摘要
目的 探究香叶木素调控Hedgehog信号通路对骨质疏松性骨缺损愈合的影响.方法 将SD大鼠分为假手术组、模型组、低剂量组(50 mg/kg香叶木素)、高剂量组(100 mg/kg香叶木素)及高剂量+环杷明组(100 mg/kg香叶木素+10 mg/kg Hedgehog信号通路特异性抑制剂环杷明),10只/组,切除双侧卵巢构建骨质疏松(osteoporosis,OP)大鼠模型,并在此基础上构建骨缺损模型,造模结束后进行相应灌胃给药;双能X线吸收扫描仪检测大鼠骨痂部位骨密度(bone mineral density,BMD);HE染色观察大鼠骨缺损区域病理学情况;抗酒石酸酸性磷酸酶(TRAP)染色观察大鼠骨缺损区域破骨细胞情况;qRT-PCR检测大鼠骨缺损区域组织成骨细胞特异性转录因子(Osterix)和Runt相关转录因子2(Runx2)表达;Western blot检测骨缺损区域组织音猬因子(SHH)、Gli家族锌指蛋白-1(GLI1)、跨膜蛋白受体1(Ptch 1)、Osterix和Runx2表达水平.结果 与假手术组相比,模型组大鼠骨痂部位BMD、骨小梁数、Osterix、Runx2、SHH、GLI1、Ptch1表达显著降低,TRAP阳性细胞数显著增加(P＜0.05);与模型组相比,低剂量组、高剂量组骨痂部位BMD、骨小梁数、Osterix、Runx2、SHH、GLI1、Ptch1表达显著增加,TRAP阳性细胞数显著降低(P＜0.05);环杷明可逆转香叶木素对骨质疏松性骨缺损愈合的影响.结论 香叶木素可通过激活Hedgehog通路促进骨质疏松性骨缺损愈合.
Abstract
Objective To explore the effect of diosmetin on the healing of osteoporotic bone defects by regulating Hedgehog signaling pathway.Methods SD rats were divided into sham operation group,model group,low-dose group(50 mg/kg diosmetin),high-dose group(100 mg/kg diosmetin),and high-dose+cyclopamine group(10 mg/kg cyclopamine,a Hedgehog signaling pathway specific inhibitor),with 10 rats in each group.Rat model of osteoporosis(OP)was built by removal of bilateral ovaries of the rat,and the bone defect model was built on this basis.After modeling,the corresponding drugs were given by gavage.Dual-energy X-ray absorptiometry was used to detect bone mineral density(BMD)in the callus of rats.HE staining was used to observe the pathology of the bone defect area in rats.Tartrate-resistant acid phosphatase(TRAP)staining was used to observe the osteoclasts in the bone defect area of rats.qRT-PCR was used to detect the expression of osteoblast-specific transcription factor(Osterix)and Runt-related transcription factor 2(RUNX2)in rat bone defect area.The expression levels of Sonic hedgehog(Shh),Gli family zinc finger-1(GLI1),Ptched 1(Ptch1),Osterix,and RUNX2 in the tissues of bone defect area were determined with Western blotting detection.Results Compared to those in the sham operation group,BMD at the callus,the number of trabeculae,the expressions of Osterix,Runx2,SHH,GLI1,and Ptch1 were significantly lower in the model group,while the number of TRAP positive cells was significantly higher(P＜0.05).Compared to those in the model group,BMD at the callus,the number of trabeculae,the expressions of Osterix,Runx2,SHH,GLI1,and Ptch1 were significantly higher in the low-dose group and high-dose group,while the number of TRAP positive cells was significantly lower(P＜0.05).Cyclopamine reversed the effect of diosmetin on the healing of osteoporotic bone defects.Conclusion Diosmetin promotes the healing of osteoporotic bone defects by activating the Hedgehog pathway.
维普
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