Ⅰ型神经纤维瘤病软骨细胞生物学特性探究.docVIP

Ⅰ型神经纤维瘤病软骨细胞生物学特性探究 　 作者：陈晖，邱勇，王斌，俞扬，朱泽章，朱锋，钱邦平，马薇薇 【摘要】 　　目的 检测神经纤维瘤蛋白在Ⅰ型神经纤维瘤病(type 1 neurofibromatosis，NF1)脊柱侧凸患者软骨细胞中的表达，观察NF1脊柱侧凸患者软骨细胞的生物学特性。方法 先天性脊柱侧凸患者7 例，NF1脊柱侧凸患者6 例，两组年龄和Cobb角接近。脊柱后路手术中取髂骨生长板行软骨细胞分离和培养。取第2代软骨细胞分别检测其增殖活性和Ⅱ型胶原、可聚蛋白多糖等软骨细胞特异性分化指标，并用免疫沉淀和Western blot检测神经纤维瘤蛋白在两组软骨细胞中的表达。结果 NF1脊柱侧凸患者软骨细胞中神经纤维瘤蛋白表达水平明显低于先天性脊柱侧凸组(灰度积分比值分别为1.31±0.53和2.17±1.02，P＝0.03)，其Ⅱ型胶原水平明显低于先天性脊柱侧凸患者组(26.6±11.3 ng/mgpro 和31.7±16.2 ng/mgpro，P＝0.03)，而可聚蛋白多糖水平两组无明显差异(63.0±129.6 ng/mgpro和76.7±519.4 ng/mgpro，P＝0.21)。NF1脊柱侧凸患者软骨细胞显示相对活跃的增殖活性(增殖倍数分别为2.9±0.04和2.49±0.11，P＝0.02)。结论 NF1脊柱侧凸患者软骨细胞神经纤维瘤蛋白表达降低的同时，其细胞增值相对活跃，细胞分化功能也存在缺陷。 【关键词】 神经纤维瘤病 脊柱侧凸 神经纤维瘤蛋白 软骨细胞 生物学特性 　　Biological Features of Chondrocytes Isolated from Patients with Type Ⅰ Neurofibromatosis 　　Abstract：Objective To detect the neurofibromin expression and observe the biological features of the chondrocytes of patients with type Ⅰneurofibromatosis(NF1) and scoliosis.Methods 7 cases of congenital scoliosis and 6 cases of NF1 scoliosis were chosen with the similar age and Cobb angle.The growth plate was harvested from the ilia，diced and sequentially digested with enzymes.Proliferation of the chondrocytes，and also the specific differentiation indice including type Ⅱ collagen and aggrecan were assayed.Neurofibromin expression in the two kinds of chondrocytes was detected with immunoprecipitation followed by Western blot.Results Compared to the chondrocytes of patients with congenital scoliosis，lower level of neurofibromin was expressed in chondrocytes of patients with NF1 scoliosis(the OD value was 1.31±0.53 and 2.17±1.02 respectively，P＝0.03).The level of type Ⅱ collagen was lower in chondrocytes of patients with NF1 scoliosis(26.6±11.3 ng/mgpro vs 31.7±16.2 ng/mgpro，P＝0.03)，but the level of aggrecan does not differ between the two groups(63.0±129.6 ng/mgpro和76.7±519.4 ng/mgpro，P＝0.21).The proliferation rate of the NF1 chondrocytes was higher(2.9±0.04 and 2.49±0.11，P＝0.02).Conclusion With the decrease of neurofibromin expre