摘要
目的了解妊娠期至成年期氟铝联合暴露对子代大鼠骨组织中miR-21及血清中抗酒石酸酸性磷酸酶-5b(TRACP-5b)的影响。方法将36只健康成年清洁级SD孕鼠随机分为9组,分别为对照(自来水)组和氟化钠(60、240mg/L)、氯化铝(600、1 000 mg/L)染毒组及氟化钠+氯化铝联合染毒组,每组4只。采用自由饮水方式染毒,母鼠染毒从妊娠第0天至子代大鼠出生第21天(断乳);断乳后从每组随机选取8只子代大鼠(雌雄各半)延续同组剂量染毒至出生后第90天。采用氟离子选择电极法测定尿氟、骨氟,采用石墨炉原子吸收光谱法测定尿、骨中铝含量,采用实时荧光定量PCR (qRT-PCR)检测骨组织中miR-21的表达水平。采用固相夹心ELISA法检测血清中抗酒石酸酸性磷酸酶-5b(TRACP-5b)的含量。结果与对照组比较,除单独氯化铝染毒组和60 mg/L NaF+1 000 mg/L氯化铝染毒组外,其余染毒组仔鼠的尿氟浓度均升高(P<0.05);除单独NaF染毒组和240 mg/L NaF+600 mg/L氯化铝染毒组外,其余染毒组仔鼠的尿铝浓度均升高(P<0.05);除单独氯化铝染毒组和60 mg/L NaF+1 000 mg/L氯化铝染毒组外,其余染毒组仔鼠骨氟含量均升高;单独氯化铝染毒组和240 mg/L NaF+1 000 mg/L氯化铝染毒组仔鼠的骨铝含量均升高(P<0.05)。尿氟浓度在各浓度NaF+氯化铝联合染毒组中均表现为拮抗作用(P<0.05);尿铝浓度在60 mg/L NaF+1 000 mg/L氯化铝联合染毒组中均表现为协同作用,在其余各浓度NaF+氯化铝联合染毒组中均表现为拮抗作用(P<0.05);骨氟含量在各浓度NaF+氯化铝联合染毒组中均表现为拮抗作用(P<0.05);骨铝含量在各浓度NaF+氯化铝联合染毒组中未见交互作用(P>0.05)。与对照组比较,240 mg/L NaF染毒组和60 mg/L NaF+1 000 mg/L氯化铝染毒组、240 mg/L NaF+1 000 mg/L氯化铝染毒组仔鼠血清中TRACP-5b的水平均降低(P<0.05);60 mg/L NaF染毒组、1 000 mg/L氯化铝染毒组和60 mg/L NaF+600 mg/L氯化铝染毒组、240 mg/L NaF+600 mg/L氯化铝染毒组仔鼠骨组织中miR-21的表达水平均升高(P<0.05)。氟铝联合染毒对血清中TRACP-5b水平的影响表现为拮抗作用。氟铝联合染毒对不同性别仔鼠骨组织中miR-21表达的影响均存在交互作用(P<0.05);对不同性别仔鼠骨组织中miR-21表达的影响在60 mg/L NaF+600 mg/L氯化铝染毒组、240mg/L NaF+600 mg/L氯化铝染毒组中均表现为协同作用(P<0.05),在其余联合染毒组中表现为拮抗作用(P<0.05)。结论氟铝联合暴露可能通过升高骨组织中miR-21表达和降低血清中TRACP-5b水平,从而促进妊娠期至子代大鼠成年前仔鼠骨组织中破骨细胞的活性。
Objective To investigate the effects of exposure to fluoride combined with aluminum from pregnancy to adulthood on miR-21 in bone tissue and tartrate resistant acid phosphatase-5 b(TRACP-5 b) in serum of the offspring rats. Methods Thirtysix pregnant SD rats were randomly divided into control and experimental groups as follows,control group(drinking tap water),fluoride alone exposure group(drinking the water containing sodium fluoride of 60 mg/L and 240 mg/L respectively),aluminum alone exposure group(drinking the water containing aluminum chloride of 600 mg/L and 1 000 mg/L),fluoride+aluminum joint exposure groups. Four pregnant rats in each group. The pregnant rats were treated with fluoride and aluminum through drinking water from pregnant day 0 to postnatal day 21(PND21) of the offspring rats. Eight offspring rats selected randomly in each group were treated as above. Fluoride and aluminum in urine and bone were determined by fluorine ion electrode selection method and graphite furnace atomic absorption. Real time fluorescence quantitative PCR(qRT-PCR) was used to detect the expression of miR-21 in bone tissue. TRACP-5 b content in serum was determined by tartrate resistant acid phosphatase solid-phase sandwich ELISA method. Results Compared with the control group,except the AlCl3 alone exposure group and 60 mg/L NaF+1 000 mg/L AlCl3 joint exposure group,the fluoride concentration in urine were higher(P<0.05); Except the NaF alone exposure group and 240 mg/L NaF +600 mg/L AlCl3 joint exposure group,the aluminum concentrations in urine were higher(P <0.05);Except the AlCl3 alone exposure group and 60 mg/L NaF+1 000 mg/L Al Cl3 joint exposure group,the fluoride concentrations in bone were higher(P<0.05); The aluminium concentrations in bone in the AlCl3 alone groups and 240 mg/L NaF+1 000 mg/L AlCl3 joint exposure group were higher(P <0.05). The combined effects of fluoride and aluminium were antagonism on the content of fluoride in urine and bone among each joint exposure group(P <0.05), and on the content of aluminium in urine among each joint exposure group except 60 mg/L NaF+1 000 mg/L Al Cl3 joint exposure group with synergistic interaction(P<0.05),and showed no interaction on the content of aluminium in bone among each joint exposure group(P>0.05). Compared with the control group,the serum TRACP-5 b concentrations in 240 mg/L NaF alone exposure group,60 mg/L NaF+1 000 mg/L AlCl3 joint exposure group and 240 mg/L NaF+1 000 mg/L AlCl3 joint exposure group were decreased(P<0.05), the expression levels of miR-21 in the bone in 60 mg/L NaF alone exposure group,1 000 mg/L AlCl3 alone exposure group,60 mg/L NaF+600 mg/L AlCl3 joint exposure group and 240 mg/L NaF +600 mg/L AlCl3 joint exposure group were increased(P <0.05).The combined effects of fluoride and aluminium were antagonism on the TRACP-5 b concentrations in serum among each joint exposure group(P<0.05), and antagonism on the expression levels of miR-21 among each joint exposure group except 60 mg/L NaF+600 mg/L AlCl3 joint exposure group and 240 mg/L NaF +600 mg/L Al Cl3 joint exposure group identified as synergism(P <0.05).Conclusion Fluoride and aluminium joint exposure during the period from pregnancy to adulthood can reduce the TRACP-5 b level in serum and increase the levels of miR-21 in bone,it maybe promote osteoclast activity in bone of offspring rats.
作者
韦山禄
葛启迪
曾智东
张华
谢春
WEI Shan-lu;GE Qi-di;ZENG Zhi-dong;ZHANG Hua;XIE Chun(Department of Occupational and Environmental Health y,School of Public Health,Guizhou Medical University ,Guiyangt Guizhou 550025'China)
出处
《环境与健康杂志》
CAS
北大核心
2018年第7期569-574,共6页
Journal of Environment and Health
基金
国家自然科学基金(81560519)
