桃红四物汤对创伤性股骨头坏死模型大鼠骨组织BMP-4和Osterix表达的影响

Effect of Taohong Siwu Decoction on the expressions of BMP-4 and Osterix in bone tissue of model rats with trauma-induced osteonecrosis of the femoral head

目的:通过检测桃红四物汤对创伤性股骨头坏死(trauma-induced osteonecrosis of the femoral head,TIONFH)模型大鼠骨组织BMP-4和Osterix表达的影响,探讨桃红四物汤延缓TIONFH病程的作用机制。方法:将35只SD大鼠适应性喂养1周后随机分为空白组(7只)和造模组(28只)。造模组按照经典股骨头血供剥离法进行造模,造模4周后随机抽取空白组1只、造模组4只进行对比,确认造模成功后,将造模组随机分为桃红四物汤高、中、低剂量组及模型组,共4组,每组6只。依据人与动物体表面积换算公式,桃红四物汤组分别灌服相应剂量的汤剂,另外两组灌服等量0.9%氯化钠注射液,每日2次,持续6周。干预6周后取材,观察并记录股骨头大体形态,进行骨组织HE染色,采用Real-time-PCR法和Western Blot法分别检测股骨头组织中BMP-4和Osterix的基因和蛋白表达情况。结果:经桃红四物汤干预后,桃红四物汤高、中剂量组股骨头病理情况优于模型组,次于空白组。Western Blot结果显示,与空白组比较,模型组大鼠股骨头组织BMP-4、Osterix蛋白表达水平明显降低(P<0.01);与模型组比较,桃红四物汤各剂量组BMP-4蛋白表达水平明显升高(P<0.05,P<0.01),桃红四物汤高、中剂量组Osterix蛋白表达水平明显升高(P<0.01),桃红四物汤低剂量组Osterix蛋白表达水平与模型组比较,差异无统计学意义(P>0.05)。Real-time-PCR结果显示,与空白组比较,模型组大鼠股骨头组织BMP-4、Osterix mRNA表达量明显降低(P<0.01);与模型组比较,桃红四物汤高、中剂量组BMP-4、Osterix mRNA表达量明显升高(P<0.01);桃红四物汤低剂量组BMP-4、Osterix mRNA表达与模型组比较,差异无统计学意义(P>0.05)。结论:桃红四物汤能改善股骨头病理形态,其机制可能与桃红四物汤提高成骨相关因子BMP-4、Osterix的表达从而促进骨修复有关。

Objective:To investigate the effect of Taohong Siwu Decoction on the expression of BMP-4 and Osterix in bone tissue of model rats with trauma-induced osteonecrosis of the femoral head(TIONFH)and explore the mechanism of Taohong Siwu Decoction in delaying the course of TIONFH.Methods:35 Sprague-dawley rats were randomly divided into blank group(n=7)and modeling group(n=28)after 1 week of adaptive feeding.The modeling group was modeled ac-cording to the classic femoral head vascular deprivation method.And after 4 weeks of modeling,1 rat in the blank group and 4 rats in the modeling group were randomly selected to confirm the success of modeling.Then the modeling group was ran-domly divided into high,medium and low dose groups of Taohong Siwu Decoction and model group.There were 4 groups with 6 animals in each group.According to the conversion formula of human and animal body surface area,the Taohong Siwu Decoction group were given corresponding decoction respectively,and the other two groups were given the same amount of 0.9%sodium chloride injection.Twice daily for 6 weeks.After 6 weeks of intervention,the samples were taken to observe the morphology of femoral head,and the HE staining was performed on the bone tissue.Real-time PCR and Western Blot were used to detect the gene and protein expressions of BMP-4 and Osterix in the femoral head tissue,respectively.Results:After the intervention of Taohong Siwu Decoction,the pathological condition of femoral head in high-dose and medium-dose groups was superior to model group and inferior to blank group.Western Blot results showed that compared with blank group,the protein expression levels of BMP-4 and Osterix in femoral head tissue of rats in model group were significantly decreased(P<0.01).Compared with model group,the expression of BMP-4 protein in each dose group of Taohong Siwu Decoction was significantly increased(P<0.05 or P<0.01).The expression level of Osterix protein in high-dose and medium-dose groups of Taohong Siwu Decoction was significantly increased(P<0.01),and there was no statistical significance in the expression level of Osterix protein between the Taohong Siwu Decoction low-dose group and the model group(P>0.05).Real-time PCR results showed that compared with blank group,BMP-4,Osterix and mRNA in femoral head tissue of rats in model group were signif-icantly decreased(P<0.01).Compared with model group,BMP-4,Osterix and mRNA in high-dose and medium-dose groups of Taohong Siwu decoction were significantly increased(P<0.01).The BMP-4,Osterix and mRNA in Taohong Siwu decoction low-dose group were not statistically different from those in model group(P>0.05).Conclusion:Taohong Siwu Decoction can improve the pathomorphology of femoral head and exert its therapeutic effect,which may be related to the enhancement of the expression of osteogenic factors BMP-4 and Osterix by Taohong Siwu Decoction,thus promoting bone repair.