一氧化氮合酶抑制剂对关节软骨修复的影响  

Repair effect of articular cartilage defects by nitric oxide synthase inhibitor

作　　者：王吉兴[1] 孙炜[1] 金大地[1] 刘晓霞 冯岚[1] 江建明[1] 

机构地区：[1]第一军医大学附属南方医院骨科,广州510515 [2]第一军医大学附属南方医院消化研究所病理研究室,广州510515

出　　处：《中华骨科杂志》2003年第2期108-113,共6页Chinese Journal of Orthopaedics

基　　金：国家自然科学基金资助项目(39900149)

摘　　要：目的观察一氧化氮合酶(iNOS)抑制剂S-甲基异硫脲(S-methylisothiourea,SMT)对关节软骨修复的影响,探讨NO在软骨修复过程中的作用。方法36只新西兰大白兔双侧股骨髁间关节面制成全层软骨缺损,随机分为3组:(1)对照组12只,软骨缺损区旷置;(2)rhBMP组12只,缺损区充填rhBMP纤维蛋白凝胶复合物;(3)SMT组12只,缺损区充填rhBMP纤维蛋白凝胶复合物后,皮下注射iNOS抑制剂SMT。术后16周处死动物,按组织形态学分级标准,作修复组织质量评价;天狼猩红-苦味酸染色观察修复组织内Ⅰ、Ⅱ型胶原分布;化学比色法检测NO释放量和NOS活性;35S掺入法检测蛋白多糖合成率;RT-PCR检测iNOS和MMP9mRNA的表达情况。结果术后16周,形态学评分表明,SMT组和rhBMP组在缺损区充填程度方面与对照组差异无显著性意义(F=2.32,P >0.05),在边缘修复结合度、细胞形态和缺损区结构以及软骨下骨修复等方面均优于对照组(P<0.05),其中SMT组优于rhBMP组(P<0.05)。化学比色法显示SMT组NO释放量(24.97±3.95)μmol/L和NOS活性(1.17±0.31)U/ml显著低于rhBMP组和对照组(F=21.287,F=15.932,P<0.05)。SMT组Ⅰ型胶原明显少于rhBMP组和对照组,Ⅱ型胶原多于rhBMP和对照组。SMT组35S掺入量(48276±5791.58)cpm明显高于对照组(10285±867.5)Objective To discuss repairing effects of articular cartilage defects by nitric oxide synthase inhibitor (S-methylisothiourea, SMT), and explore the role of nitric oxide in cartilage repair. Methods Full-thickness defects of cartilage were created in the intercondylar trochlear groove of femur of thirty-six adult New Zealand white rabbits, and were divided into three groups. Twenty-four defects were untreated as the control, twenty-four were filled with fibrin glue and impregnated with rhBMP as rhBMP group, the rest twenty-four were filled with fibrin glue and impregnated with rhBMP, and hypodermic injection with SMT as SMT group. The animals were sacrified at sixteen weeks postoperatively, and the gross appearance of the defect was estimated. The repair tissue was examined histologically and was evaluated according to the grading scale of histology. The amount of released NO and the activities of nitric oxide synthase (NOS) were examined by chemical colorimetry. The distribution of type-Ⅰ,Ⅱcollagen were examined by Sirius-Red. The proteoglycan synthesis was assessed by incorporation of radio-labelled sodium sulphate Na35SO42-. RT-PCR examined the expression of iNOSmRNA and MMP9mRNA. Results The filled extent of the defect in SMT group and rhBMP group had no significant difference from the control group, and the marginal integration, cellular morphology, architecture within the defect and subchondral plate repair were better than the untreated defects (P<0.05)according to the histological score. Sirius-Red stain demonstrated less significant type-Ⅰcollagen in the defects treated with rhBMP and SMT than in the untreated defects, and more type-Ⅱ collagen than in untreated defects. NO release (24.97±3.95) μmol/L and NOS activity (1.17±0.31) U/ml in SMT group were less than rhBMP and the control(F=21.287,F=15.932,P<0.05). Incorporation of isotope 35S was more significant in SMT (48 276±5 791.58)cpm than in rhBMP(37 624±5 217.34)cpm and the control (10 285±867.5)cpm (P< 0.05). MMP9mRNA expressed in all

关 键 词：一氧化氮合酶抑制剂 关节软骨 修复 INOS SMT 

分 类 号：R684[医药卫生—骨科学]