补肾健脾活血方对大鼠悬尾试验性骨质疏松和肌萎缩影响的实验研究

Effect of Bushen Huoxue Jianpi Recipe on bone mineral density and muscle atrophy in tail suspended rats

目的:探讨补肾活血健脾方对悬尾大鼠骨密度和肌萎缩的影响。方法:采用SPF级健康SD大鼠48只,随机平均分为健脾活血方高剂量组,低剂量组,造模组和空白对照组,每组12只。除空白对照组外,其余各组均选用大鼠悬尾实验作为废用性骨质疏松和肌萎缩模型的建立动物模型造模共计28 d,造模期间,补肾健脾活血方高剂量组给予补肾健脾活血方提取物2.50 g/(kg·d)灌胃,补肾健脾活血方低剂量组给予补肾健脾活血方提取物1.25 g/(kg·d)灌胃,造模组和空白对照组给予等剂量饮用水灌胃,每天1次。28 d后,全部大鼠腹腔采血后处死,取右侧股骨以及L1~4椎骨和完整分离左右侧比目鱼肌。采用酶联免疫吸附法(ELISA)检测血清成骨标记物血清骨形成蛋白2(BMP-2)、骨钙素(OCN)及破骨标记物抗酒石酸酸性磷酸酶(TRAP)进行测定,应用实时荧光定量聚合酶链式反应(Real-time PCR)测定大鼠股骨成骨相关基因OCN,Runx-2的表达,双能X线检测各组椎体和股骨骨密度的表达,并测量比目鱼肌湿重与横截面积测量。结果:与造模组相比,健脾活血方高剂量组大鼠右侧股骨和椎体BMD显著性增高(P<0.05);健脾活血方高剂量和低剂量组血清TRAP呈显著性降低(P<0.01,P<0.05);健脾活血方高剂量和低剂量组大鼠股骨OCN和Runx-2 mRNA的表达显著性增高(P<0.05);健脾活血方高剂量组和低剂量组左侧和右侧比目鱼肌平均湿重显著性增高(P<0.05);健脾活血方高剂量组左侧和右侧比目鱼肌I型和II型纤维横截面积显著性增高(P<0.05),健脾活血方低剂量组左侧和右侧比目鱼肌I型纤维横截面积相比造模组呈显著性增高(P<0.05)。结论:补肾活血健脾方可以防止失重引起的骨密度降低和肌萎缩,为临床用于治疗废用性骨质疏松和肌萎缩奠定了理论基础。

Objective:To investigate the effect of Bushen Huoxue Jianpi Recipe on bone mineral density and muscle atrophy in tail suspended rats.Methods:48 healthy SD rats of SPF grade were divided into the high,low dose group of Jianpi Huoxue Formula,model group and blank group.The high dose group and the low dose group were given 2.50 g/(kg·d)and 1.25 g/(kg·d)of Bushen Jianpi Huoxue Decoction extract respectively,and the model group and the blank control group were given equal dose drinking water once a day.Rats in high and low dose group and placebo control group were suspended upside down for 28 days,while rats in non-tail suspension group were not suspended upside down.Rats in high and low dose of Bushen Huoxue Huayu recipe and water were intragastrically administered daily until the 28th day,respectively.The serum BMP-2 and TRAP markers were detected by ELISA.The expression of OCN and Runx-2 genes related to femoral osteogenesis was detected by real-time PCR.The expression of vertebral and femoral bone mineral density was detected by dual-energy X-ray.The wet weight and cross-sectional area of soleus muscle were measured.Results:Compared with the model group,the BMD of right femur and vertebral body in the high dose group of Jianpi Huoxue Formula was significantly higher(P<0.05);the serum trap of high dose and low dose group of Jianpi Huoxue Formula was significantly lower(P<0.01,P<0.05);the expression of OCN and Runx-2 mRNA of femur in the high dose and low dose group of Jianpi Huoxue Formula was significantly higher(P<0.05);The mean wet weight of left and right soleus muscle in the high dose and low dose groups of Jianpi Huoxue Formula increased significantly(P<0.05);The cross-sectional area of type I and type II soleus muscle fibers in the high dose group was significantly higher than that in the model group(P<0.05).Conclusion:Bushen Huoxue Jianpi Prescription can prevent the decrease of bone mineral density and muscle atrophy caused by weightlessness,which lays a theoretical foundation for clinical treatment of disused osteoporosis and muscle atrophy.