Effects of embryonic exposure to fluoride on bone

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Effects of embryonic exposure to fluoride on bone

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Note: Zebrafish are established models for endocrine disruption.

胚胎期氟暴露对斑马鱼骨骼发育的影响

Effects of embryonic exposure to fluoride on bone development of zebrafish

李俊骏 ; 哈尔滨医科大学中国疾病预防控制中心地方病控制中心地氟病防治研究所国家卫生计生委病因流行病学重点实验室(23618504)黑龙江省普通高校病因流行病学重点实验室 ; 李俊骏 ; 赵巧湜 ; 李悦 ; 包莹 ; 李冰洋 ; 严画竹 ; 霍思梦 ; 范玉梅 ; 杨艳梅 ; 高彦辉 ; 哈尔滨医科大学中国疾病预防控制中心地方病控制中心地氟病防治研究所国家卫生计生委病因流行病学重点实验室(23618504)黑龙江省普通高校病因流行病学重点实验室,哈尔滨,150081 ; 哈尔滨医科大学中国疾病预防控制中心地方病控制中心中心实验室,哈尔滨,150081

中华地方病学杂志 ; 2018年 01期 (2018 / 03 / 02) , P24 - 29

目的 研究胚胎期氟暴露对斑马鱼骨骼发育的影响.方法 将受精3 d(3 days post fertilization,3 dpf)的斑马鱼胚胎暴露于常规养鱼水(对照组,氟离子含量<0.2 mg/L)和含25、50、100 mg/L氟化钠(NaF)的养鱼水中,28℃培养5 d(8 dpf)至斑马鱼头部骨骼形成,用氟离子选择电极法检测斑马鱼胚胎整体氟含量.重新制作氟暴露模型,分别为对照(0.0 mg/L)组,低剂量暴露(0.5、1.0、4.0 mg/L)组和高剂量暴露(50.0、100.0mg/L)组,计算各组斑马鱼胚胎存活率;在40倍胚胎镜下观察斑马鱼胚胎大体形态;采用茜素红染色法对斑马鱼骨骼进行染色,显微镜下观察、数码成像后定量检测骨骼染色区域面积及累积光密度(IOD),用于分析骨质硬化和骨质疏松情况.结果对照组,25、50、100 mg/L染氟组斑马鱼胚胎整体氟含量分别为(0.32±0.01)、(0.63±0.01)、(0.86±0.02)、(1.21±0.01)μg/150条.与对照组比较,各染氟组斑马鱼胚胎整体氟含量明显增加(P均< 0.05),且有一定的剂量反应关系.重新制作氟暴露模型后,对照组和各染氟组的斑马鱼胚胎存活率分别为96.67%、96.67%、96.67%、98.33%、98.33%和98.33%,组间比较差异无统计学意义(x2=7.309,P>0.05);胚胎镜下各染氟组斑马鱼胚胎脊柱均未出现明显畸形;斑马鱼颅骨茜素红染色面积对照组为84 380.51±11 711.41,低剂量暴露(0.5、1.0、4.0 mg/L)组为92 592.16±7 143.81、92 164.85±10 136.18和95 112.26±13 721.91,均高于对照组(P均<0.05),高剂量暴露(50.0、100.0 mg/L)组为67 778.92±8 597.11和64 272.93±9 302.57,均低于对照组(P均<0.05);斑马鱼颅骨茜素红染色IOD对照组为25 094.13±6 571.86,低剂量暴露(0.5、1.0、4.0 mg/L)组为29 754.95±3 836.45、28 747.36±4 677.86和30 776.49±5 589.63,高于对照组(P均< 0.05),高剂量暴露(50.0、100.0 mg/L)组为19 263.10±4 754.72和18 202.58±4 897.15,低于对照组(P均< 0.05).结论低剂量氟暴露使斑马鱼胚胎骨质硬化,高剂量氟暴露使斑马鱼胚胎骨质疏松.

"Effects of embryonic exposure to fluoride on bone development of zebrafish"

Objective:

To study the effect of fluoride exposure on bone growth in zebrafish.

Methods:

The zebrafish larvaes at 3 days post fertilization (3 dpf) were exposed to the conventional fish water and 25,50,100 mg/L of NaF for 5 days until the skeletal bone was formed (8 dpf) and the temperature was kept at 28 ℃.The fluoride content of zebrafish embryos was detected by F-ion selective electrode.The fluoride exposure model was re-established as the control group (0.0 mg/L),the low doses group (0.5,1.0,4.0 mg/L) and the high doses group (50.0,100.0 mg/L).The survival rates of the zebrafish embryos were calculated and the morphology of zebrafish embryos was observed under 40 times microscope.The zebrafish skeleton was stained with alizarin red.The staining areas and the integrated optical density (IOD) of the bone staining were quantitatively analyzed by digital microscope to analyze the sclerotic and osteoporosis of the skull.

Results: The fluoride contents of the control group and 25,50,100 mg/L NaF groups were (0.32 ± 0.01),(0.63 ± 0.01),(0.86 ± 0.02) and (1.21 ± 0.01) μg/150 embryos. Compared with the control group,the fluoride contents of zebrafish embryos in fluoride exposed groups were increased (P < 0.05),and the dose-response relationship was obvious.The survival rates of zebrafish embryos in control group and fluoride exposed groups were 96.67%,96.67%,96.67%,98.33%,98.33% and 98.33%.There was no significant difference among different groups (x2 =7.309,P > 0.05);under a 40 times microscope,there were no obvious deformities of the spin in different groups;the areas of the alizarin red staining of the skull were 84 380.51 ± 11 711.41 in the control group,92 592.16 ± 7 143.81,92 164.85 ± 10 136.18 and 95 112.26 ± 13 721.91 in the low doses exposure groups (0.5,1.0,4.0 mg/L NaF),67 778.92 ± 8 597.11 and 64 272.93 ± 9 302.57 in the high doses groups (50.0,100.0 mg/L NaF).The areas of the alizarin red staining of the skull in the low doses exposure groups were significantly higher than that of the control group (P < 0.05),while the high doses exposure groups were lower (P < 0.05);the IOD of the alizarin red staining of the skull was 25 094.13 ± 6 571.86 in the control group,29 754.95 ± 3 836.45,28 747.36 ± 4 677.86 and 30 776.49 ± 5 589.63 in the low doses exposure groups (0.5,1.0,4.0 mg/L NaF),19 263.10 ± 4 754.72 and 18 202.58 ± 4 897.15 in the high doses groups (50.0,100.0 mg/L NaF).The IOD of the alizarin red staining of the skull in the low doses exposure groups was significandy higher than that of the control group (P < 0.05),while the high doses exposure groups was lower (P < 0.05).

Conclusion: Low doses of fluoride exposure may cause bone sclerosis in zebrafish embryos,while the high dose of fluoride exposure may cause osteoporosis.
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