中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (17): 2675-2680.doi: 10.12307/2023.158

• 骨组织构建 bone tissue construction • 上一篇    

氟中毒兔股骨骨形态发生蛋白2表达与骨氟含量的关系

梁伟叶1，段庆红2，3   

  1. 1贵州医科大学，贵州省贵阳市  550004；2贵州医科大学附属肿瘤医院影像科，贵州省贵阳市  550000；3贵州医科大学附属医院影像科，贵州省贵阳市  550004
  • 收稿日期:2022-03-14 接受日期:2022-05-11 出版日期:2023-06-18 发布日期:2022-10-24
  • 通讯作者: 段庆红，博士，副教授，贵州医科大学附属肿瘤医院影像科，贵州省贵阳市 550000；贵州医科大学附属医院影像科，贵州省贵阳市 550004
  • 作者简介:梁伟叶，女，1994年生，河南省南阳市人，贵州医科大学在读硕士，主要从事地氟病研究。
  • 基金资助:
    国家自然科学基金-贵州省人民政府联合基金项目子课题(U1812403-6-1-7)，项目负责人：段庆红；贵州省科技计划项目(黔科合支撑[2021]一般451)，项目负责人：段庆红

Correlation between femur bone morphogenetic protein 2 expression and bone fluoride content in fluorosis rabbits

Liang Weiye1, Duan Qinghong2, 3   

  1. 1Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Department of Imaging, Affiliated Cancer Hospital of Guizhou Medical University, Guiyang 550000, Guizhou Province, China; 3Department of Imaging, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Received:2022-03-14 Accepted:2022-05-11 Online:2023-06-18 Published:2022-10-24
  • Contact: Duan Qinghong, MD, Associate professor, Department of Imaging, Affiliated Cancer Hospital of Guizhou Medical University, Guiyang 550000, Guizhou Province, China; Department of Imaging, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • About author:Liang Weiye, Master candidate, Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Supported by:
    the National Natural Science Foundation - Guizhou Provincial People’s Government Joint Fund Project, No. U1812403-6-1-7 (to DQH); Guizhou Science and Technology Planning Project, No. [2021]451 (to DQH)

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摘要：


文题释义：

骨形态发生蛋白：又称骨形成蛋白，是一组具有类似结构的高度保守的功能蛋白，属于转化生长因子β家族。骨形态发生蛋白单体由信号肽、前功能区和成熟肽3个部分组成，其结构类似一个扁平的弧形表面，通常包括一个长形的α螺旋和从α螺旋两端发出的两对反向平行β链。成熟的骨形态发生蛋白是由二硫键连接的同型或异型二聚体，均有分子活性，被释放到细胞外后，与靶细胞表面的相应受体结合而发挥作用。
Ⅰ型前胶原氨基端原肽：是人体最丰富的胶原蛋白形式，约占骨质的90%。Ⅰ型胶原主要在骨组织中合成，首先合成的是原胶原，在Ⅰ型原胶原的氨基N端和C端各有一个延长肽，分别被称为Ⅰ型原胶原N端前肽和Ⅰ型原胶原C端前肽；当Ⅰ型原胶原被成骨细胞分泌到细胞外时，N端和C端的延长肽会被蛋白酶切割，除少量沉积在骨基质中以外，大量会进入血液循环，最终被肝脏代谢清除。

