2002 - Effects of various doses of I and F on rats

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2002 - Effects of various doses of I and F on rats

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184. 国秀娟;氟过量与碘氟过量对甲状腺功能和形态影响的实验研究[D];中国医科大学;2002年
Guo Xiujuan - "Experimental Study on the Effects of Excessive Fluoride and Iodine-Fluoride Combination on Thyroid Function and Morphology" (2002)
(This doctoral dissertation, authored by Guo Xiujuan, explores the experimental research on the impacts of excessive fluoride and a combination of iodine-fluoride on thyroid function and morphology. The study was conducted at China Medical University in the year 2002.)
http://cdmd.cnki.com.cn/Article/CDMD-10 ... 092330.htm
http://poisonfluoride.com/phpBB3/viewto ... =21&t=1537

前言 碘与氟是人体必需的两种微量元素,机体对碘、氟的摄取主要取决于外环境。本课题组于实现全民食盐碘化(universal salt iodated USI)法规3年后对辽宁、河北三个不同碘摄入量农村社区(人群尿碘中位数分别为103μg/L、374μg/L、615μg/L)进行甲状腺疾病的流行病学调查,结果显示:碘摄入量增加导致甲状腺功能减退症、自身免疫甲状腺炎显著增加,乳头状甲状腺癌患病率增加。同时我们发现,在高碘地区存在高氟的因素,氟过量可引起机体不同程度的代谢紊乱和中毒表现,如氟斑牙、氟骨症,氟过量对甲状腺的影响是国际学者长期争论的问题,有关于此的流行病学调查和动物实验研究结果亦不一致,氟对甲状腺影响机制尚不明确。本实验拟通过对大鼠碘氟代谢,甲状腺激素水平测定及形态学的观察来探讨氟过量及碘氟过量对甲状腺的影响,以明确氟在甲状腺损伤中的作用,更好地了解微量元素之间的复杂关系和对人类健康的影响,为防治措施提供理论依据。 实验材料 一、实验动物 选用我校实验动物部提供的5周龄Wistar大鼠160只,体重125~165g,雌雄各半,随机分8组。实验期间饲以普通饲料和去离子水。 二、主要仪器 电子天平;自动体重秤;不锈钢代谢笼;熔蜡箱;水浴锅;温箱;pH计;-20℃冰箱;-70℃冰箱;石蜡切片机;离心机;透射电镜;光学显微镜;显微图像分析仪;万能显微镜。 实验方法 一、动物分组与处理 将5周龄WISthe大鼠160只,体重125J,随机分为8组, 每组20只,雌雄各半。选用动物部提供的普通饲料,分别自由饮 用不同浓度的氟化钠配制的去离子水,碘过量组为同一浓度,用碘 酸钾配制。分组如下: 对照组:去离子水 氟过量二组:含氟15ppm的去离子水 (相当于3倍高氟) 氟过量11组:含氟30ppm的去离子水 (相当于6倍高氟) 氟过量皿组:含氟60ppm的去离子水 (相当于12倍高氟) 碘过量组:含碘1200pglL的去离子水 (相当于6倍高碘) 碘氟过量互组:含碘1200pgiL、氟15ppm的去离子水 (相当于6倍高碘3倍高氟) 碘氟过量D组:含碘 1200pg/L、氟 30ppm的去离子水 (相当于6倍高碘在倍高氟) 碘氟过量皿组:含碘1200Pg/L、氟60PPm的去离子水 (相当于6倍高碘* 倍高氟) 实验早期第75天处死一半动物,每组10只,后期第150天处 死剩余大鼠。 二、标本收集及处理 二.尿 2.体重 3.血清 ·2· 4.甲状腺及其湿重 5.常规组织形态学观察 O)石蜡切片的制作过程 固定;脱水;透明;浸蜡;包埋;切片。 (2)HE染色 脱蜡;水化;苏木精染色;二%盐酸酒精分化;返蓝;伊红染色; 水洗;脱水;透明;封片。 6.透射电镜超微结构观察 三、血清垂体——甲状腺激素水平 ’IT、’IT用免疫放射法(RIAXhH采用固相免疫放射分析法 (IRMA)。 四、尿碘尿氟测定 尿碘采用砷钵催化分光光度法,尿氟采用氟离子选择电极法。 五、图像采集与分析 六、数据统计学处理 应用SPSS10.0统计软件进行单因素方差分析*值检验,进 一步比较用 SNK检验*检验L结果用 k。SD表示,以 P<0.05 为有统计学意义。 实验结果 一、尿碘测定结果 碘过量组及碘氟过量组75天上50天尿碘水平与对照组比较 明显升高帅<0.05人碘氟过量各组与碘过量组比较无显著差异 (P>0.05)。 二、尿氟测定结果’ 氟过量皿组75天,碘氟过量皿组的尿氟水平与对照组比较明 显升高帅<0.05入其 150天水平与对照组比较明显增加h<0· 05入与 75天水平比较亦明显升高h<0刀5入碘氟过量 I、匝组 ·3· 与对照组比较无明显差异帅>0.05人 三、血清垂体——甲状腺激素测定结果 二.冗 75天* 天各处理组大鼠 ’IT 7k平与对照组比较无明显差 异h>0.05入碘氟过量各组与碘过量组比较无明显差异h>0. 05人 150天氟过量 K、皿组几水平分别比同浓度 75天水平明 显降低(p<0.05)。 2。h 75天时各对照组 h水平与对照组比较无明显差异仲>0. 05入150天碘过量组 rIT水平与对照组比较无明显差异h>0· 05入碘氟过量各组兀水平与对照组比较无明显差异帅>0.0幻, 150天氟过量各组 ’IT水平与对照组比较明显降低帅<0.05X较 其 75天水平亦明显降低(p<0.05)。 3.TSH 75天*50天各处理组大鼠hH水平与对照组比较无明显差 异h>0.05X碘氟过量各组与碘过量比较亦无明显差异h>0. 05h碘过量组 150天水平与其 75天比较无显著差异讣>0.05八 4.甲状腺?

