桃红四物汤对激素性股骨头坏死兔股骨头微结构的影响及其机制

摘要

目的 探讨桃红四物汤对激素性股骨头坏死兔股骨头微结构的影响及其可能的机制.方法 将50只健康成年新西兰大白兔随机分为观察组20只、模型组20只及正常对照组10只.模型组、观察组建立激素性股骨头坏死模型.建模后观察组给予桃红四物汤混悬液灌胃,模型组及正常对照组给予等体积超纯水灌胃,1次/d,连续12周.灌胃结束后各组进行高分辨率磁共振检查,计算冠状位坏死水肿区(ROE)面积、正常区(NR)面积及ROE/NR.各组处死后取出双侧股骨头,采用Micro-CT系统对左侧股骨进行扫描,记录相关参数:组织体积(TV)、骨体积(BV)、骨表面积(BS)、对连接密度(Conn.D.)、结构模型指数(SMI)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp),计算BV/TV、BS/BV.采用Western blotting法检测右侧股骨头组织成骨细胞特异性转录因子2(RUNX2)、骨桥蛋白(OPN)、过氧化物酶增殖物激活受体γ(PPARγ)、骨保护素(OPG)、多药耐药基因1(ABCB1)、核因子κB受体活化因子(RANK)、血管内皮生长因子(VEGF)、核因子κB受体活化因子配体(RANKL)蛋白相对表达量.结果 模型组ROE面积、ROE/NR均高于观察组及正常对照组,NR面积均低于观察组及正常对照组(P<0.05或<0.01).模型组BS/BV、Tb.Sp均高于观察组与正常对照组,Conn.D.、Tb.N均低于观察组与正常对照组(P<0.05或<0.01);模型组与观察组BV/TV均低于正常对照组,模型组Tb.Th、SMI均低于正常对照组(P均<0.05).模型组股骨头组织ABCB1、OPG、VEGF蛋白相对表达量均低于观察组及正常对照组,RANK、RANKL蛋白相对表达量均高于观察组及正常对照组(P均<0.05).模型组与观察组股骨头组织RUNX2蛋白相对表达量均高于正常对照组,且观察组升高更明显(P均<0.05).三组股骨头组织OPN、PPARγ蛋白相对表达量比较差异均无统计学意义 (P均>0.05).结论 桃红四物汤灌胃可提高激素性股骨头坏死兔股骨头的骨小梁数量及密度,改善其微结构状态,促进坏死组织修复;促进成骨、提高血管生长因子表达,抑制破骨、降低成脂因子表达可能是其作用机制.%Objective To observe the effect of Taohong Siwu decoction on the microstructure of femoral head in rabbits with steroid-induced osteonecrosis of the femoral head (SIONFH) and to explore its possible mechanism.Methods Fifty healthy New Zealand white rabbits were randomly divided into the model group (n=20), observation group (n=20), and normal control group (n=10).The SIONFH models in the model group and the observation group were established.After establishment, rabbits in the observation group were given the mixture of Taohong Siwu decoction by gavage.The rabbits in the model group and normal control group were given equal volume of normal saline, once a day, for 12 weeks.At the end of 12 weeks of gavage, high-resolution magnetic resonance (HRMR) was performed in each group to calculate coronal necrotic area (ROE), normal area (NR), and ROE/NR.The rabbits were sacrificed to obtain the bilateral femoral heads.The left femur was scanned by Micro-CT system.The parameters such as, tissue volume (TV), bone volume (BV), bone surface area (BS) connection density (Conn.D.), structural model index (SMI), number of trabecular bone (Tb.N), trabecular bone thickness (Tb.Th), trabecular bone separation (Tb.Sp) were recorded, and BV/TV and BS/BV were calculated.The protein expression of runt-related transcription factor 2 (RUNX2), osteopontin (OPN), peroxisome proliferator-activated receptor γ (PPARγ), osteoprotegerin (OPG), osteopontin (ABCB1), NF-κB receptor activating factor (RANK), vascular endothelial growth factor (VEGF), and nuclear factor κB receptor activator ligand (RANKL) in the right femur tissues were measured by Western blotting.Results The ROE and ROE/NR in the model group were higher than those in the observation group and normal control group, and NR was lower than that in the observation group and normal control group (P<0.05 or P<0.01).The levels of BS/BV and Tb.Sp in the model group were significantly higher than those in the observation group and the normal control group (P<0.05 or P<0.01).The levels of Conn.D.and Tb.N of the model group were lower than those in the observation group and the normal control group (P<0.05 or P<0.01).The level of BV/TV in the observation group and model group was lower than that in the normal control group, and the level of Tb.Th in the model group was lower than that of the normal control group (both P<0.05).The protein expression levels of ABCB1, OPG, and VEGF in the model group were lower than those in the observation group and the normal control group;the protein expression levels of RANK and RANKL in the model group were higher than those in the observation group and the normal control group (all P<0.05).The relative expression of RUNX2 protein in the femoral head tissues of the model group and the observation group was higher than that of the normal control group, and the observation group was more obvious (P<0.05).There was no significant difference in the relative protein expression of OPN and PPARγ between the three groups (both P>0.05).Conclusion Taohong Siwu decoction can improve the number and density of trabecular bone in femoral head of SIONFH rabbits, improve its microstructure, and promote the repair of necrotic tissues by promoting osteogenesis, increasing the expression of vascular growth factor, inhibiting osteoporosis, and decreasing the expression of lipid factor.