申晋斌,郭慧慧,周天,等.氯离子通道5在模拟失重下磷代谢异常致骨丧失中作用的初步研究.骨科,2017,8(1): 52-56. |
氯离子通道5在模拟失重下磷代谢异常致骨丧失中作用的初步研究 |
Roles of chloride channel 5 in the bone loss caused by phosphorus metabolic abnormalities under simulated microgravity and preliminary mechanism |
投稿时间:2016-07-15 |
DOI:10.3969/j.issn.1674-8573.2017.01.013 |
中文关键词: 氯离子通道 牙本质基质蛋白1 模拟失重 骨细胞 |
英文关键词: Chloride channel 5 Dentin matrix protein 1 Microgravity Osteocyte |
基金项目:国家自然科学基金(31170898);回国人员启动基金(HG3101) |
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中文摘要: |
目的 探讨氯离子通道5(chloride channel 5, CLC5)在模拟失重下磷代谢异常致骨丧失的作用机制,为失重性骨代谢异常的防治提供数据参考。方法 ①利用回转器对IDG-SW3骨细胞模拟失重,将骨细胞分为模拟失重组(MG组)和失重对照组(MG-CON组),2 d后采用Real-time PCR检测牙本质基质蛋白1(dentinmatrix protein 1, DMP1)和CLC5在IDG-SW3骨细胞中的表达情况;②利用尾部悬吊法对小鼠模拟失重,将10只1月龄C57BL/6雌性小鼠随机分为悬尾组(SUS组)和空白对照组(CON组),每组5只,4周后取两组小鼠胫骨制作石蜡切片进行CLC5免疫组织化学染色;③2月龄DMP1基因敲除鼠和DMP1转基因鼠各5只(体重为20~25 g),制备胫骨石蜡切片,进行CLC5免疫组织化学染色。结果 Real-time PCR检测结果显示2D回转后DMP1和CLC5在IDG-SW3骨细胞中的表达量均升高,差异均有统计学意义(均P<0.05);免疫组织化学染色显示CLC5在CON组和SUS组胫骨细胞胞质和基质中均有表达,且SUS组表达量明显高于CON组,差异有统计学意义(P<0.05);CLC5在DMP1基因敲除鼠和DMP1转基因鼠骨细胞中的表达也较CON组升高,差异均有统计学意义(均P<0.05)。结论 模拟失重下CLC5参与了DMP1对磷代谢的调控,但其具体作用机制有待于进一步研究。 |
英文摘要: |
Objective To investigate the mechanisms of chloride channel 5 (CLC5) in the bone loss caused by phosphorus metabolic abnormalities under simulated microgravity. Methods The clinostat was an effective tool to simulate microgravity. Osteocytes in clinostats were divided into two groups: the microgravity group (MG) and control group (MG-CON). Real-time PCR was employed to test the mRNA expression of DMP1 and CLC5 after 2 days. Five 1-month-old C57BL/6 female mice were suspended by tail for 1 month. Another 5 mice served as controls. Immunohistochemical staining was used to detect the expression of CLC5 in the tibia, meanwhile, CLC5 expression was also detected in 2-month-old DMP1 overexpression mice and DMP1 knockout (KO) mice. Results Real-time PCR showed that the expression of DMP1 and CLC5 was increased in osteocytes under simulated microgravity (P<0.05). In addition, immunohistochemical study showed that the expression of CLC5 in tibia was also increased in unloaded mice, DMP1 overexpression mice and DMP1 KO mice. Conclusion The results indicate that CLC5 acts as a sensitive molecular mechanic under simulated microgravity. Moreover, CLC5 involved in the regulation of phosphorus metabolism by DMP1 and the further research of specific mechanism was needed. |
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