血管细胞黏附分子-1在大骨节病软骨细胞异常分化过程中的作用  

Role of vascular cell adhesion molecule-1 in abnormal differentiation of chondrocytes in Kaschin-Beck disease

作　　者：邵明明 张萌[1] 张迎 张丹[1] 张莹[1] 刘慧中[1] 何颖[1] 王梦莹[1] 曲梦[1] 卢洁 孙健 陈静宏 Shao Mingming;Zhang Meng;Zhang Ying;Zhang Dan(Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Institute of Endemic Diseases, School of Medicine, Xi＇an Jiaotong University, Xi＇an 710061, China;Medical Laboratory Center, Department of Medical Technology, Xi＇an Medical University, Xi＇an 710021, China;College of Genetics and Molecular Biology, School of Medicine, Xi＇an Jiaotong University, Xi＇an 710061, Chin)

机构地区：[1]西安交通大学医学部地方病研究所国家卫生计生委微量元素与地方病重点实验室,西安710061 [2]西安医学院医学技术系医学检验实验中心,西安710021 [3]西安交通大学医学部遗传学与分子生物学系,西安710061

出　　处：《中华地方病学杂志》2018年第7期547-553,共7页Chinese Journal of Endemiology

基　　金：国家自然科学基金重点项目（81573102、81273006）

摘　　要：目的观察血管细胞黏附分子-1（vascular cell adhesion molecule-1，VCAM-1）在氧化应激诱导的肥大软骨细胞、大骨节病（KBD）儿童和成人关节软骨以及低硒和T-2毒素中毒大鼠膝关节软骨的表达变化，探讨VCAM-1在大骨节病深层软骨细胞坏死以及分化异常中的作用。方法采用1％混合液（含10mg/L胰岛素、5．5mg／L转铁蛋白、6．7μg/L亚硒酸钠）对小鼠软骨前体细胞ATDC5作用21d，诱导为肥大软骨细胞，选择5-氨基-3-（4-吗啉基）-1，2，3-恶二唑盐酸盐（3-morpholino-sydnonimine hydroehloride，SIN-1）作为自由基供体构建氧化应激致肥大软骨细胞坏死模型。采用荧光定量（Real-time）PCR检测不同浓度SIN-1诱导的肥大软骨细胞中VCAM-1mRNA表达情况。采用免疫组织化学技术检测KBD儿童、成人及对照组关节软骨各层软骨细胞VCAM-1表达情况。采用免疫组织化学技术检测低硒和T-2毒素中毒KBD大鼠动物模型膝关节软骨及骺板软骨各层软骨细胞VCAM-1的表达水平。结果随SIN-1浓度梯度（0、1、3、5mmoL／L）增高，诱导的小鼠关节软骨肥大细胞中VCAM-1mRNA表达降低（1．00±0．00、1．22±0．20、0．71±0．22、0．37±0．16），组间比较差异有统计学意义（F=27．788，P〈0．05）；KBD儿童关节软骨表层、中层VCAM-1阳性细胞率[（16．08±5．20）％、（19．20±9．71）％]高于对照组[（0．00±0．00）％、（0．00±0．00）％]，深层VCAM—1阳性细胞率[（7．00±4．40）％]低于对照组[（51．60±20．58）％，t表、中、深=-10．972、-6．249、6．564，P均〈0．05]。KBD成人关节软骨表层VCAM-1阳性表达率[（7．92±4．29）％]高于对照组[（3．12±1．12）％]，中层[（17．54±8．27）％]表达低于对照组[（31．75±13．30）％]，组间比较差异有统计学意义（t表、中=-3．824、3．037，P〈0．05）。动物实验中，与对照组[（1．89±1．76）％]比较。低硒Objective To investigate the expression of vascular cell adhesion molecule-1 （VC.AM-1） in oxidative stress induced hypertrophic chondrocytes, in Kaschin-Beck disease （KBD） patients and in rat fed with T-2 toxin under selenium deficient conditions in order to analyze the relationship between VCAM-1 biological function and the dysregulation of chondrocyte differentiation in KBD. Methods The ATDC5 was cultured in 1% ITS solution （10 mg/L insulin, 5.5 mg/L transferrin, and 6.7 μg/L sodium selenite） for 21 days, and stimulated with 3- morpholino--sydnonimine （SIN-1, a nitric oxide [NO] donor） to obtain the oxidative stress induced hypertrophic chondrocytes. Real-time PCR was used to detect VCAM-1 mRNA in hypertrophic chondrocytes induced by different concentrations of SIN-1. The expressions of VCAM-1 in articular cartilage of child and adult KBD patients and KBD animal model were determined via the immunohistochemical method, and KBD cartilage samples were obtained in KBD areas from KBD child who had died or from adults who had had surgery. Results After treatment of hypertrophic chondrocytes （ATCD5 cells） with SIN-1 （0, 1, 3, 5 mmol/L）, VCAM-1 mRNA levels （1.00 ± 0.00, 1.22 ± 0.20, 0.71 ± 0.22, 0.37 ± 0.16） were decreased in a dose-dependent manner when compared with the control group （F = 27.788, P 〈 0.05）. The densities of VCAM-1 positive ceils in superficial and middle zones of the articular cartilage of children KBD patients [（16.08 ± 5.20）%, （19.20 ± 9.71）%] were higher than those of control group [（0.00 ± 0.00）%, （0.00 ± 0.00）%], while that in the deep zone [（7.00 ± 4.40）%] in children KBD patients was significantly lower than that of control [（51.60 ± 20.58）%,tS/M/D=- 10.972, - 6.249, 6.564, P 〈 0.05]. The positive cell density of VCAM-1 in the adult patients was significantly increased in the superficial zone [（7.92 ± 4.29）% vs （3.12 ± 1.12）%] but significantly decreased in the middle zone [（17.54 ± 8.27）% vs （31.75 ± 13.

关 键 词：大骨节病 血管细胞黏附分子-1 氧化应激 分化障碍 

分 类 号：R684.1[医药卫生—骨科学]