注射催产素对小鼠骨密度和成骨细胞分化影响的实验研究
Effects of oxytocin injection on bone mineral density and osteoblast differentiation in mice
  
DOI:10.3969/j.issn.1006-7108.2019.08.008
中文关键词:  催产素  骨代谢  骨密度  成骨细胞  骨质疏松
英文关键词:oxytocin  bone metabolism  bone mineral density  osteoblast  osteoporosis
基金项目:国家自然科学基金青年基金项目(81100625);教育部博士点基金(20110092120056)
作者单位
刘璇1*# 鲁荐1# 刘一鸣2 1 .东南大学医学院江苏 南京 210009 2 .南京市第一医院江苏 南京 210006 
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中文摘要:
      目的 通过建立雌性小鼠骨质疏松模型并给予催产素(oxytocin, OT)腹腔注射来验证催产素对骨代谢的调控作用。 方法 选取同批次2月龄雌性C57/B6小鼠15只,随机分成3组,每组5只: ①对照组(C),②造模组(OVX),③造模+催产素注射组(OVX+OT)。注射催产素前和注射8周后分别测定各组小鼠左股骨和腰椎L4-L6的骨密度,并提取小鼠骨髓细胞定向诱导分化,进行碱性磷酸酶(alkaline phosphatase, ALP)和Von Kossa染色以鉴定成骨细胞分化水平,通过qPCR检测分化相关因子表达水平。 结果 OVX组小鼠骨密度较C组显著降低(P<0.001),OVX+OT组小鼠骨密度较OVX组显著升高(P<0.001)。ALP染色显示,OVX组成骨细胞分化较C组差,细胞集落最少;OVX+OT组成骨细胞分化情况较OVX组好,染色面积增加了322.5%。Von Kossa染色发现OVX组细胞集落和钙结节较C组少且小,OVX+OT组细胞集落和钙结节较OVX组多,染色面积增加了62.3%。而进一步添加催产素体外培养的成骨细胞其分化水平均比未加入催产素组有所提高。qPCR结果显示催产素能够促进成骨相关基因OPN、Runx2和Osterix的表达(P<0.001)。 结论 催产素促进小鼠体内的骨合成代谢作用,提示其对骨质疏松症治疗具有潜在用途。
英文摘要:
      Objective To investigate and validate the regulation of oxytocin on bone metabolism by establishment of osteoporosis mouse model and injection of oxytocin in vivo. Methods Fifteen two-month-old C57/B6 mice were randomly divided into 3 groups, control group (C), OVX group (OVX), and OVX + oxytocin group (OVX+OT), with 5 mice in each group. After injecting oxytocin (50 μg/mice/day, s.c.) for 8 weeks, bone mineral density (BMD) of L4-L6 and the left femur of mice were measured. Mouse bone marrow cells were extracted and then stained with alkaline phosphatase (ALP) and von Kossa staining to identify the level of osteoblast differentiation. Moreover, qPCR was used to identify the expression levels of differentiation-related genes. Results BMD of mice in OVX group was markedly lower than that of mice in C group (P<0.0001). Oxytocin injection observably increased BMD of mice in OVX+OT group compared with mice in OVX group (P<0.0001). ALP staining results showed that the osteoblast differentiation is OVX group was worse than that in C group, and the cell colony was the least. The osteoblast differentiation in OVX+OT group was better than that in OVX group, with an increased dying area by 322.5%. von Kossa staining results showed that the cell colonies and calcium nodules in OVX group were smaller and less than that in C group. OVX+OT group had more cell colonies and calcium nodules compared with OVX group, with an increased dying area by 62.3%. Furthermore, the differentiation of osteoblasts with addition of oxytocin in vitro was better than that without oxytocin. qPCR results indicated that oxytocin up-regulated the expression of osteogenesis-related genes such as RUNX2, OSTERIX, and OPN at the mRNA level (P<0.0001). Conclusion Oxytocin enhances the anabolic action in regulating bone mass in mice, which suggests the potential use of this hormone for the treatment of osteoporosis.
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