氟、砷染毒对大鼠破骨细胞骨吸收能力的影响及其机制

杨德山; 洪峰

doi:10.13213/j.cnki.jeom.2018.17694

氟、砷染毒对大鼠破骨细胞骨吸收能力的影响及其机制

杨德山,
洪峰

Effects of fluoride and arsenic exposure on bone absorptive capacity of osteoclasts in rats and related mechanism

摘要

摘要:
目的研究氟砷联合染毒对大鼠破骨细胞骨吸收能力的影响及其机制。

方法 54只清洁级SD大鼠随机分为9组，每组6只，雌雄各半。根据预实验及文献确定染毒剂量，用NaF和NaAsO₂分别对实验动物予以灌胃染毒，各染毒组毒物剂量及缩写分别为：对照组（F₀As₀）、低氟组（5 mg/kg，F₅）、高氟组（20 mg/kg，F₂₀）、低砷组（2.5 mg/kg，As_2.5）、高砷组（10 mg/kg，As₁₀）、低氟低砷组（5 mg/kg+2.5 mg/kg，F₅As_2.5）、低氟高砷组（5 mg/kg+10 mg/kg，F₅As₁₀）、高氟低砷组（20 mg/kg+2.5 mg/kg，F₂₀As_2.5）、高氟高砷组（20 mg/kg+10 mg/kg，F₂₀As₁₀）。酶联免疫吸附法（ELISA法）测定大鼠血清中抗酒石酸酸性磷酸酶5b（TRACP-5b）、Ⅰ型胶原吡啶交联终肽（ICTP）含量及大鼠右侧股骨组织匀浆上清液中骨保护素（OPG）、前列腺素E2（PGE₂）、1，25二羟基维生素D31，25（OH）₂D₃含量。

结果大鼠血清中ICTP、TRACP-5b蛋白表达水平在高氟组均较对照组升高（F₀As₀：1.352±0.025，2.942±0.078；F₂₀：1.427±0.026，3.202±0.142；均P < 0.05），而单独染砷组与对照组的差异均无统计学意义（Ps>0.05）。对于氟砷联合染毒组，染氟剂量在20mg/kg时，ICTP、TRACP-5b蛋白含量在高氟高砷组较高氟低砷组明显降低（F₂₀As_2.5：1.442±0.034，3.280±0.095；F₂₀As₁₀：1.345±0.042，3.053±0.084；均P < 0.05）。氟砷的偏相关分析及交互作用结果显示TRACP-5b、ICTP蛋白含量的改变与氟浓度呈正相关（偏相关系数：0.663，0.392；均P < 0.05），氟起到主效应作用（F=24.560，8.188；均P < 0.05）。大鼠骨组织OPG、PGE₂、1，25（OH）₂D₃表达水平在高氟组均较对照组升高（F₀As₀：12.278±0.220，1.352±0.025，9.582±0.363；F₂₀：14.087±0.797，1.427±0.026，11.915±0.931；均P < 0.05）。对于氟砷联合染毒组，染氟剂量为2.5 mg/kg时，低氟高砷组的PGE₂较低氟低砷组低（F₅As_2.5：1.442±0.034；F₅As₁₀：1.345±0.080；P < 0.05）；染氟剂量为20 mg/kg时，高氟高砷组1，25（OH）₂D₃较高氟低砷组低（F₂₀As_2.5：12.082±0.860；F₂₀As₁₀：10.567±0.695；均P < 0.05）。氟砷的偏相关分析及交互作用结果显示OPG、PGE₂、1，25（OH）₂D₃含量改变与氟呈正相关（偏相关系数：0.804，0.487，0.748；均P < 0.05），氟起到主效应作用（F=47.100，9.031，3.114；均P < 0.05）；砷对OPG、PGE₂、1，25（OH）₂D₃的直接影响不明显（P>0.05）。

结论砷能抑制氟暴露对大鼠破骨细胞骨吸收功能的促进作用，氟砷之间的交互作用主要表现为拮抗作用。PGE₂、1，25（OH）₂D₃可能是氟砷对破骨细胞骨吸收功能进行调控的重要因子。

Abstract:
Objective To explore the toxicity and mechanism of combined exposure to fluoride and arsenic on bone absorptive capacity of osteoclasts in rats.

