四妙方对实验性骨关节炎大鼠软骨基质的影响

Effect of Simiao Prescription on Cartilage Matrix of an Eeperimental Osteoarthritis Rat Model

目的:观察四妙方对骨关节炎(osteoarthritis,OA)大鼠软骨基质的影响并探讨初步的作用机制。方法:健康雄性SD大鼠84只,随机分成假手术组、模型组、四妙方低、中、高剂量组、硫酸氨基葡萄糖组,每组14只。除假手术组外,其他组均采用大鼠前交叉韧带切断及部分半月板切除(anterior cruciate ligament transection and medial meniscus resection,ACLT+MMx)造成OA模型,造模后四妙方低、中、高剂量组分别给予四妙方0.63,1.25,2.50 g.kg-1.d-1治疗,硫酸氨基葡萄糖组ig给予硫酸氨基葡萄糖0.16 g.kg-1.d-1治疗,连续6周,假手术组和模型组分别给予等体积蒸馏水。6周后,腹主动脉采血测定Ⅱ型胶原C端肽(collagen typeⅡC-telopeptide,CTXⅡ)和Ⅱ型胶原C前肽(typeⅡprocollagen C-propeptide,CPⅡ);取大鼠右侧股骨进行HE染色观察四妙方对OA大鼠软骨病变的影响,甲苯胺蓝染色对软骨基质蛋白聚糖(proteoglycan,PG)的影响以及Ⅱ型胶原免疫组化染色对Ⅱ型胶原的影响。结果:与假手术组相比,模型组软骨表层变薄,软骨细胞明显减少,Mankin’s积分显著升高(P<0.01),甲苯胺蓝、Ⅱ型胶原免疫组化染色平均灰度值和Ⅱ型胶原阳性表达均显著降低(P<0.01),血清CTXⅡ显著升高(P<0.01),CPⅡ显著降低(P<0.01)。与模型组相比,四妙方1.25,2.50 g.kg-1均能显著降低Mankin’s积分(P<0.01),上调甲苯胺蓝染色平均灰度值(P<0.05,P<0.01),增加Ⅱ型胶原阳性表达量(P<0.01),下调CTXⅡ、上调CPⅡ水平(P<0.01)。结论:四妙方能有效减轻OA大鼠关节软骨基质破坏,其机制可能跟抑制软骨基质主要成分丢失、调节Ⅱ型胶原合成、降解标志物有关。

Objective： To investigate the effect of Simiao prescription on cartilage matrix of rat osteoarthritis, as well as to explore their mechanism of prevention and treatment for osteoarthritis（OA）. Method： Eighty-four male Sprague-Dawley rats were randomly divided into sham group,model group,Simiao prescription low,moderate and high does group,glucosamine sulfate group（n=14）. Except the sham group, underwent surgical destabilization （anterior cruciate ligament transection and partial medial meniscectomy） of the right knee to induce OA. The sham group and model group were fed with saline,glucosamine sulfate group were fed with glucosamine sulfate and others with Simiao prescription. After 6 weeks, using blood obtained from the abdominal aorta,and then measured the C-terminal peptide of collagen typeⅡand Collagen typeⅡC-propeptide, and then rat right femoral were removed.Sections of the articular cartilage were stained with HE, to observe cartilage damage, Toluidine blue stain, collagenⅡimmunohistochemistry stain was performed to assess the contents of proteoglycan and collagenⅡ. Result： Compared with the sham group,thinning of the cartilage surface of the model group, the cartilage cells decreased significantly and Mankin＇s score significantly higher, toluidine blue stain, collagen Ⅱ immunohistochemistry stain indicated that there was proteoglycan and collagen Ⅱ loss in the model group. The serum CTXⅡ was significantly increased,CPⅡ significantly decreased.Compared with model group, Simiao prescription can significantly reduce the Mankin＇s score, increase proteoglycan and type Ⅱ collagen content,lower levels of CTXⅡ raise CPⅡ. Conclusion： Simiao prescription can effectively reduce articular matrix destruction of experimental rat osteoarthritis, the metabolism may be related with inhibition of the main component of cartilage matrix, regulation of typeⅡcollagen synthesis and degradationmarkers.