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氟对大鼠血、肝、肾和脑中5-甲基胞嘧啶表达的影响

Effect of fluoride on the expression of 5-methylcytosine in blood, liver, kidney and brain of rats

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目的 探讨氟暴露剂量和暴露时间对大鼠血、肝、肾和脑中5-甲基胞嘧啶(5-methylcytosine,5-mC)表达的影响,了解氟对不同组织DNA甲基化水平的影响是否存在差异.方法 雄性SPF级3周龄Wistar大鼠80只,按体重[(82.34±10.60)g]采用随机数字表法分为4组,每组20只.对照组饮用蒸馏水,染氟组分别饮用含氟剂量为25、50和100 mg/L的蒸馏水,自由饮食,分别于饲养1个月和3个月(n=10)时处死,采集外周血和组织样本.观察大鼠氟斑牙发生率;离子选择电极法检测骨氟和尿氟含量;酶联免疫吸附试验(ELISA)法测定血、肝、肾和脑中5-mC含量.采用析因设计方差分析方法,分析氟暴露剂量和暴露时间对大鼠外周血和不同组织中5-mC的单独作用和交互作用.结果 在饲养1个月和3个月时,对照组大鼠无氟斑牙检出,25、50、100 mg/L染氟组大鼠均有不同程度氟斑牙发生;氟暴露剂量和暴露时间均对大鼠骨氟含量[1个月:(324.985±127.094)、(846.148±331.861)、(1 886.601±250.140)、(2 420.971±135.883)mg/kg;3个月:(417.591±88.324)、(1 582.243±347.975)、(2 163.519±614.932)、(2 755.434±265.370)mg/kg]存在单独效应(F=96.692、13.077,P均<0.01),但二者间不存在交互效应(F=2.013,P>0.05);氟暴露剂量对大鼠尿氟含量存在单独效应(F=62.358,P< 0.01),暴露时间对其无单独效应(F=0.862,P>0.05),且二者间不存在交互效应(F=0.081,P>0.05).氟暴露剂量对大鼠外周血中5-mC含量存在单独效应(F=8.446,P<0.01),暴露时间对其无单独效应(F=0.095,P>0.05),但二者间存在交互效应(F=4.676,P<0.01);氟暴露剂量和暴露时间对大鼠肝脏中5-mC含量均存在单独效应(F=4.737、7.064,P<0.01或<0.05),且二者间存在交互效应(F=8.302,P< 0.01);氟暴露时间对大鼠肾中5-mC含量存在单独效应(F=6.340,P<0.05),暴露剂量对其无单独效应(F=0.140,P> 0.05),且二者间不存在交互效应(F=1.269,P>0.05);氟暴露剂量和暴露时间对大鼠脑中5-mC含量均无单独效应(F=0.633、2.065,P均>0.05).结论 氟暴露剂量和暴露时间对大鼠血、肝、肾和脑中5-mC表达水平的影响有差异,提示氟对不同组织DNA甲基化水平的影响存在差异性.
Objective To investigate the effects of fluoride exposure dose and exposure time on the expression of 5-methylcytosine (5-mC) in blood,liver,kidney and brain of rats;and to understand whether there is a difference in the effects of fluoride on DNA methylation levels in different tissues.Methods Eighty three-week old SPF male Wistar rats were randomly divided into four groups according to body weight [(82.34 ± 10.60) g],with 20 rats in each group.The rats of control group drank distilled water and the fluoride group's drank distilled water containing 25,50 and 100 mg/L of F ion,respectively.Rats were sacrificed after fed for 1 month and 3 months (n =10),and peripheral blood and tissue samples were collected.The incidence of dental fluorosis was observed in rats.Bone and urine fluoride content was detected by ion selective electrode method.The content of 5-mC in blood,liver,kidney and brain was detected by enzyme-linked immunosorbent assay (ELISA).The independent and interactive effects of fluoride exposure dose and exposure time on 5-mC in rat peripheral blood and different tissues were analyzed by factorial design anova.Results After feeding for 1 month and 3 months,all rats in the fluoride group had dental fluorosis with different severities,while none dental fluorosis was found in the control groups.Fluoride exposure dose and exposure time had a main effect on bone fluoride contents [1 month:(324.985 + 127.094),(846.148 ± 331.861),(1 886.601 + 250.140),(2 420.971 + 135.883) mg/kg;3 months:(417.591 ± 88.324),(1 582.243 ± 347.975),(2 163.519 ± 614.932),(2 755.434 ± 265.370)mg/kg;F =96.692,13.077,P < 0.01],respectively,but there was no interaction effect (F =2.013,P > 0.05);fluoride exposure dose had a main effect on urinary fluoride contents (F =62.358,P < 0.01),the exposure time had no effect on it (F =0.862,P > 0.05),and there was no interaction effect (F =0.081,P > 0.05).Fluoride exposure dose had a main effect on the 5-mC content in the blood (F =8.446,P < 0.01),the exposure time had no effect on it (F =0.095,P >0.05),and there had an interaction effect (F =4.676,P < 0.01).Fluoride exposure dose and exposure time had a main effect on the 5-mC content in the liver,respectively (F =4.737,7.064,P < 0.01 or < 0.05),and an interaction effect was exist (F =8.302,P < 0.01).Fluoride exposure time had a main effect on the 5-mC content in the kidney (F =6.340,P < 0.05),the exposure dose had no effect on it (F =0.140,P > 0.05),and there was no interaction effect (F =1.269,P > 0.05).Fluoride exposure dose and exposure time had no effect on 5-mC content in the brain (F =0.633,2.065,P > 0.05).Conclusion Fluoride exposure dose and exposure time have the different effect on the levels of 5-mC in blood,liver,kidney and brain,suggesting that there may be differences in the effects of fluoride on DNA methylation levels in different tissues.

Fluorine5-methylcytosineDNA methylation

姚英杰、马永政、仲南、裴俊瑞

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哈尔滨医科大学中国疾病预防控制中心地方病控制中心大骨节病防治研究所国家卫健委病因流行病学重点实验室黑龙江省普通高校病因流行病学重点实验室 150081

5-甲基胞嘧啶 DNA甲基化

国家自然科学基金国家自然科学基金哈尔滨医科大学伍连德青年基金黑龙江省博士后启动基金

8177346881302389WLD-QN1703LBH-Q17092

2019

中华地方病学杂志
中华医学会,哈尔滨医科大学

中华地方病学杂志

CSTPCDCSCD北大核心
影响因子:1.503
ISSN:2095-4255
年,卷(期):2019.38(9)
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