Ma Mei, Liang Xiao, Wang Xi, Zhang Lu, Cheng Shiqiang, Guo Xiong, Zhang Feng, Wen Yan
School of Public Health, Health Science Center, Xi'an Jiaotong University, Xi'an, China.
National Local Joint Engineering Research Center of Biodiagnostics and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Bone Joint Res. 2020 Sep 20;9(9):578-586. doi: 10.1302/2046-3758.99.BJR-2019-0247.R1. eCollection 2020 Sep.
Kashin-Beck disease (KBD) is a kind of chronic osteochondropathy, thought to be caused by environmental risk factors such as T-2 toxin. However, the exact aetiology of KBD remains unclear. In this study, we explored the functional relevance and biological mechanism of cartilage oligosaccharide matrix protein (COMP) in the articular cartilage damage of KBD.
The articular cartilage specimens were collected from five KBD patients and five control subjects for cell culture. The messenger RNA (mRNA) and protein expression levels were detected by quantitative reverse transcription PCR (qRT-PCR) and western blot. The survival rate of C28/I2 chondrocyte cell line was detected by MTT assay after T-2 toxin intervention. The cell viability and mRNA expression levels of apoptosis related genes between -overexpression groups and control groups were examined after cell transfection.
The mRNA and protein expression levels of were significantly lower in KBD chondrocytes than control chondrocytes. After the T-2 toxin intervention, the mRNA expression of C28/I2 chondrocyte reduced and the protein level of COMP in three intervention groups was significantly lower than in the control group. MTT assay showed that the survival rate of overexpression KBD chondrocytes were notably higher than in the blank control group. The mRNA expression levels of , , and were also significantly different among COMP overexpression, negative control, and blank control groups.
Our study results confirmed the functional relevance of with KBD. may play an important role in the excessive chondrocytes apoptosis of KBD patients.Cite this article: 2020;9(9):578-586.
大骨节病（KBD）是一种慢性骨软骨病，被认为是由T-2毒素等环境危险因素引起的。然而，KBD的确切病因仍不清楚。在本研究中，我们探讨了软骨寡糖基质蛋白（COMP）在KBD关节软骨损伤中的功能相关性及生物学机制。
收集5例KBD患者和5例对照者的关节软骨标本进行细胞培养。通过定量逆转录PCR（qRT-PCR）和蛋白质免疫印迹法检测信使核糖核酸（mRNA）和蛋白质表达水平。T-2毒素干预后，采用MTT法检测C28/I2软骨细胞系的存活率。细胞转染后检测过表达组与对照组之间细胞活力及凋亡相关基因的mRNA表达水平。
KBD软骨细胞中COMP的mRNA和蛋白质表达水平显著低于对照软骨细胞。T-2毒素干预后，C28/I2软骨细胞的COMP mRNA表达降低，三个干预组的COMP蛋白水平均显著低于对照组。MTT法检测显示，COMP过表达KBD软骨细胞的存活率显著高于空白对照组。COMP过表达组、阴性对照组和空白对照组之间，Bcl-2、Bax、Caspase-3和Caspase-9的mRNA表达水平也存在显著差异。
我们的研究结果证实了COMP与KBD的功能相关性。COMP可能在KBD患者软骨细胞过度凋亡中起重要作用。引用本文： 2020；9(9):578-586。
