脱氢表雄酮对兔骨关节炎软骨ADAMTS/TIMP-3酶系统调节的实验研究  

Effect of dehydroepiandrosterone on the balance of ADAMTS/tissue inhibitor of metalloproteinases-3 system in rabbit osteoarthritis models

作　　者：黄恺[1] 蔡海丽[2] 吴立东[3] 徐卫星[1] 张春[1] 

机构地区：[1]浙江省立同德医院骨科,杭州310012 [2]解放军第117医院特诊科 [3]浙江大学医学院附属第二医院骨科

出　　处：《中华风湿病学杂志》2013年第11期764-768,F0003,共6页Chinese Journal of Rheumatology

基　　金：浙江省自然科学基金（Y2110078）;浙江省省级公益性技术应用研究计划项目（2013C37025）;浙江省医学会临床科研基金（2010ZYC-A06）

摘　　要：目的 观察关节腔内注射脱氢表雄酮（DHEA）对兔骨关节炎模型含血小板结合蛋白基序的解聚蛋白样金属蛋白酶（ADAMTS）/金属蛋白酶组织抑制因子（TIMP）-3酶系统的影响,并探讨其作用机制.方法 60只新西兰大白兔行双侧前交叉韧带切除术,术后4周,根据随机化原则于每只兔一侧膝关节腔内注射100 mmol/L DHEA（实验组）,另一侧关节腔内注射相同剂量的二甲基亚砜（对照组）,每周1次,共注射8周.术后12周行膝关节X线检查后处死兔子,取膝关节行大体和组织病理学检查,取股骨内髁软骨组织行羟脯氨酸及糖胺聚糖含量测定,用反转录聚合酶链反应方法检测关节软骨中蛋白聚糖酶-1、蛋白聚糖酶-2、TIMP-3、转化生长因子（TGF）-β1、蛋白聚糖、Ⅱ型胶原的表达,用蛋白印迹法检测关节软骨中Ⅱ型胶原及蛋白聚糖裂解片段（Aggrecan ARGxx）的表达.采用两样本Mann-Whitney检验或t检验进行统计学分析.结果 对照组股骨髁及胫骨平台软骨退变程度显著高于实验组（Z=-3.517,P＜0.01；Z=-2.518,P＜0.05）.实验组软骨病理学评分（5.3±1.2）显著低于对照组（10.1±1.3）,差异有统计学意义（P＜0.01）；实验组羟脯氨酸湿质量（5.7±0.3） μg/ml,干质量（23.6±1.7） μg/ml和糖胺聚糖[（30±4） μg/ml]含量显著高于对照组[湿质量（4.6±0.5） μg/ml,干质量（18.5±1.4）,糖胺聚糖（24±4）μg/ml],差异有统计学意义（P＜0.01）；实验组软骨ADAMTS-4 （0.15±0.03）、ADAMTS-5 （0.10±0.04）的mRNA表达显著低于对照组（0.29±0.08,0.15±0.05,P均＜0.05）；而TIMP-3 （0.85±0.10）、TGF-β1（1.2±0.4）、蛋白聚糖（0.87±0.31）及Ⅱ型胶原（2.7±0.6）的mRNA表达显著高于对照组（0.70±0.13,0.8±0.4,0.49±0.16,2.2±0.5）,差异有统计学意义（P均＜0.05）；实验组蛋白聚糖ARGxx（0.53±0.10）的蛋白表达显著低于对照组（0.81±0.12）；Ⅱ型胶原（2.3±0.7）蛋�Objective To study the effect of intra-articular injection of dehydroepiandrosterone （DHEA） on the balance of ADAMTS/TIMP-3 system in a rabbit osteoarthritis models.Methods Sixty rabbits were underwent bilateral anterior cruciate ligament transection （ACLT）.Rabbits were randomizedn to the following treatment：one knee of each rabbit was treated with 100 μmol/L DHEA dissolved in dimethylsulphoxide （the experimental group） and the other knee was treated under the same schedule using dimethylsulphoxide （the control group） 4 weeks after transection,once a week for eight weeks.Twelve weeks after ACLT,all rabbits were killed after X-ray assessment and the knee joints were evaluated by gross morphology and histology.The concentration of hydroxyproline and glycosaminoglycan in the cartilage were analyzed.The mRNA expression of ADAMTS-4,ADAMTS-5,tissue inhibitor of metalloproteinases-3 （TIMP-3）,transforming growth factor （TGF）-β1.Aggrecan and Collagen Ⅱ in the cartilage were analyzed using reverse transcription polymerase chain reaction （RT-PCR）.The protein expression of aggrecan ARGxx and Collagen Ⅱ in the cartilage were analyzed by Western blot.Results By Mann-Whitney test,Gross morphologic scores on femoral condyle and tibial plateau in the control group were significantly higher than the experi-mental group （Z=-3.517,P＜0.01 ; Z=-2.518,P＜0.05）.By unpaired Student＇s t test,histological evaluation showed that the grade of cartilage damage in the experimental group [（5.3±1.2） μg/ml] were less severe than that in the control group （10.1 ± 1.3,P＜0.01）.The concentration of hydroxyproline [（5.7±0.3,23.6± 1.7） μg/ml] and glycosaminoglycan （30±4） in the experimental group increased significantly when compared with the control group [（4.6±0.5）,（18.5±1.4）,（24±4） μg/ml,P＜0.01].The mRNA expression of ADAMTS-4 （0.15±0.03）and ADAMTS-5 （0.10±0.04） in the experimental group decreased significantly compared with the control group （0.29±0

关 键 词：骨关节炎 去氢表雄酮 金属蛋白酶3组织抑制剂 

分 类 号：R684.3[医药卫生—骨科学]