独活寄生汤对改良Hulth法造模大鼠膝骨关节炎软骨形态的影响

Effect of Duhuo Jisheng Decoction on Articular Cartilage Morphology of Rats with Knee Osteoarthritis on Modified Hulth Method

目的观察独活寄生汤对改良Hulth法造模大鼠膝骨关节炎软骨形态的影响,从形态学角度探讨独活寄生汤治疗膝骨关节炎的作用机制。方法2月龄SD大鼠66只,适应性喂养1周后,随机分为正常组(18只)和造模组(48只)。应用改良Hulth法复制大鼠膝骨关节炎模型,造模2周后,从正常组和造模组中随机抽取3只大鼠进行小动物MRI观察,鉴定造模是否成功。造模成功后,采用抽签法分为模型组、对照组和治疗组各15只。正常组和模型组给予0.9%生理盐水,治疗组给予独活寄生汤,对照组给予盐酸氨基葡萄糖胶囊灌胃。治疗12周后,切取左侧胫骨平台内侧软骨组织,固定、脱钙、石蜡包埋,行HE染色与番红O-固绿染色,并分别采用Mankin软骨组织学评分、OARSI关节病理评分评价染色结果。结果MRI显示:正常组可见关节间隙正常,关节面平整、光滑;造模组关节间隙增宽,关节软骨面不平整、软骨层变薄、局部缺损,结缔组织增生,骨赘形成,关节肿胀积液明显。与正常组比较,模型组软骨形态病变严重,Mankin评分、OARSI评分显著增高(P<0.01);治疗组与对照组软骨形态病变较轻;与对照组比较,治疗组软骨形态病变较轻,Mankin评分、OARSI评分明显降低(P<0.01)。结论改良Hulth法可成功复制大鼠膝骨关节炎模型。独活寄生汤能延缓软骨退变,促进软骨生长与修复,保护膝骨关节炎大鼠软骨形态。

Objective: To explore treatment mechanism of Duhuo Jisheng decoction(DHJSD) from morphology point by observing its effects on articular cartilage morphology of rats with knee osteoarthritis on modified Hulth method. Methods: After one week of acclimation, two-month-old male SD rats(n=66) were randomly divided into normal group(n=18) and osteoarthritis model group(n=48). The model group was made according to modified Hulth method, and penicillin was injected on time to prevent infection.Two weeks after operation, magnetic resonance imaging(MRI) of the knee joint was taken for identification. After successfully establish the model, the left rats were randomly divided into three groups, including model group(n=15), control group(n=15) and treatment group(n=15). Normal group and model group were given 0.9% NaCl solution, control group was given glucosamine hydrochloride capsule, and the treatment group was given DHJSD for 12 weeks via gavage. The medial tibial plateau cartilage tissue of left knee was removed and fixed in 4% paraformaldehyde after the treatment. The specimens were embedded in paraffin after decalcification for 8 weeks. HE stain and Safranin O-fast green staining were performed to observe histological changes of knee cartilage and evaluated according to Mankin score and OARSI grade. Results: MRI showed that in normal group, the articular space was normal and the articular surface was even and smooth. Compared with the normal group, the articular cartilage of the model group was thinner with local filling defect, and the intra-articular effusion increased significantly. The cartilage morphology of the model group was severe, and the Mankin score and OARSI score were significantly increased compared with the normal group. The control group and the treatment group had milder cartilage morphology. Among them, the treatment group was lighter than the control group, and Mankin score and OARSI score is lower. Conclusion: The modified Hulth method could successfully replicate the rat knee osteoarthritis model. DHJSD can ameliorate the pathological changes of knee articular cartilage in rats with knee osteoarthritis, so as to delay cartilage degeneration and protect the cartilage.