特发性脊柱侧凸椎旁肌组织钙调蛋白和神经元型一氧化氮合酶表达的研究

目的通过比较特发性脊柱侧凸患者胸弯顶点处凸侧、凹侧椎旁肌的钙调蛋白(CaM)和神经元型一氧化氮合酶(nNOS)表达的差异,探讨椎旁肌在脊柱侧凸中可能的作用机制。方法本研究选择了2001年8-12月期间北京协和医院收治的胸椎侧凸畸形患者10例,手术中取脊柱侧凸患者顶椎凸侧、凹侧椎旁肌肉组织各1块,采用免疫组织化学和Western杂交检测脊柱侧凸椎旁肌组织中CaM和nNOS基因的表达,采用HE染色和SA染色检测椎旁肌组织肌纤维形态。结果HE染色和SA染色显示脊柱侧凸患者凸侧肌纤维比其凹侧及正常对照的肌纤维明显变细。免疫组织化学和Western杂交显示凸侧肌肉组织CaM和nNOS的表达量明显降低。结论脊柱侧凸患者双侧椎旁肌CaM和nNOS表达不均衡,可能在特发性脊柱侧凸发生发展中发挥作用。

Expression of calmodulin and nNOS in the paraspinal muscles in idiopathic scoliosis

Objective The aim of this study was to identify whether one side of the apex of the scoliotic curve showed greater muscular abnormalities than the other. This investigation sought to compare directly the expression and localization of Calmodulin (CaM) and nNOS of the thoracic spinal musculature on convex side with those on the concave side in the scoliosis patients attending for spinal surgery. Methods The patient group comprised 8 females and 2 males, scheduled for spinal surgery, in whom the apex of the scoliotic curve arose between T6 and T11. Muscle biopsies were taken bilaterally during surgery from the superficial multifidus muscle at the apex of the curve between the 6th and 11 th thoracic vertebral levels. Part of the tissue was fixed in Formalin and stained with hematoxylin and eosin; the remaining tissue was snap frozen and processed for immunohistochemistry and western blot. The following antibodys were used for the immunohistochemistry at dilutions optimized in preliminary experiments. Goat polyclonal anti-human-CaM and mouse monoclonal anti-human-nNOS were used as primary antibodys. CaM and nNOS were detected by immunohistochemical method and Western-blot. Western blotting was done with antibodys to CaM. Results Compared with control muscle and the muscle on the concave side of the scoliotic curve, A significant decrease in CaM and nNOS was found on the convex side of paraspinal muscle from idiopathic scoliosis patients, Western blotting with an CaM antibody and nNOS antibody shows that there are more CaM and nNOS protein in the concave side of paraspinal muscle in these patients with adolescent idiopathic scoliosis than in the convex side. Conclusion This asymmetry of CaM and nNOS in the paraspinal muscle may play a role in the pathogenensis of IS.