通络生骨胶囊促进兔激素性股骨头坏死修复的机制研究

Mechanism of Tongluo Shenggu Capsules on Glucocorticoid-induced Osteonecrosis of Femoral Head in Rabbits

目的研究通络生骨胶囊促进新西兰兔激素性股骨头坏死修复的机制。方法 20只新西兰兔随机分为3组:正常组6只,模型组和中药组(通络生骨胶囊组)各7只,采用内毒素(LPS)联合甲泼尼龙琥珀酸钠(MPS)的方法制备激素性股骨头坏死模型,最后一次肌肉注射MPS后24 h,中药组给予0.3 g·kg-1通络生骨胶囊混悬液灌胃,每天一次,正常组和模型组灌服等体积超纯水,连续16周。灌胃16周后进行髋关节MRI检测和组织病理学检测以评估建模效果,采用实时荧光定量-聚合酶链反应(RT-q PCR)检测兔股骨头ABCB1、RUNX2、OPN、PPARγ的m RNA表达水平,采用Western Blot检测兔股骨头ABCB1、RUNX2、OPN、PPARγ的蛋白表达水平。结果与正常组比较,模型组的ABCB1和RUNX2的m RNA表达下调(P<0.05)、ABCB1的蛋白表达下调(P<0.05)、PPARγ的m RNA表达上调(P<0.05)。与模型组比较,中药组的ABCB1和RUNX2的m RNA和蛋白表达均上调(P<0.05,P<0.01),PPARγ的m RNA表达下调(P<0.05)。结论通络生骨胶囊促进新西兰兔激素性股骨头坏死的修复可能与调节ABCB1、RUNX2、PPARγ基因的功能有关。

Objective To investigate the mechanism of Tongluo Shenggu Capsules（TSC） on glucocorticoid- induced osteonecrosis of femoral head in rabbits. Methods Twenty New Zealand rabbits were randomly divided into three groups,normal control（6 rabbits）,model group（7 rabbits） and treatment group（7 rabbits）. Lipopolysaccharide（LPS）and methylprednisolone（MPS） were used together to induce the model of femoral head necrosis. The treatment group was given orally TSC 0.3 g·kg^（-1）,while the normal group and model group were given equal volume of normal saline through administration by gavage for 16 weeks 24 h after the last intramuscular injection of MPS. At the 16 th week of the experiment, MRI scaning of the hips and HE staining of femoral heads were performed for evaluating the osteonecrosis, while real- time quantitative PCR and Western blot assay were respectively applied for measuring the m RNA and protein expression of ABCB1,RUNX2,OPN and PPARγ in the bone tissues from femoral head. Results Compared to the normal group, m RNA expression of ABCB1 and RUNX2 and protein expression of ABCB1 were down- regulated（P〈0.05）, and m RNA expression of PPARγ was up- regulated in the model group（P〈0.05）.Compared to the model group,m RNA and protein expression levels of ABCB1 and RUNX2 were both up- regulated（P〈0.05, P〈0.01）, but m RNA expression of PPARγ was down- regulated in the treatment group（P〈0.05）.Conclusion The mechanism of TSC on promoting the recovery of glucocorticoid- induced osteonecrosis of femoral head may be related to the regulation of ABCB1,RUNX2,and PPARγ genes in rabbits.