Novosibirsk Research Institute of Traumatology and Orthopaedics n.a. Ya.L. Tsivyan, Novosibirsk, Russia.
Meshalkin National Medical Research Center, Ministry of Health of the Russian Federation, Novosibirsk, Russia.
Int J Med Sci. 2019 Jan 1;16(2):221-230. doi: 10.7150/ijms.29312. eCollection 2019.
In a previous report, we demonstrated the presence of cells with a neural/glial phenotype on the concave side of the vertebral body growth plate in Idiopathic Scoliosis (IS) and proposed this phenotype alteration as the main etiological factor of IS. In the present study, we utilized the same specimens of vertebral body growth plates removed during surgery for Grade III-IV IS to analyse gene expression. We suggested that phenotype changes observed on the concave side of the vertebral body growth plate can be associated with altered expression of particular genes, which in turn compromise mechanical properties of the concave side. We used a Real-Time SYBR Green PCR assay to investigate gene expression in vertebral body growth plates removed during surgery for Grade III-IV IS; cartilage tissues from human fetal spine were used as a surrogate control. Special attention was given to genes responsible for growth regulation, chondrocyte differentiation, matrix synthesis, sulfation and transmembrane transport of sulfates. We performed morphological, histochemical, biochemical, and ultrastructural analysis of vertebral body growth plates Expression of genes that control chondroitin sulfate sulfation and corresponding protein synthesis was significantly lower in scoliotic specimens compared to controls. Biochemical analysis showed 1) a decrease in diffused proteoglycans in the total pool of proteoglycans; 2) a reduced level of their sulfation; 3) a reduction in the amount of chondroitin sulfate coinciding with raising the amount of keratan sulfate; and 4) reduced levels of sulfation on the concave side of the scoliotic deformity. The results suggested that altered expression of genes that control chondroitin sulfate sulfation and corresponding changes in protein synthesis on the concave side of vertebral body growth plates could be causal agents of the scoliotic deformity.
在之前的报告中，我们证明了特发性脊柱侧凸（IS）的椎体生长板凹侧存在具有神经/胶质表型的细胞，并提出这种表型改变是 IS 的主要病因因素。在本研究中，我们利用相同的 IS Ⅲ-Ⅳ级手术切除的椎体生长板标本进行基因表达分析。我们提出，在椎体生长板凹侧观察到的表型变化可能与特定基因表达的改变有关，这反过来又损害了凹侧的机械性能。我们使用实时 SYBR Green PCR 检测法研究了 IS Ⅲ-Ⅳ级手术切除的椎体生长板中的基因表达；用人胎儿脊柱软骨组织作为替代对照。特别关注负责生长调节、软骨细胞分化、基质合成、硫酸化和硫酸盐跨膜转运的基因。我们对椎体生长板进行了形态学、组织化学、生物化学和超微结构分析。与对照组相比，脊柱侧凸标本中控制硫酸软骨素硫酸化和相应蛋白合成的基因表达显著降低。生化分析显示：1）在总蛋白聚糖池中扩散型蛋白聚糖减少；2）硫酸化水平降低；3）硫酸软骨素减少，而角蛋白硫酸增加；4）脊柱侧凸畸形凹侧硫酸化水平降低。结果表明，控制硫酸软骨素硫酸化的基因表达改变以及椎体生长板凹侧相应的蛋白合成变化可能是脊柱侧凸畸形的原因。
