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作 者:陈晖[1] 邱勇[1] 陈蕾蕾[1] 李雷[2] 周祀桥[3]
机构地区:[1]南京大学医学院附属鼓楼医院脊柱外科,南京市中山路321号210008 [2]南京大学医学院附属鼓楼医院科研部,南京市中山路321号210008 [3]南京大学医学院附属鼓楼医院病理科,南京市中山路321号210008
出 处:《中国脊柱脊髓杂志》2006年第3期220-224,i0001,共6页Chinese Journal of Spine and Spinal Cord
基 金:南京市科技攻关重点项目;项目编号:ZKX0313
摘 要:目的:检测神经纤维瘤蛋白在先天性脊柱侧凸患者成骨细胞和软骨细胞中的表达。方法:6例先天性脊柱侧凸患者,在后路手术时取髂骨及髂骨生长板,分离、培养成骨细胞和软骨细胞,分别行碱性磷酸酶染色和甲苯胺蓝染色。逆转录-多聚酶链反应(RT-PCR)检测神经纤维瘤蛋白mRNA,间接免疫荧光和Westernblot检测神经纤维瘤蛋白在成骨细胞和软骨细胞中的表达。结果:先天性脊柱侧凸患者成骨细胞和软骨细胞中存在Ⅱ型神经纤维瘤蛋白表达,该蛋白主要分布在细胞浆,所表达蛋白为三磷酸鸟苷酶活化蛋白(GAP)活性较弱的Ⅱ型异构体。结论:先天性脊柱侧凸患者成骨细胞和软骨细胞中存在神经纤维瘤蛋白表达,但该蛋白是否通过对成骨细胞和软骨细胞的影响导致骨骼系统异常还有待于进一步研究。Objective:To detect the expression of neurofibromin in osteoblasts and chondrocytes from patients with congenital scoliosis.Method:Tbe cancellous bone and growth plate were harvested from ilia of 6 patients with congenital scoliosis in posterior approach surgery.The osteoblasts and chondrocytes were isolated and cultured in vitro,and confirmed by alkaline phosphotase staining and toluidine blue staining.RT-PCR was performed to detect the neurofibromin mRNA,indirect immunofluorescence and Western blot were clone to confirm the expression of neurofibromin in osteoblasts and chondrocytes.Result:Type Ⅱ neurofibromin was expressed in these osteoblasts and cbondrocytes,tbe protein was mainly located in cytosol.Conclusion:Neurofibromin is expressed in osteoblasts and cbondrocytes from patients with congenital scoliosis,but the protein expressed is type Ⅱ isoform with relatively weak GAP activity.Whether neurofibromin contributes to the abnormalities of bone through its influence on osteoblasts and chondrocytes or not needs to be further studied.
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