补肾化痰祛瘀方对铜负荷Wilson病模型TX小鼠骨代谢异常的影响

Effect of Kidney-tonifying Phlegm-Dispelling Stasis formula on bone metabolism abnormality in Copper-loaded Wilson’s Disease model TX mice

目的探讨补肾化痰祛瘀方对Wilson病TX小鼠骨代谢异常的影响及其作用机制。方法TX和正常对照小鼠随机分为空白组(Blank group)、Wilson病模型组(Wilson disease model group)、二巯丁二酸组(DMSA group)、补肾化痰祛瘀方(BSHTQYF group)、补肾化痰祛瘀方+DMSA组(BSHTQYF+DMSA),每组10只。运用苏木精-伊红染色法观察小鼠股骨组织病理形态结构变化,番红染色法进一步辨析小鼠软骨组织的剥脱与生长;免疫组化法检测小鼠股骨组织中骨钙蛋白(Osteocalcin,BPG)与骨保护蛋白(Osteoprotegerin,OPG)的表达情况,并测定其平均光密度水平。结果与空白组相比,Wilson病模型组小鼠股骨组织形态结构紊乱变形;股骨组织中的BPG蛋白及OPG蛋白阳性表达及平均光密度水平显著下降(P<0.01);补肾化痰祛瘀方组上述指标较Wilson病模型组改善明显(P<0.01)。结论补肾化痰祛瘀方能促进TX小鼠股骨组织的生长发育,提高TX小鼠骨组织中BPG蛋白及OPG蛋白表达水平,改善TX小鼠的骨代谢障碍情况。

Objective To investigate the effect of tonifying kidney,removing phlegm and removing blood stasis on abnormal bone metabolism in Wilson’s Disease TX mice and its mechanism.Methods in SPF grade laboratory environment,50 TX mice were fed for 14 days,and randomly divided into blank group,Wilson disease model group,dimermeric butanic acid group(DMSA group),Tonifying kidney and resolving phlegm and eliminating stasis formula group(BSHTQYF group),and Tonifying kidney,removing phlegm and removing blood stasis+DMSA group(BSHTQYF+DMSA),with 10 mice in each group.Hematoxylin-Eosin staining was used to observe the pathological morphological changes of femur tissues in mice,and saffron staining was used to further differentiate and analyze the exfoliation and growth of mouse cartilage tissues.Immunohistochemical method was used to detect the expression of Osteocalcin(BPG)and Osteoprotegerin(OPG)in mouse femur tissues,and the average optical density level was determined.Results Compared with the blank group,the morphological structure of femur in Wilson disease model group was disordered and deformed.The positive expression of BPG and OPG proteins in femur tissues and the average optical density level decreased significantly(P<0.01);The above indexes in the group of BSHTQYF were significantly improved compared with those in the Wilson disease model group(P<0.01).Conclusion BSHTQYF can promote the growth and development of femur tissues in TX mice,improve the expression levels of BPG and OPG proteins in bone tissues of TX mice,and improve the bone metabolism disorder in TX mice.