Methods A total of 54 SD rats of clean grade were randomly divided into 9 groups, with 6 rats in each group (half males and half females). Based on a pilot experiment and literature review, the animals were treated with NaF and NaAsO₂ by intragastric administration:control group (F₀As₀), low fluoride group (5mg/kg, F₅), high fluoride group (20mg/kg, F₂₀), low arsenic group (2.5mg/kg, As_2.5), high arsenic group (10 mg/kg, As₁₀), low fluoride and low arsenic group (5 mg/kg+2.5 mg/kg, F₅As_2.5), low fluoride and high arsenic group (5 mg/kg+10 mg/kg, F₅As₁₀), high fluoride and low arsenic group (20 mg/kg+2.5 mg/kg, F₂₀As_2.5), and high fluoride and high arsenic group (20 mg/kg+10 mg/kg, F₂₀As₁₀). The levels of tartrateresistant acid phosphatase 5b (TRACP-5b) and pyridinoline cross linked carboxyterminal telopeptide of type Ⅰ collagen (ICTP) in rat serum, as well as the levels of osteoprotegerin (OPG), prostaglandin (PGE₂), and 1, 25-dihydroxy vitamin D31, 25(OH)₂D₃ in homogenate supernatant of right femur tissue samples were determined by ELISA.

Results The expression levels of ICTP and TRACP-5b in the high fluoride group were higher than those in the control group (F₀As₀:1.352±0.025, 2.942±0.078; F₂₀:1.427±0.026, 3.202±0.142; Ps < 0.05), while there was no significant difference between any individual arsenic exposure group and the control group (Ps>0.05). The expression levels of ICTP and TRACP-5b in the high fluoride and high arsenic group were lower than those in the high fluoride and low arsenic group (F₂₀As_2.5:1.442±0.034, 3.280±0.095; F₂₀As₁₀:1.345±0.042, 3.053±0.084; Ps < 0.05). The results of partial correlation analysis and interaction analysis showed that the expression levels of TRACP-5b and ICTP were positively correlated with fluoride concentration (partial correlation coefficient:0.663, 0.392; Ps < 0.05), and fluoride showed the main effect (F=24.560, 8.188; Ps < 0.05). The expression levels of OPG, PGE₂, and 1, 25(OH)₂D₃ in the groups treated with high doses of fluoride were significantly higher than those in the control group (F₀As₀:12.278±0.220, 1.352±0.025, 9.582±0.363; F₂₀:14.087±0.797, 1.427±0.026, 11.915±0.931; Ps < 0.05). The expression level of PGE₂ in the low fluoride and high arsenic group was lower than that in the low fluoride and low arsenic group (F₅As_2.5:1.442±0.034; F₅As₁₀:1.345±0.080; P < 0.05). The expression level of 1, 25(OH)₂D₃ in the high fluoride and high arsenic group was lower than that in the high fluoride and low arsenic group (F₂₀As_2.5:12.082±0.860; F₂₀As₁₀:10.567±0.695; Ps < 0.05). The results of partial correlation analysis and interaction analysis showed that the expression levels of OPG, PGE₂, and 1, 25(OH)₂D₃ were positively correlated with fluoride concentration (partial correlation coefficient:0.804, 0.487, 0.748; Ps < 0.05), and fluoride showed as the main effect (F=47.100, 9.031, 3.114; Ps < 0.05); the direct toxic effects of arsenic on OPG, PGE₂, and 1, 25(OH)₂D₃ were minor (P>0.05).

Conclusion Arsenic can inhibit the auxo-action of fluorine on the bone absorption of osteoclasts in rats, and the fluorine-arsenic interaction is mainly antagonistic. PGE₂ and 1, 25(OH)₂D₃ may be important factors in the regulation of osteoclast bone absorption function.

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