亚慢性氟离子饮水染毒对小鼠肾脏毒性作用及肾组织STIM1和ORAI1表达的影响  

Effects of subchronic fluoride exposure through drinking water on renal toxicity and the expressions of STIM1 and ORAI1 in renal tissues in mice

作　　者：吴璇 丁凡 刘盈莹 吴庆[1] WU Xuan;DING Fan;LIU Yingying;WU Qing(School of Public Health/Key Laboratory of Public Health Safety of Ministry of Education,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学,公共卫生学院/公共卫生安全教育部重点实验室,上海200032

出　　处：《环境与职业医学》2022年第5期493-498,共6页Journal of Environmental and Occupational Medicine

基　　金：中国科学院中国-斯里兰卡联合科教中心资助项目(无编号)。

摘　　要：[背景]氟可以通过诱导胞内钙超载导致细胞损伤。钙池操纵性钙内流(SOCE)对于维持胞内钙稳态具有重要作用,但氟对肾组织SOCE的影响是未知的。[目的]探究氟离子经饮水亚慢性染毒对小鼠的肾脏毒性,以及对肾脏SOCE关键蛋白中的基质相互作用分子1(STIM1)和钙释放激活钙通道调节分子1(ORAI1)表达的影响。[方法]将20只雄性ICR小鼠随机分为4组,每组5只:分别为0(对照组)、0.3、3和30 mg·L的氟离子染毒组,饮水染毒12周。测定染毒后小鼠的体重及肝肾脏器系数;观察小鼠肾脏的组织病理学改变;收集小鼠染毒12周末的24 h尿液,测定尿肌酐(UCr)、尿钙(UCa)、白蛋白(ALB)和β2-微球蛋白(β2-MG)的水平;使用蛋白质免疫印迹法检测肾脏中STIM1和ORAI1的蛋白表达水平;通过STIM1和ORAI1荧光共定位进一步验证STIM1和ORAI1的表达水平。[结果]氟离子染毒后,各组间小鼠的体重及肝肾脏器系数差异无统计学意义(均P> 0.05)。光镜下,3、30 mg·L的氟离子组中,小鼠肾脏组织可见肾小管细胞变性、顶端突出、脱落和扩张等;小鼠尿液指标可见,各组间小鼠UCr差异无统计学意义(P> 0.05);与对照组相比,3、30 mg·L氟离子组中UCr校正后的UCa水平分别为(0.075±0.014)、(0.081±0.012)mol·mo(以每摩尔UCr表示),虽略有升高趋势,但差异无统计学意义;ALB水平在各组间差异也无统计学意义(P> 0.05);不同染毒组β2-MG水平有差异,30 mg·L氟离子组的β2-MG水平为(0.077±0.014)g·mol,较对照组升高(P <0.05);蛋白质免疫印迹法结果显示,各组间STIM1和ORAI1水平差异均具有统计学意义(F=18.411、6.853,P=0.001、0.013);与对照组相比,3、30 mg·L氟离子组中STIM1蛋白表达水平升高(P <0.05),30 mg·L氟离子组ORAI1蛋白表达水平升高(P <0.05);STIM1和ORAI1荧光共定位可见,3、30 mg·L氟离子组STIM1和ORAI1的表达上调。[结论]氟离子经饮水亚慢性染毒可以上调肾组织STIM1和ORAI1表达[Background] It has been found that fluoride may cause cell damage by inducing intracellular calcium overload. Store-operated calcium entry(SOCE) plays an important role in maintaining intracellular calcium homeostasis, but the effect of fluoride on renal SOCE is unknown.[Objective] To explore the renal toxicity and the expression levels of the key proteins of SOCE,stromal interaction molecule 1(STIM1) and calcium release-activated calcium modulator 1(ORAI1) in the kidney tissues of mice exposed to fluoride subchronically.[Methods] Twenty male ICR mice were randomly divided into four groups with five mice in each group, including 0(control group), 0.3, 3, and 30 mg·Lfluoride groups. The mice were given drinking water containing designed fluoride for 12 weeks. Body weight and liver and kidney organ coefficients of the mice were measured after the exposure;histopathological changes of the mouse kidney were observed;24 h urine was collected at the end of 12 weeks of exposure to determine the levels of urine creatinine(UCr), urine calcium(UCa), albumin(ALB), and β2-microglobulin(β2-MG);the protein expression levels of STIM1 and ORAI1 in the kidney were detected by Western blotting;the fluorescence co-localization of STIM1 and ORAI1 was used to further verify the expression levels of STIM1 and ORAI1.[Results] After the exposure, there were no differences in body weight as well as liver and kidney organ coefficients among the groups(P > 0.05). Under optical microscope, the renal tubular cell showed degeneration, apical protrusion, shedding, and dilation in the 3 and30 mg·Lfluoride groups. There was no statistical difference in UCr among the mice in each group(P > 0.05). Compared with the control group, the levels of UCa adjusted by UCr in the 3 and 30 mg·Lfluoride groups were(0.075±0.014) and(0.081±0.012) mol·mol-1(represent by UCr per mol), which had a rising trend but showed no statistical difference. No difference was identified in the level of ALB among the groups(P > 0.05). The levels of β2-MG showed diff

关 键 词：氟 亚慢性染毒 肾毒性 钙池操纵性钙内流 基质相互作用分子1 钙释放激活钙通道调节分子1 

分 类 号：R114[医药卫生—卫生毒理学]