论文部分内容阅读
[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,As2.5)、高砷组(10 mg/kg,As10)、低氟低砷组(5mg/kg+2.5 mg/kg,F5As2.5)、低氟高砷组(5mg/kg+10 mg/kg,F5As10)、高氟低砷组(20 mg/kg+2.5 mg/kg,F20As2.5)、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D3[1,25(OH)2D3]含量.[结果]大鼠血清中ICTP、IRACP-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(F20As2.5:1.442±0.034,3.280±0.095;F20As10:L345±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(FsAs2.5:1.442±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(F20As2.5:12.082±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、l,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.“,”[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 (FoAso),low fluoride group (5 mg/kg,F5),high fluoride group (20 mg/kg,F20),low arsenic group (2.5 mg/kg,As2.5),high arsenic group (10 mg/kg,As10),low fluoride and low arsenic group (5 mg/kg+2.5 mg/kg,F5As2s),low fluoride and high arsenic group (5 mg/kg+10mg/kg,F5As10),high fluoride and low arsenic group (20mg/kg+2.5 mg/kg,F20Asz5),and high fluoride and high arsenic group (20mg/kg+10mg/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 D3 [1,25(OH)2D3] in homogenate supematant of fight 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 (FoAs0:1.352 ± 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 (F20As2.5:1.442 ± 0.034,3.280 ± 0.095;F2oAs10:1.345 ± 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 (FoAs0:12.278 ± 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 (F5As2.5:1.442 ± 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 (F20As2.5:12.082 ± 0.860;F2oAs1o: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 anxo-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.