秦皮乙素促进成骨细胞分化延缓增龄性骨质疏松的研究
Esculetin enhances osteogenesis and prevents senile osteoporosis
  
DOI:10.3969/j.issn.1006.7108.2018.07.004
中文关键词:  中医中药  秦皮乙素  骨质疏松  成骨细胞分化  骨密度
英文关键词:Chinese Traditional Medicine  Esculetin  Osteoporosis  Osteoblast differentiation  Bone mineral density
基金项目:浙江省医药卫生科技计划(2018KY194,2014RCA001);浙江省中医药科技项目(2018ZA003)
作者单位
王继荣 暴一众 杨舟鑫 吕晓玲 胡细连 王国付* 浙江医院浙江省老年医学重点实验室浙江 杭州 310013 
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中文摘要:
      目的 探讨秦皮乙素对成骨细胞分化的调控及对增龄性骨质疏松症的治疗作用。方法 取7月龄C57BL/6小鼠随机分为对照组、秦皮乙素200 mg/kg组与秦皮乙素400 mg/kg组,分别灌胃给予生理盐水与秦皮乙素。给药3个月后取材,X射线检测小鼠胫骨骨密度,qPCR检测成骨细胞分化标记基因表达,成骨诱导液诱导28 d后茜素红染色检测成骨细胞矿化结节形成能力,胫骨组织HE染色观察骨组织形态。结果 10月龄时灌胃给予200 mg/kg秦皮乙素的小鼠胫骨骨密度略高于对照组,而灌胃给予400 mg/kg秦皮乙素的小鼠胫骨骨密度明显高于对照组。HE染色结果同样显示400 mg/kg组小鼠骨小梁显著多于对照组。成骨诱导分化与茜素红染色结果显示,秦皮乙素400 mg/kg组骨髓来源间充质干细胞矿化结节形成能力增强。qPCR结果表明秦皮乙素剂量依赖地促进成骨细胞分化标记基因碱性磷酸酶、骨涎蛋白、Col1与Runx2的表达,同时发现经典Wnt信号关键基因Dkk1与Lef1表达也明显升高。结论 秦皮乙素促进成骨细胞分化能力,进一步延缓增龄性骨质疏松症,其可能与调控经典Wnt信号相关。
英文摘要:
      Objective To investigate the effects of esculetin on osteoblasts differentiation and its therapeutic effects on senile osteoporosis. Methods 7-month-old C57BL/6 mice were assigned randomly into control group, esculetin 200 mg/kg and esculetin 400 mg/kg groups, and were intragastric administrated with saline (control group) or esculetin daily at 200 or 400 mg/kg for 3 months, respectively. After mice were sacrificed, analysis of the bone mineral density (BMD) was performed by X-ray. The expression of osteoblast speci?c genes were assayed by qPCR in BMSCs cells. After incubation for 28 days, cells were detected for bone nodules by alizarin-red staining and quantitative determination. The morphology of bone was tested by HE staining of paraffin sections of the proximal tibia from mice. Results X-ray of the proximal tibia confirmed that esculetin at both 200 and 400 mg/kg resulted in increase in bone mineral density. The results of HE staining showed that the number of trabecular bone increased significantly in the 400 mg/kg group. Moreover, esculetin not only increased mRNA levels of osteoblastogenic markers including alkaline phosphatase (ALP), bone sialoprotein (BSP), type I collagen (Col1), and Runt-related transcription factor 2 (Runx2), but also increased canonical Wnt signaling target genes including lymphoid enhancer-bindingfactor-1 (Lef1) and Dickkopf-1(Dkk1), as well as mineralized nodule formation. Conclusion Esculetin promoted osteogenesis and resisted bone loss in senile osteoporosis through Wnt signaling cascades.
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