用潜力。采用酶联免疫吸附测定(ELISA)法评估培养基中前列腺素E2(PGE2)、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β的蛋白水平。分别采用逆转录定量聚合酶链反应(RT-qPCR)和蛋白质印迹法评估髓核(NP)细胞中的mRNA和蛋白表达水平。采用格里斯反应检测培养上清液中的一氧化氮(NO)浓度。采用RT-qPCR法检测NP细胞中诱导型NO合酶(iNOS)和环氧化酶-2(COX-2)的表达水平。采用蛋白质印迹法检测Ⅱ型胶原蛋白、聚集蛋白聚糖、IκBα和磷酸化(p)-p65的表达水平。与对照组相比,脂多糖(LPS)上调了培养基中TNF-α、IL-1β和PGE2的蛋白表达水平以及NO浓度,而GDF-5过表达可显著抑制这些指标(P<0.05)。此外,GDF-5过表达降低了脂多糖诱导的NP细胞中TNF-α、IL-1β、iNOS、COX-2、Ⅱ型胶原蛋白、聚集蛋白聚糖、IκBα和p-p65表达水平的上调。