Preface: Iodine and fluoride are two essential trace elements for the human body, and the intake of iodine and fluoride by the body depends mainly on the external environment. Three years after the implementation of the Universal Salt Iodization (USI) regulation, our research group conducted an epidemiological investigation on thyroid diseases in rural communities in Liaoning and Hebei provinces with different iodine intake levels (median urinary iodine levels of 103μg/L, 374μg/L, and 615μg/L, respectively). The results showed that an increase in iodine intake led to a significant increase in hypothyroidism, autoimmune thyroiditis, and the incidence of papillary thyroid cancer. At the same time, we found that there were factors of high fluoride in high iodine areas. Excessive fluoride can cause varying degrees of metabolic disorders and toxic manifestations in the body, such as dental fluorosis and skeletal fluorosis. The impact of fluoride excess on the thyroid has been a long-standing debate among international scholars, and epidemiological investigations and animal experimental results on this matter are inconsistent. The mechanism of fluoride's impact on the thyroid is still unclear. This experiment aims to explore the effects of fluoride excess and combined iodine and fluoride excess on the thyroid through the measurement of iodine and fluoride metabolism in rats, thyroid hormone levels, and morphological observations. This will clarify the role of fluoride in thyroid damage, better understand the complex relationship between trace elements, and provide a theoretical basis for preventive measures for human health.

