摘要
目的 通过观察去甲斑螯素对成骨细胞和破骨细胞的作用,探讨其在防治骨质疏松症中的作用.方法 将小鼠胚胎成骨细胞前体细胞分为对照组、模型组、低剂量组和高剂量组,高、低剂量组分别予以0.1 μmol/L、1 μmol/L 去甲斑蝥素干预.小鼠胚胎成骨细胞前体细胞用成骨培养基诱导,非对照组用脂多糖进行干预,3周后通过茜素红染色、碱性磷酸酶(ALP)染色检验成骨变化.骨髓来源巨噬细胞(BMMs)提取自6周龄的C57BL-6J小鼠,非对照组细胞培养基加入50 ng/ml RANKL向破骨细胞诱导6 d,通过抗酒石酸酸性磷酸酶(TRAP)染色、细胞骨架染色观察破骨细胞形态及活性.结果 ALP染色和茜素红染色显示,与模型组比较,去甲斑螯素干预组具有更强的碱性磷酸酶活性和更多的矿化结节,可有效改善脂多糖对成骨分化和矿化的抑制作用,增强成骨细胞活性.通过TRAP染色和细胞骨架染色观察,可见与模型组比较,去甲斑螯素干预后,成熟破骨细胞的数量和面积均显著减少,抑制了破骨细胞分化.结论 去甲斑螯素能通过剂量依赖性可有效增强成骨细胞活性,促进骨形成,并显著抑制破骨细胞生成,是一种治疗骨质疏松的理想药物.
Objective To investigate the effect of norcantharidin on the activity of osteoblast and osteoclast,and explore its role in the prevention of osteoporosis.Methods The experimental mice was divided into control group,model group,low dose group and high dose group.The high and low dose groups were intervened with 0.1μmol/L and 1μmol/L norcantharidin respectively.Murine pre-osteoblast MC3T3-E1 cells were induced by osteoblast differentiation medium,and other groups were intervened by lipopolysaccharide(LPS).After three weeks,alizarin red staining and Alkaline phosphatase(ALP)staining were employed to detect osteoblast differentiation.Bone marrow-derived macrophages(BMMs)were extracted from C57BL-6J 6-week-old mice.Other groups were added 50 ng/ml RANKL to induce osteoclasts for 6 days.The morphology and activity of osteoclasts were observed by TRAP staining and cytoskeleton staining.Results ALP staining and alizarin red staining showed that groups intervented by norcantharidin had higher alkaline phosphatase activity and more mineralized nodules than the model group.TRAP staining and Cytoskeletal staining showed that the number and area of mature osteoclasts were significantly reduced after the treatment with norcantharidin when comparing with the model group.Conclusion Norcantharidin can effectively enhance osteoblast activity,promote bone formation and significantly inhibit osteoclastogenesis in a dose-dependent manner.It is an ideal drug for the treatment of osteoporosis.
作者
黄飞日
姜文兵
苏忠良
HUANG Feiri;JIANG Wenbing;SU Zhongliang(Department of Orthopaedics,Wenzhou Poeple’s Hospital,Wenzhou 325000,China)
出处
《全科医学临床与教育》
2020年第5期407-410,F0002,F0003,共6页
Clinical Education of General Practice