背景：氟中毒是一种常见的地方性疾病，其严重程度主要取决于氟化物暴露剂量和暴露持续时间。骨形态发生蛋白2在氟骨症发病过程中有重要作用。
目的：探究兔股骨骨形态发生蛋白2表达与骨氟含量的关系，以及骨形成标志物骨钙素与Ⅰ型前胶原氨基端原肽在不同氟浓度下的变化。
方法：将16只新西兰白兔随机分为对照组、低氟组、中氟组和高氟组，每组4只，分别饮用含0，100，200，400 mg/L氟化钠的自来水，饲养期间动物自由饮水、进食，实验周期6个月。观察氟斑牙发生发展情况，采用灰化蒸馏-氟试剂比色法测定兔股骨中氟含量，蛋白免疫印迹法检测股骨中骨形态发生蛋白2蛋白表达，采用酶联免疫吸附实验检测血清骨钙素和Ⅰ型前胶原氨基端原肽水平。
结果与结论：①低氟组以轻度氟斑牙为主，表现为牙齿失去光泽，部分可见细纹；中氟组以中度氟斑牙为主，表现为牙齿表面凹凸不平、色泽不均，个别表现为重度氟斑牙，表现为牙齿缺损；高氟组以重度氟斑牙为主，表现为牙齿表面无光泽，牙齿发育畸形；②随着摄氟质量浓度的升高，兔股骨内骨氟含量增加，其中高氟组骨氟含量高于其他3组(P < 0.01，P < 0.001)；高氟组血清骨钙素水平高于对照组
(P < 0.05)，血清Ⅰ型前胶原氨基端原肽水平低于对照组、低氟组(P < 0.05)；③低氟组股骨内骨形态发生蛋白2表达高于对照组、高氟组(P < 0.05)；④Pearson相关分析显示，低氟组骨形态发生蛋白2表达与骨氟含量呈正相关(r=0.951，P < 0.05)，中、高氟组骨形态发生蛋白2表达与骨氟含量无显著相关(r=0.889、r=-0.116，P > 0.05)；⑤兔股骨骨形态发生蛋白2表达与骨氟含量的相关性与氟暴露浓度有关，过高或过低的氟浓度均会影响骨形态发生蛋白2的表达。 

https://orcid.org/0000-0002-7961-3525(梁伟叶)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 氟中毒, 骨氟, 骨形态发生蛋白2, 骨钙素, Ⅰ型前胶原氨基端原肽, 兔

Abstract: BACKGROUND: Fluorosis is a common endemic disease, and its severity depends on the dose and duration of fluoride exposure. Bone morphogenetic protein 2 plays an important role in the pathogenesis of skeletal fluorosis.
OBJECTIVE: To explore the relationship between the expression of bone morphogenetic protein 2 and bone fluoride content in rabbit femur and the changes of bone formation markers, osteocalcin and procollagen l N-terminal propeptide, under different fluoride concentrations.  
METHODS: Sixteen New Zealand white rabbits were randomly divided into control group, low dose group, middle dose group, and high dose group, with four rabbits in each group. Animals drank tap water containing 0, 100, 200, and 400 mg/L sodium fluoride, respectively. The experimental period was 6 months. During the feeding period, all animals had free access to water and food. Occurrence and development of dental fluorosis was observed regularly. The fluorine content in rabbit femur was determined by ashing distillation-fluorine reagent colorimetry. The expression of bone morphogenetic protein 2 in femur was detected by western blot. The levels of osteocalcin and procollagen l N-terminal propeptide in serum were detected by enzyme-linked immunosorbent assay.  
RESULTS AND CONCLUSION: The rabbits in fluoride exposure groups showed different degrees of fluorosis. In the low dose group, mild fluorosis was found and mainly manifested with tarnished teeth and some fine lines. The middle dose group was dominated by moderate dental fluorosis, manifested as uneven tooth surface and color. Some models with dental defects presented with severe dental fluorosis. The high dose group was dominated by severe dental fluorosis, which was characterized by dull tooth surface and deformed tooth development. The bone fluoride content in the femur of rabbits increased with the increase of fluoride intake, which was higher in the high dose group than the other three groups (P < 0.01, P < 0.001). The serum osteocalcin level in the high dose was higher than that in the control group (P < 0.05) and the serum level of procollagen l N-terminal propeptide was lower than that in the control group and the low dose group (P < 0.05). The expression of bone morphogenetic protein 2 in the femur was higher in the low dose group than the control and high dose groups (P < 0.05). Pearson correlation analysis showed that the expression of bone morphogenetic protein 2 in the low dose group was positively correlated with the bone fluoride content (r=0.951, P < 0.05), while there was no significant correlation between the expression of bone morphogenetic protein 2 and the content of bone fluoride in the middle and high dose groups (r=0.889, r=-0.116, P > 0.05). To conclude, the expression of bone morphogenetic protein 2 in rabbit femur is bidirectionally related to bone fluoride concent, which is related to fluoride exposure concentration, that is, too high or too low fluoride content will alter the expression of bone morphogenetic protein 2.

Key words: fluorosis, bone fluoride, bone morphogenetic protein 2, osteocalcin, procollagen l N-terminal propeptide, rabbit

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