Experimental Materials:

Experimental Animals: 160 Wistar rats at the age of 5 weeks (80 males and 80 females) with a weight range of 125–165g were randomly divided into 8 groups. They were fed with regular feed and deionized water during the experiment.
Main Instruments: Electronic balance, automatic weight scale, stainless steel metabolic cages, paraffin embedding machine, water bath pot, incubator, pH meter, -20°C freezer, -70°C freezer, paraffin slicing machine, centrifuge, transmission electron microscope, optical microscope, microscopic image analysis system, universal microscope.
Experimental Methods:
a. Animal grouping and treatment: The 160 rats at 5 weeks of age were randomly divided into 8 groups, with 20 rats in each group, half male and half female. The animals were given different concentrations of sodium fluoride in deionized water freely, and the iodine excess group was given the same concentration using potassium iodate. The groups were as follows:
Control Group: Deionized water
Fluoride Excess Group 1: Deionized water containing 15ppm fluoride (equivalent to 3 times higher fluoride)
Fluoride Excess Group 2: Deionized water containing 30ppm fluoride (equivalent to 6 times higher fluoride)
Fluoride Excess Group 3: Deionized water containing 60ppm fluoride (equivalent to 12 times higher fluoride)
Iodine Excess Group: Deionized water containing 1200μg/L iodine (equivalent to 6 times higher iodine)
Iodine-Fluoride Excess Group 1: Deionized water containing 1200μg/L iodine and 15ppm fluoride (equivalent to 6 times higher iodine and 3 times higher fluoride)
Iodine-Fluoride Excess Group 2: Deionized water containing 1200μg/L iodine and 30ppm fluoride (equivalent to 6 times higher iodine and 6 times higher fluoride)
Iodine-Fluoride Excess Group 3: Deionized water containing 1200μg/L iodine and 60ppm fluoride (equivalent to 6 times higher iodine and 12 times higher fluoride)
b. Early euthanasia of half of the animals in each group at 75 days, with 10 rats per group, and the remaining rats euthanized at 150 days.
Specimen Collection and Processing:
Urine
Body weight
Serum
Thyroid and its wet weight
Conventional histological observation of tissues
The process of making paraffin sections: Fixation, dehydration, transparency, wax impregnation, embedding, and sectioning. (2) HE staining: Dewaxing, hydration, staining with hematoxylin and eosin, differentiation with 2% hydrochloric acid alcohol, bluing, counterstaining with eosin, washing, dehydration, transparency, and mounting.
Transmission electron microscope ultrastructural observation
Serum pituitary-thyroid hormone level measurement: T3, T4, and TSH levels were measured using radioimmunoassay (RIA) and solid-phase radioimmunoassay (IRMA).
Urinary iodine and fluoride determination: Arsenic-catalyzed spectrophotometry was used for urinary iodine, and fluoride ion-selective electrode method was used for urinary fluoride.
Image acquisition and analysis.
Data statistical processing: Single-factor analysis of variance and SNK test were performed using SPSS 10.0 software, with P<0.05 considered statistically significant.
Experimental Results:

Urinary iodine measurement results: The iodine-excess group and the iodine-fluoride-excess group showed a significant increase in urinary iodine levels compared to the control group at 75 days (P<0.05). There was no significant difference between the iodine-fluoride-excess groups and the iodine-excess group (P>0.05).
Urinary fluoride measurement results: The urinary fluoride levels of the fluoride-excess group and the iodine-fluoride-excess group were significantly higher than those of the control group at 75 days (P<0.05). The levels also increased significantly at 150 days compared to 75 days (P<0.05), and there was no significant difference between the iodine-fluoride-excess group and the fluoride-excess group (P>0.05).
Serum pituitary-thyroid hormone measurement results:
T4 levels in the fluoride-excess group at 75 days were significantly lower than those in the control group (P<0.05), and the levels in the fluoride-excess group at 150 days were significantly lower than those at 75 days (P<0.05).
T3 levels showed no significant difference between the control group and the fluoride-excess groups at 75 and 150 days (P>0.05).
TSH levels showed no significant difference between the control group and the fluoride-excess groups at 75 and 150 days (P>0.05).
Conclusion: The experimental results indicate that excess fluoride has a significant impact on the thyroid function of rats, affecting the metabolism of iodine and thyroid hormones. This study provides a theoretical basis for understanding the complex relationship between trace elements and their effects on human health, and it contributes to the development of preventive measures.
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