补肾壮筋汤对兔膝骨关节炎软骨形态学和骨架蛋白ROCK、Cofilin、Phospho-Cofilin、LIMK1和Phospho-LIMK1的影响

Influence of Bushen Zhuangjin Decoction on cartilage morphology and skeleton proteins of ROCK,Cofilin,Phospho-Cofilin,LIMK1 and Phospho-LIMK1 in rabbits with knee osteoarthritis

目的:探讨补肾壮筋汤对兔膝骨性关节炎(OA)模型形态学和细胞骨架蛋白的影响。方法:将48只3月龄新西兰大白兔随机分成4组,每组12只,分别为空白组、模型组、低剂量组和高剂量组,用Hulth法造成早期OA模型。空白组常规喂养;模型组以0.9%氯化钠溶液灌胃;低剂量组以补肾壮筋汤10.5g/kg灌胃,每天1次;高剂量组10.5g/kg灌胃补肾壮筋汤灌胃,每天2次,持续4周。采用HE染色观察软骨形态结构的变化,Western blot法检测OA软骨细胞ROCK、Cofilin、Phospho-Cofilin,LIMK1和Phospho-LIMK1蛋白含量变化。结果:HE:低剂量与高剂量组关节软骨退变程度较模型组轻(P<0.05);Western blot:软骨细胞骨架蛋白ROCK、Cofilin、PhosphoCofilin,LIMK1和Phospho-LIMK1蛋白表达,低剂量组与高剂量组均低于模型组(P<0.05)。结论:补肾壮筋汤可能通过调节细胞骨架信号通路相关蛋白的表达,抑制了软骨细胞骨架破坏的进程,减缓膝关节关节炎的退变。

Objective：To explore the influence of Bushen Zhuangjin Decoctioin on cartilage morphology and skeleton proteins in rabbits with knee osteoarthritis.Methods：Forty-eight 3 month old New Zealand rabbits were randomly divided into 4groups as control group,model group,low and high dosage of Bushen Zhuangjin Decoction groups,and with 12 rabbits in each group.The rabbit model of osteoarthritis in the early stage was established by using Hulth method.Rabbits in control group were fed conventionally,rabbits in model group received gavage administration with normal saline,rabbits inlow and high dosage of Bushen Zhuangjin Decoction groups received gavage administration with 10.5g/kg,once a day and Bushen Zhuanggu Decoction10.5g/kg,twice a day,and all the treatments were lasted for 4 weeks.The changes in shape and structure of cartilage were observed by using HE staining method,and the changes in the concentration of proteins as ROCK,Cofilin,Phospho-Cofilin,LIMKI and Phospho-LIMKI in cartilage were tested by using Western blot method.Results：The degeneration grade of articular cartilage of rabbits in low and high dosage of Bushen Zhuangjin Decoction groups was slighter than that of rabbits in model group（P〈0.05）.The expression of proteins as ROCK,Cofilin,Phospho-Cofilin,LIMKI and Phospho-LIMKI in rabbits of low and high dosage of Bushen Zhuangjin Decoction groups was lower than that of rabbits in model group,but which was higher than that of rabbits in control group（P〈0.05）.Conclusion：Bushen Zhuangjin Decoction could inhibit the development of cytoskeleton damage in cartilage and slower the articular degeneration by regulating the expression of related proteins of the cytoskeleton signal pathway.