盐酸氨基葡萄糖联合降钙素对兔膝关节炎软骨和软骨下骨的保护作用

目的 研究盐酸氨基葡萄糖（GH）联合降钙素（CT）对兔骨性关节炎（OA）模型关节软骨和软骨下骨的保护作用及其作用机制.方法 将32只新西兰大白兔随机分为4组,每组8只,即假手术（Sham）组、右膝关节前交叉韧带切断术（ACLT）加0.9%氯化钠溶液（NS）治疗组、ACLT加GH治疗组、ACLT加GH加CT 联合治疗组.除Sham组外,其余24只动物右侧膝关节行ACLT.术后ACLT＋GH组动物给予GH150mg·kg-1·d-1灌胃;ACLT＋GH＋CT组动物给予GH灌胃,同时皮下注射CT 5 IU·kg-1·d-1;ACLT＋NS组动物给予等剂量NS灌胃及皮下注射.8周后处死所有动物,肉眼大体观察膝关节外观,取胫骨去除软骨后行软骨下骨生物力学检测（测定最大载荷和弹性模量）,取股骨测定远端1/4骨密度（BMD）后,脱钙、包埋、制备膝关节组织切片行HE染色及Mankin评分.结果 （1）大体观察：Sham组膝关节面软骨正常; ACLT＋NS组膝关节面发生明显退变;ACLT＋GH组、ACLT＋GH＋CT组膝关节亦有一定的OA表现,但程度较ACLT＋NS组明显减轻;（2）Mankin 评分：ACLT＋NS组显著高于Sham组,ACLT＋GH组、ACLT＋GH＋CT组显著低于ACLT＋NS组,但仍高于Sham组 （P＜0.05）;（3）股骨远端BMD：ACLT＋NS组显著低于Sham组,ACLT＋GH＋CT组最高,与ACLT＋NS组、ACLT＋GH组差别显著 （P＜0.05）;（4）生物力学检测结果：最大载荷、弹性模量ACLT＋NS组、ACLT＋GH组均显著低于Sham组（P＜0.05）,Sham组与ACLT＋GH＋CT组差别无统计学意义（P ＞0.05）.结论 GH与CT联合用药可抑制兔膝ACLT后OA的进程,效果优于单纯GH治疗,其作用机制可能为保护关节软骨及改善软骨下骨代谢的双重作用.

Protective effects of glucosamine hydrochloride combined with calcitonin on articular cartilage and subchondral bone of rabbit knee with anterior cruciate ligament transaction

Objective To investigate the protective effects and related mechanism of glucosamine hydrochloride combined with calcitonin on cartilage and subchondral bone in the osteoarthritic rabbit knees induced by anterior cruciate ligament transaction （ACLT）. Methods Thirty-two New Zealand white rabbits of 3m age were divided randomly into 4 groups （n=8 in each）. Rabbits in all groups except sham group received ACLT on the right knee. ACLT＋GH group received glucosamine hydrochloride （150mg.kg^-1.d^-1）,ACLT＋GH＋CT group received the same dose of GH plus a subcutaneous injection of CT （5 IU. kg^-1. d^-1）, ACLT＋NS group received normal saline only. All rabbits were sacrificed at week 8. Samples harvested from right knees were observed for the macro-pathologic changes and prepared for paraffin section and HE staining; the degeneration of cartilage was graded by Mankin＇s scale. The right tibias were removed for the biomechanical testa. Results Macro-pathologic changes： rabbits in sham group showed normal articular cartilage; osteoarthritic changes were observed in rabbits of ACLT＋NS group; compared with ACLT＋NS group, rabbits in ACLT＋GH group and ACLT＋GH＋CT group also presented but milder os- teoarthritic changes. Mankin＇s scores： rabbits of ACLT＋NS group showed significantly higher Mankin＇s scores than that of sham group （P〈0.05）, the scores in ACLT＋GH group and ACLT＋GH＋CT group decreased significantly compared with ACLT＋NS group （P〈0.05）, but was higher than that of sham rabbits （P〈0.05）. BMD： significantly lower BMD of the distal femurs were observed in ACLT＋NS rabbits compared to that of sham group（P〈0.05 ）, while rabbits in group ACLT＋GH＋CT had the highest BMD than those of the other three groups, but there was no significant difference compared to sham rabbits. Biomechanical tests： boththe maximal loading and compression modulus in ACLT＋NS and ACLT＋GH rabbits were significantly lower than those in sham group （P〈0.05）and ACLT＋GH＋CT group （P〈0.05）, but there was no significant difference between sham group and ACLT＋GH＋CT group. Conclusion GH combined with CT has better protective effects on ACLT-induced osteoarthritis in rabbits, which might be associated with the direct protection of cartilage and indirect effect on modulating subchondral bone metabolism.