Nikolaou Georgios, Zibis Aristeidis H, Fyllos Apostolos H, Katsioulis Antonios, Sotiriou Sotirios, Kotrotsios Anastasios, Sgantzos Markos, Vassiou Aikaterini, Arvanitis Dimitrios L
Department of Anatomy, Faculty of Medicine, University of Thessaly, Larissa, Greece.
Department of Hygiene and Epidemiology, Faculty of Medicine, University of Thessaly, Larissa, Greece.
Asian Spine J. 2017 Dec;11(6):863-869. doi: 10.4184/asj.2017.11.6.863. Epub 2017 Dec 7.
Human herniated discs were obtained from discectomy specimens for the immunohistochemical detection of O-GlcNAc and O-GlcNAcase (OGA)/O-GlcNAc transferase (OGT).
This study aimed to quantify the extent of O-GlcNAcylation and its associated enzymes (OGT/OGA) in human degenerated intervertebral discs.
The O-GlcNAcylation of nuclear, cytoplasmic, and mitochondrial proteins as well as the effects of such post-translational modifications are currently the focus of extensive research. O-GlcNAcylation is believed to contribute to the etiology of chronic illnesses by acting as a nutrient and stress sensor in the cellular environment. Mature intervertebral disc cells are chondrocyte-like cells, and O-GlcNAc has been shown to promote chondrocyte apoptosis . We believe that O-GlcNAcylation is a key regulator of disc degeneration.
Fifty-six specimens were fixed for 24 hours in a 10% solution of neutral-buffered formaldehyde, dehydrated, and embedded in paraffin. Tissue slices (4-µm-thick) were used for hematoxylin-eosin staining and immunohistochemistry.
We found that O-GlcNAcylation of cytoplasmic proteins was less than that of nuclear proteins in both single cells and cell clusters. Cytoplasmic O-GlcNAcylation occurs subsequent to nuclear O-GlcNAcylation and is directly proportional to disc degeneration. OGT and O-GlcNAc expression levels were identical in all specimens examined.
O-GlcNAc and OGA/OGT expression is shown to correlate for the first time with intervertebral disc cell degeneration. Increasing disc degeneration is associated with increasing O-GlcNAcylation in both nuclear and cytoplasmic proteins in human disc cells.
从椎间盘切除术标本中获取人类椎间盘，用于O-连接的N-乙酰葡糖胺（O-GlcNAc）和O-GlcNAcase（OGA）/O-GlcNAc转移酶（OGT）的免疫组织化学检测。
本研究旨在量化人类退变椎间盘中O-GlcNAc糖基化及其相关酶（OGT/OGA）的程度。
细胞核、细胞质和线粒体蛋白的O-GlcNAc糖基化以及这种翻译后修饰的作用目前是广泛研究的焦点。O-GlcNAc糖基化被认为通过在细胞环境中充当营养和应激传感器，对慢性疾病的病因学有影响。成熟的椎间盘细胞是软骨样细胞，并且已表明O-GlcNAc可促进软骨细胞凋亡。我们认为O-GlcNAc糖基化是椎间盘退变的关键调节因子。
56个标本在10%中性缓冲甲醛溶液中固定24小时，脱水并包埋于石蜡中。4微米厚的组织切片用于苏木精-伊红染色和免疫组织化学。
我们发现，在单个细胞和细胞簇中，细胞质蛋白的O-GlcNAc糖基化均少于细胞核蛋白。细胞质O-GlcNAc糖基化在细胞核O-GlcNAc糖基化之后发生，并且与椎间盘退变直接相关。在所检查的所有标本中，OGT和O-GlcNAc的表达水平相同。
首次表明O-GlcNAc和OGA/OGT的表达与椎间盘细胞退变相关。在人类椎间盘细胞中，椎间盘退变加剧与细胞核和细胞质蛋白中O-GlcNAc糖基化增加有关。
