康复新液对兔膝关节炎软骨缺损模型的修复作用及机制研究

Study on Repairing Effect of Kangfuxin Liquid on Cartilage Defect of Knee Osteoarthritis Model Rabbits and Its Mechanisms

目的:探讨康复新液对兔膝骨性关节炎(KOA)软骨缺损模型的修复作用及其机制。方法:取雄性新西兰兔72只,随机分为模型对照组和康复新低、中、高剂量组,每组18只。采用麻醉后手术钻孔的方式造成兔右膝关节股骨内侧髁软骨缺损模型,然后立即在软骨缺损处进行关节腔注射给药,康复新低、中、高剂量组分别给予体积分数为20%、40%、80%的康复新液,模型对照组给予等体积生理盐水,0.2 mL/kg,每3天1次持续给药。于给药后第4、8、12周时,观察兔软骨缺损伤口修复情况;于给药完毕即刻和给药后第4、8、12周时,结合Wakitani评分标准对兔软骨缺损处修复组织进行组织学评分;于给药后第12周,采用Masson染色法观察兔关节软骨缺损处修复组织的病理学变化;采用酶联免疫吸附法检测兔关节液中一氧化氮(NO)、超氧化物歧化酶(SOD)、过氧化脂质(LPO)和尿液中吡啶酚(PYD)水平。结果:给药后第4、8、12周时,与模型对照组比较,康复新低、中、高剂量组兔软骨缺损区修复情况较好;给药后第4、8、12周时,与给药完毕即刻及同时间点的模型对照组比较,康复新低、中、高剂量组兔膝关节软骨缺损处修复组织形态学评分均显著降低(P<0.05);给药后第12周时,与模型对照组比较,康复新低、中、高剂量组兔膝关节软骨缺损处组织病理学程度明显减轻;给药后第4、8、12周时,与模型对照组比较,康复新低、中、高剂量组兔关节液中NO、LPO和尿液中PYD水平均有不同程度地降低,SOD水平均有不同程度地升高,到给药第12周时,各指标差异均有统计学意义(P<0.05或P<0.01)。结论:康复新液对KOA软骨缺损模型兔的软骨损伤具有明显的修复作用,其作用机制主要与提高SOD表达并介导NO抑制软骨细胞凋亡有关。

OBJECTIVE: To investigate the effects of Kangfuxin liquid on repairing cartilage defect model of knee osteoarthritis(KOA)in rabbits and its mechanism. METHODS:Totally 72 male New Zealand rabbits were selected and randomly divided into model control group and Kangfuxin low-dose,medium-dose,high-dose groups,with 18 rabbits in each group. A cartilage defect model of the medial femoral condyle of the right knee joint in rabbits was established by drilling after anesthesia surgery. Then the rabbits in each group were given medicine via articular cavity immediately. Kangfuxin low-dose,middle-dose and high-dose groups were given 20%,40%,80% Kangfuxin liquid;model control group was given constant volume of normal saline consecutively,0.2 mL/kg,once every 3 days. At 4th,8th,12th week after medication,the wound repair of cartilage defect in rabbits was observed. Immediately after medication and at 4th,8th,12th week after medication,repaired tissue of cartilage defect in rabbits was scored histologically with Wakitani scoring standard under light microscope. At 12th week after medication,pathological changes of repaired tissue of cartilage defect in rabbits were observed by Masson staining. The levels of NO,SOD and LPO in joint fluid and PYD in urine of rabbits were detected by ELISA. RESULTS:At 4th,8th,12th week after medication,compared with model control group,cartilage defects in rabbits were repaired well in Kangfuxin low-dose,medium-dose and high-dose groups. At 4th,8th,12th week after medication,compared with immediately after medication and model control group at same time point,histomorphological score of repairing cartilage defect of knee joint in rabbits decreased significantly in Kangfuxin low-dose,medium-dose and high-dose groups(P<0.05). At 12th week after medication,compared with model control group,the histopathology degree of cartilage defect of knee joint in rabbits was significantly alleviated in Kangfuxin low-dose,medium-dose and high-dose groups. At 4th,8th,12th week after medication, compared with model control group, the levels of NO and LPO in joint fluid and PYD level in urine were decreased to different extent in Kangfuxin low-dose,medium-dose and high-dose groups, while SOD level was increased to different extent;at 12th week after medication,the difference of each index has statistical significance(P<0.05 or P<0.01). CONCLUSIONS: Kangangxin liquid can significantly repair cartilage defect of KOA cartilage defect model rabbits,the mechanism of which may be associated with increasing the expression of SOD and mediating NO-inhibited chondrocyte apoptosis.