Effect of different iodide intake during pregnancy and lactation on thyroid and cardiovascular function in maternal and offspring rats.

Xiaomei Yao, Xiuxiu Zhao, Hai Zhao, Yue Sun, Yue Zhang
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Abstract

OBJECTIVE We aimed to investigate the impact of different iodide intake during pregnancy and lactation on iodine concentration in urine and serum, fatty acid metabolism, thyroid and cardiovascular function in maternal and offspring rats. METHODS Pregnant rats were randomly assigned to four groups: normal adult iodide intake (NAI, 7.5 μg/d), normal pregnant iodide intake (NPI, 12.5 μg/d), 5 times (5 HI, 62.5 μg/d) and 10 times higher-than-normal pregnant iodide intake (10 HI, 125 μg/d). The maternal rats were continuously administered potassium iodide until postnatal day 16 (PN16). Thyroid function was measured by enzyme-linked immunosorbent assay (ELISA). The iodine concentration in urine and serum were detected by inductively coupled plasma mass spectrometry (ICP-MS). The messenger ribonucleic acid (mRNA) expressions of Krüppel-like factor 9 (KLF9) and thioredoxin reductase 2 (Txnrd2) were measured using quantitative real-time polymerase chain reaction (RT-qPCR). Characteristic distribution of KLF9 expression and its interaction with TRβ was assessed by immunohistochemical and immunofluorescence staining. Serum fatty acids were analyzed by Liquid Chromatography-Mass Spectrometry (LC-MS). Cardiac function and blood pressure were measured by echocardiography and a non-invasive tail-cuff system. RESULTS High iodide intake (5 HI and 10 HI) during pregnancy and lactation results in increased urinary iodine concentration (UIC), serum total iodine concentration (STIC) and serum non-protein-bound iodine concentration (SNBIC) in both maternal and offspring rats, along with significantly increased FT3 and its target gene expression of KLF9. In maternal rats of both 5 HI and 10 HI groups, systolic blood pressure (SBP) was significantly higher, the increased SBP was significantly correlated with the increased UIC (r = 0.968, p = 0.002; r = 0.844, p = 0.035), KLF9 (r = 0.935, p = 0.006; r = 0.954, p = 0.003) and the decreased Txnrd2 (r = -0.909, p = 0.012; r = -0.912, p = 0.011). In maternal rats of 10 HI group, cardiac hyperfunction with increased LVEF, LVFS and decreased LVESD were observed. The increased LVEF and decreased LVESD were significantly correlated with UIC, STIC and SNBIC (r = 0.976, p = 0.001; r = 0.945, p = 0.005; r = 0.953, p = 0.003; r = -0.917, p = 0.01; r = -0.859, p = 0.028; r = -0.847, p = 0.033), LVEF, LVFS and LVESD were significant correlated with KLF9 (r = 0.950, p = 0.004; r = 0.963, p = 0.002; r = -0.990, p = 0.0002) and Txnrd2 expression (r = -0.979, p = 0.001; r = -0.915, p = 0.01; r = 0.933, p = 0.007), and the decreased LVESD was correlated with decreased epoxyeicosatrienoic acid (EET) metabolites: 5,6-EET, 8,9-DHET and 11,12-DHET (r = 0.999, p = 0.034; r = 1.000, p = 0.017; r = 1.000, p = 0.017). While in offspring rats, no significant change in SBP and cardiac function was found. STIC and SNBIC were much lower than those in maternal rats, and eicosapentaenoic acid (EPA) metabolites (9-HEPE, 15-HEPE and 14,15 DiHETE) were significantly increased. CONCLUSION In addition to thyroid hormones, STIC, SNBIC, KLF9, Txnrd2, EET and EPA metabolites might be promising biomarkers in high iodide intake-induced thyroid and cardiovascular function.
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妊娠和哺乳期不同碘摄入量对母鼠和子鼠甲状腺和心血管功能的影响。
目的探讨孕期和哺乳期不同碘摄入量对母鼠和子鼠尿液和血清碘浓度、脂肪酸代谢、甲状腺和心血管功能的影响。方法将妊娠大鼠随机分为正常成年碘摄入量(NAI, 7.5 μg/d)、正常妊娠碘摄入量(NPI, 12.5 μg/d)、高于正常妊娠碘摄入量5倍(5 HI, 62.5 μg/d)和10倍(10 HI, 125 μg/d) 4组。母鼠连续给予碘化钾,直至产后16天(PN16)。采用酶联免疫吸附试验(ELISA)检测甲状腺功能。采用电感耦合等离子体质谱法(ICP-MS)检测尿、血清碘浓度。采用实时定量聚合酶链式反应(RT-qPCR)检测kr pel样因子9 (KLF9)和硫氧还蛋白还原酶2 (Txnrd2)信使核糖核酸(mRNA)的表达。采用免疫组织化学和免疫荧光染色评价KLF9表达的特征分布及其与TRβ的相互作用。采用液相色谱-质谱法(LC-MS)分析血清脂肪酸。通过超声心动图和无创尾袖系统测量心功能和血压。结果妊娠和哺乳期高碘摄入(5 HI和10 HI)导致母鼠和子鼠尿碘浓度(UIC)、血清总碘浓度(STIC)和血清非蛋白结合碘浓度(SNBIC)升高,FT3及其靶基因KLF9表达显著升高。5 HI组和10 HI组母鼠收缩压(SBP)均显著升高,SBP升高与UIC升高有显著相关(r = 0.968, p = 0.002;r = 0.844, p = 0.035), KLF9 (r = 0.935, p = 0.006;r = 0.954, p = 0.003),减少Txnrd2 (r = -0.909, p = 0.012;R = -0.912, p = 0.011)。10 HI组母鼠心肌功能亢进,LVEF升高,LVFS升高,LVESD降低。LVEF升高、LVESD降低与UIC、STIC、SNBIC显著相关(r = 0.976, p = 0.001;R = 0.945, p = 0.005;R = 0.953, p = 0.003;R = -0.917, p = 0.01;R = -0.859, p = 0.028;r = -0.847, p = 0.033), LVEF、LVFS、LVESD与KLF9显著相关(r = 0.950, p = 0.004;R = 0.963, p = 0.002;r = -0.990, p = 0.0002)和Txnrd2表达式(r = -0.979, p = 0.001;R = -0.915, p = 0.01;r = 0.933, p = 0.007), LVESD降低与环氧二碳三烯酸(EET)代谢物5,6-EET、8,9- dhet和11,12- dhet减少相关(r = 0.999, p = 0.034;R = 1.000, p = 0.017;R = 1.000, p = 0.017)。而子代大鼠收缩压和心功能未见明显变化。STIC和SNBIC明显低于母鼠,二十碳五烯酸(EPA)代谢产物(9-HEPE、15- hepe和14,15 DiHETE)显著升高。结论除甲状腺激素外,STIC、SNBIC、KLF9、Txnrd2、EET和EPA代谢物可能是高碘摄入诱导甲状腺和心血管功能的有前景的生物标志物。
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