淫羊藿苷对早期兔膝关节骨性关节炎的影响

Effect of Icariin on the osteoarthritic knee of rabbits in the early stage

【目的】研究淫羊藿苷(Icariin,ICA)对兔膝关节骨性关节炎早期影响。【方法】清洁级健康3月龄雄性日本大耳白兔18只。将动物随机分为对照组(Sham)、盐水组(ACLT+NS)和给药组(ACLT+ICA),每组各6只。ACLT+NS组和ACLT+ICA组行右膝关节前交叉韧带切断术(anterior cruciate ligament transaction,ACLT)造成膝骨性关节炎模型。术后ACLT+ICA组淫羊藿苷3mg.(kg.d)-1灌胃,每周称重1次,以调整药量,连续给药6周;ACLT+NS组则给予等剂量无菌生理盐水。术后6周处死动物,大体观察各组动物膝关节面情况,右侧股骨酒精固定之后脱钙切片,石蜡包埋行HE染色和MMP-13免疫组化染色,HE切片进行Mankin评分,MMP-13免疫组化染色切片用图像分析仪测量光密度;取右膝胫骨近端采取硬组织包埋方法,钨钢刀切片,用于骨组织形态计量学测量,观察骨小梁及软骨下骨。【结果】(1)ACLT+NS组较之Sham组与ACLT+ICA组关节软骨面呈显著破溃表现,关节周围有明显骨赘形成。(2)ACLT+NS组组织学Mankin评分结果显著高于Sham组与ACLT+ICA组(P<0.05)。(3)ACLT+NS组MMP-13免疫组化染色光密度值显著高于Sham组与ACLT+ICA组(P<0.05)。(4)ACLT+NS组软骨下骨骨小梁结构较之Sham组与ACLT+ICA组发生显著破坏(P<0.05)。【结论】淫羊藿苷能够下调MMP-13的表达,减轻兔膝关节软骨基质的降解,改善骨小梁结构,从而达到保护关节的作用。

【Objective】To investigate the effects of ICA on experimental osteoarthritic knee of rabbits induced by anterior cruciate ligament transaction surgery in early stage.【Methods】18 rabbits were randomly divided into three groups： Sham group（n = 6）,ACLT＋ NS group（n = 6） and ACLT＋ICA group（n = 6）.The rabbits in ACLT＋NS group and ACLT＋ICA group were received anterior cruciate ligament transaction（ACLT） operation at the right knee.The rabbits in ACLT＋ICA group were received a daily administration of ICA at a dose of 3 mg.（kg.d）-1 of body weight for 6 weeks,The other rabbits were received normal saline（NS） for 6 weeks.All the rabbits were sacrificed at six weeks after operation.The macro-pathologic changes of cartilage surface were observed,the sample of right femora were stained by HE and graded by Mankin scale.Expression level of MMP-13 were detected by immunstaining method.The subchondral bone of right tibia was prepared for analysis by bone histomorphometry.【Results】Compared with the rabbits in Sham group,the cartilage surface with significantly changes in ACLT＋NS group by macro-pathologic observation.The mankin score and the expression of MMP-13 in ACLT＋NS group was significantly higher than that in Sham group and in ACLT＋ICA group（P 0.05）.The micro-structure of subchondral trabecular were significantly destroyed in ACLT＋NS group as compared with that in in Sham group and in ACLT＋ICA group（P 0.05）.【Conclusion】ICA could inhibit the expression of MMP-13,so delayed the degradation of extra cell matrix.Otherwise,ICA exert partly protective effects on subchondral bone of OA knee in early stage.