补肾活血胶囊对激素性股骨头坏死大鼠骨修复和脂肪形成的影响

Effects of Bushen Huoxue Capsule on bone repair and adipogenesis in rats with steroid-induced osteonecrosis of femoral head

目的:观察补肾活血胶囊对激素性股骨头坏死(SONFH)大鼠骨修复与脂肪形成的影响,探讨其作用机制。方法:将30只雄性SD大鼠随机分为对照组,模型组,补肾活血胶囊高、中、低剂量组,每组6只,通过单纯注射甲强龙(MPS)构建SONFH大鼠模型,造模同时给予相应药物干预6周;干预结束后,取大鼠血清和双侧股骨头,进行血清生化学、Micro-CT、组织病理学检测,评估补肾活血胶囊对SONFH大鼠骨修复与脂肪形成的影响。结果:血清生化结果显示,补肾活血胶囊能降低血清过氧化物酶增殖剂激活受体γ(PPARγ)含量(P<0.05)。Micro-CT结果显示,模型组大鼠骨密度(BMD)较对照组显著降低(P<0.01),补肾活血胶囊低、中剂量组大鼠BMD显著高于模型组(P<0.01)。HE染色结果显示,模型组骨坏死明显,核收缩,股骨头骨小梁稀疏甚至消失,补肾活血胶囊各剂量组股骨头骨小梁组织结构病理改变明显减少;油红“O”染色结果显示,模型组大鼠软骨下骨局部区域脂肪组织明显增加,补肾活血胶囊各剂量组较模型组有改善;免疫组织化学染色结果显示,与模型组比较,补肾活血胶囊能增加大鼠局部骨小梁间隙存在中成骨相关因子Runt相关转录因子2(Runx2)、Ⅰ型胶原蛋白(CollagenⅠ)阳性蛋白表达水平、减少成脂相关因子重组人CCAAT增强子结合蛋白α(C/EBPα)、PPARγ蛋白表达水平。结论:补肾活血胶囊能有效地减少MPS引起的SONFH大鼠骨小梁丢失,抑制其脂肪形成,调节骨组织动态平衡而发挥治疗作用。

Objective: To observe the effects of Bushen Huoxue Capsule on bone repair and adipogenesis in steroidinduced necrosis of the femoral head(SONFH) rats, and to explore its mechanism. Methods: Thirty male SD rats were randomly divided into control group, model group, Bushen Huoxue Capsule high, medium and low dose groups, and 6 rats in each group.The model was given corresponding drug intervention for 6 weeks at the same time;after the intervention, the rat serum and bilateral femoral heads were collected for serum biochemical, Micro-CT, and histopathological tests to evaluate the effect of Bushen Huoxue Capsule on bone repair and adipogenesis in SONFH rats. Results: Serum biochemical results showed that Bushen Huoxue Capsule could reduce the content of serum peroxidase proliferator-activated receptor γ(PPARγ)(P<0.05). MicroCT results showed that the bone mineral density(BMD) of the rats in the model group was significantly lower than that in the control group(P<0.01), and the BMD of the rats in the low and medium dose Bushen Huoxue Capsule groups was significantly higher than that in the model group(P<0.01). Hematoxylin-Eosin staining results showed that the model group had obvious osteonecrosis, nuclear shrinkage, sparse or even disappearance of the femoral head trabecular bone, and the pathological changes of the femoral head bone trabecular tissue in each dose group of Bushen Huoxue Capsule were significantly reduced;The results of oil red ‘O’ staining showed that the adipose tissue in the subchondral bone of the model group increased significantly, and the groups of Bushen Huoxue Capsules were improved compared with the model group;The results of immunohistochemical staining showed that compared with the model group, Bushen Huoxue Capsule could increase the positive protein expressions of Runtrelated transcription factor 2(Runx2) and Collagen Ⅰ in the local trabecular bone space in rats, and could reduce the protein expression levels of adipogenic-related factors recombinant human CCAAT/enhancer binding protein(C/EBP) alpha(C/EBPα)and PPARγ. Conclusion: Bushen Huoxue Capsule could effectively reduce the loss of bone trabecular in SONFH rats induced by MPS, and inhibit its adipogenesis, and regulate the dynamic balance of bone tissue, and finally play a therapeutic role.