Zhou Zhenggang, Wang Yini, Liu Haifei, Wang Lu, Liu Zonghan, Yuan Huimei, Liu Lantao, Guo Mingbo, Wang Dechun
Medical College, Qingdao University, Qingdao, Shandong, China.
Department of Emergency Medicine, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Connect Tissue Res. 2021 Jul;62(4):359-368. doi: 10.1080/03008207.2020.1743697. Epub 2020 Mar 30.
: Intervertebral disc (IVD) degeneration (IDD) is one of the main causes for spinal degenerative diseases, such as disk herniation, spinal canal stenosis, and spinal deformities. Growing evidence has highlighted the contribution of oxidative stress in pathogenesis of IDD, and antioxidant treatment is thus considered to be a promising therapeutic strategy for IDD. The aim of this study was to investigate whether N-tert-butyl-α-phenylnitrone (PBN), a free radical scavenger, could attenuate the pathological changes of IDD by alleviating oxidative stress.: Nucleus pulposus (NP) cells were isolated from rabbit lumbar disks. MTT assay, real-time PCR and western blotting were employed to evaluate the effects of PBN on oxidative damages induced by 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) in NP cells.: AAPH induced oxidative stress and the subsequent degenerative changes in NP cells via the ERK/MAPK pathway. On the contrary, the oxidative stress induced by AAPH was significantly ameliorated by PBN. Moreover, PBN also attenuated AAPH-induced expression of matrix degradation proteases and apoptosis. PBN suppresses AAPH-induced activation of ERK/MAPK pathway, which may be the underlying mechanism for the protective effects of PBN.: Our study for the first time identified a novel role and mechanism for PBN in protecting the IVD against oxidative stress, matrix catabolism and apoptosis, which may have implications for its further application in combating IVD degenerative diseases. AAPH: 2,2'-azobis(2-methylpropanimidamidine) dihydrochloride; ADAMTS: a disintegrin and metalloproteinase with thrombospondin motifs; AF: annulus fibrosus; CEP: cartilage endplate; DCF: 2'7'-dichlorofluorescein; IDD: intervertebral disc degeneration; IVD: intervertebral disc; LPS: lipopolysaccharide; MMP: matrix metalloproteinase; MTT: methyl-thiazolyl-tetrazolium; NP: nucleus pulposus; PBN: N-tert-butyl-alfa-phenylnitrone; PGs: proteoglycans; ROS: reactive oxygen species; SDS: sodium dodecyl sulfate.
椎间盘退变（IDD）是诸如椎间盘突出、椎管狭窄和脊柱畸形等脊柱退行性疾病的主要病因之一。越来越多的证据表明氧化应激在IDD发病机制中的作用，因此抗氧化治疗被认为是一种有前景的IDD治疗策略。本研究的目的是探究自由基清除剂N-叔丁基-α-苯基硝酮（PBN）是否能通过减轻氧化应激来减轻IDD的病理变化。
从兔腰椎间盘分离出髓核（NP）细胞。采用MTT法、实时荧光定量PCR和蛋白质印迹法评估PBN对2,2'-偶氮二异丁脒二盐酸盐（AAPH）诱导的NP细胞氧化损伤的影响。
AAPH通过ERK/MAPK途径诱导NP细胞氧化应激及随后的退变变化。相反，PBN显著改善了AAPH诱导的氧化应激。此外，PBN还减轻了AAPH诱导的基质降解蛋白酶表达和细胞凋亡。PBN抑制AAPH诱导的ERK/MAPK途径激活，这可能是PBN发挥保护作用的潜在机制。
我们的研究首次确定了PBN在保护椎间盘免受氧化应激、基质分解代谢和细胞凋亡方面的新作用和机制，这可能对其在对抗椎间盘退行性疾病中的进一步应用具有重要意义。AAPH：2,2'-偶氮二异丁脒二盐酸盐；ADAMTS：含血小板反应蛋白基序的解聚素和金属蛋白酶；AF：纤维环；CEP：软骨终板；DCF：2',7'-二氯荧光素；IDD：椎间盘退变；IVD：椎间盘；LPS：脂多糖；MMP：基质金属蛋白酶；MTT：甲基噻唑基四氮唑；NP：髓核；PBN：N-叔丁基-α-苯基硝酮；PGs：蛋白聚糖；ROS：活性氧；SDS：十二烷基硫酸钠
