加味补阳还五汤对兔膝骨关节炎的治疗作用及机制

Therapeutic effect and mechanism of modified Buyang Huanwu decoction on knee osteoarthritis in rabbits

目的观察加味补阳还五汤灌胃对兔膝骨关节炎的治疗作用,并探讨其作用机制。方法将24只健康新西兰兔随机分为正常组、模型组、壮骨关节丸组、加味补阳还五汤组各6只。除正常组外,其他各组采用改良Hulth关节内手术法+驱赶法制备兔膝骨关节炎模型。于造模第2周开始,壮骨关节丸组给予壮骨关节丸按1.2 g/(kg·d)灌胃,加味补阳还五汤组给予加味补阳还五汤18.4 g/(kg·d)灌胃,模型组给予相应体积蒸馏水灌胃。各组均连续灌胃8周。麻醉过量处死动物,取其左侧胫骨平台及股骨关节面,保留软骨,观察关节腔表面关节软骨形态。HE染色后观察膝关节组织病理变化,用TUNEL法检测膝关节软骨细胞凋亡率,用免疫组化法检测膝关节软骨细胞Bcl-2、Bax蛋白表达。结果模型组膝关节呈炎症性改变,可见明显关节积液,关节软骨表面粗糙不平,关节滑膜增生,局部可见骨赘形成,光学显微镜下可见软骨细胞排列紊乱,有细胞簇集现象;加味补阳还五汤组、壮骨关节丸组关节软骨损伤、组织病理变化均明显轻于模型组。与正常组比较,模型组细胞凋亡率高(P<0.05);与模型组比较,壮骨关节丸组、加味补阳还五汤组细胞凋亡率低(P均<0.05);壮骨关节丸组与加味补阳还五汤组细胞凋亡率比较差异无统计学意义(P>0.05)。与正常组比较,模型组膝关节软骨细胞中Bcl-2蛋白表达低,Bax蛋白表达高(P均<0.05);与模型组比较,壮骨关节丸组、加味补阳还五汤膝关节软骨细胞中Bcl-2蛋白表达高,Bax蛋白表达低(P均<0.05);壮骨关节丸组与加味补阳还五汤组膝关节软骨组织中Bcl-2、Bax蛋白表达比较差异无统计学意义(P均>0.05)。结论加味补阳还五汤灌胃对兔膝骨关节炎有治疗作用;其作用机制可能通过调控Bcl-2、Bax蛋白表达抑制软骨细胞凋亡。

Objective To observe the therapeutic effect of intragastric administration of modified Buyang Huanwu decoction on rabbit knee osteoarthritis and to explore its mechanism.Methods Twenty-four healthy New Zealand rabbits were randomly divided into the normal group(n=6),model group(n=6),Zhuanggu joint pill group(n=6)and modified Buyang Huanwu decoction group(n=6).Except for the normal group,the rabbit knee osteoarthritis model was established by modified Hulth intra-articular operation and expelling method.From the second week of modeling,rabbits in the Zhuanggu joint pill group were given Zhuanggu joint pill at 1.2 g/(kg·d),the rabbits in the modified Buyang Huanwu decoction group were given modified Buyang Huanwu decoction 18.4 g/(kg·d),and the rabbits in model group were given corresponding volume of distilled water.All groups were intervened continuously for 8 weeks.The animals were killed under excessive anesthesia.The left tibial plateau and femoral articular surface were taken to retain cartilage,and the morphology of articular cartilage on the surface of articular cavity was observed.After HE staining,the pathological changes of knee joint tissues were observed,the apoptosis rate of knee joint chondrocytes was detected by TUNEL method,and the expression of Bcl-2 and Bax protein in the knee joint chondrocytes was detected by immunohistochemical method.Results The knee joint in the model group showed inflammatory changes,with obvious articular effusion,rough surface of articular cartilage,synovial hyperplasia and local osteophyte formation.The articular cartilage injury and histopathological changes in the modified Buyang Huanwu decoction group and Zhuanggu Joint Pill group were significantly slighter than those in the model group.Compared with the normal group,the apoptosis rate in the model group was higher(P<0.05),and that in the Zhuanggu joint pill group and modified Buyang Huanwu decoction group was lower than that in the model group(P<0.05).There was no significant difference in apoptosis rate between Zhuanggu joint pill group and Jiawei Buyang Huanwu decoction group(P>0.05).Compared with the normal group,the expression of Bcl-2 protein was lower and the expression of Bax protein was higher in the model group;the expression of Bcl-2 protein was higher and the expression of Bax protein was lower in the Zhuanggu joint pill group and modified Buyang Huanwu decoction group as compared with those of the model group.There was no significant difference in the expression of Bcl-2 or Bax protein between Zhuanggu joint pill group and modified Buyang Huanwu decoction group.Conclusion Modified Buyang Huanwuyang decoction through intragastric administration has a therapeutic effect on rabbit knee osteoarthritis by regulating the expression of Bcl-2 protein and Bax protein to inhibit the apoptosis of chondrocytes.