2015: Effects of fluoride on TH - Zebrafish

2015: Effects of fluoride on TH - Zebrafish

Postby admin » Tue Jun 23, 2020 11:22 pm

199. 薛文娟 氟对斑马鱼下丘脑—垂体—甲状腺轴干扰效应的研究 《山西农业大学》 2015年收藏 | 手机打开
http://cdmd.cnki.com.cn/Article/CDMD-10 ... 623693.htm
Xue Wenjuan
"Effect of fluoride on the zebrafish hypothalamic-pituitary-thyroid axis"

【摘要】:目前已有大量的研究表明高氟会抑制鱼类的生长发育,但对其影响机制还尚无定论。内分泌的调节对机体的生长发育至关重要,下丘脑-垂体-甲状腺(HPT)轴是内分泌系统的成员之一。为探究氟是否通过影响鱼类HPT轴影响鱼类正常生长发育,本试验选用两月龄健康斑马鱼,随机分为3组,分别为对照组(0mg/L),40mg/L和80 mg/L氟暴露组。分别在氟暴露45 d和90 d时,对斑马鱼的生长发育状况进行记录,并通过HE染色对斑马鱼甲状腺进行组织病理学观察,用酶联免疫ELISA检测鱼体内T3和T4激素水平,最后对斑马鱼头部和肝脏组织提取总RNA,应用实时荧光定量PCR方法检测HPT轴相关基因的表达。试验结果如下:1.对生长发育分析结果显示,氟暴露45 d时,雌雄斑马鱼体重、增重率、特定增重率及肥满度均表现为40 mg/L促进80 mg/L抑制,体长及增长率均表现为随着浓度的升高生长抑制作用越大。氟暴露90天时除雄鱼肥满度外,其他各性别各指标均随着浓度的上升而下降。2.甲状腺组织病理学结果显示,氟暴露可造成斑马鱼甲状腺滤泡的病变,包括滤泡肿大、上皮细胞呈扁平状、增生和内褶,胶质萎缩或呈泡沫状等。且随着暴露浓度增大和时间的延长,甲状腺滤泡的病变越严重;肝脏组织病理学结果显示,40 mg/L氟暴露组在45 d时就发生了脂肪样变、血栓等病变,且暴露浓度越高越严重。在氟暴露90d时,雌鱼肝脏脂肪样变、血栓情况加深,雄鱼80 mg/L组肝细胞大量溶解,细胞核聚集,肝组织严重受到损伤。3.甲状腺激素T3和T4 ELISA结果显示,氟暴露可影响斑马鱼体内甲状腺激素T3和T4的水平及T3/T4的比值,且影响程度随暴露时间、浓度及性别不同而不同。总体上分析,暴露45 d时,氟可提高T3和T4水平,但T3/T4比值降低。氟暴露90 d时,除80mg/L组雌鱼T3水平和T3/T4比值降低外,其他各组T3水平和T3/T4比值均升高,但T4水平均降低。4.对氟暴露后斑马鱼HPT轴CRH、TSH、TG、NIS, TTR、DIO1、DIO2、TRα、TRβ和iUGT1ab等10个基因的表达量结果分析显示:氟暴露45 d时,雌斑马鱼40 mg/L组除CRH与TRRα外,其他8个基因表达均显著下调,80 mg/L组除TG、NIS、TTR及TRα外,其他6个基因表达均显著下调;雄鱼氟暴露45 d时,40mg/L组除CRH, NIS及D1O1外,其他7个基因表达均显著上调,80 mg/L组除UGT1ab与TTR基因外,其他8个基因表达均显著上调。氟暴露90 d时,雌鱼40 mg/L组DIO1、DIO2 mRNA和80 mg/L组TG、DIO1、 DIO2、TRα、TRβ mRNA表达显著上调,40 mg/L组TTR、TRα和80 mg/L组UGT1ab mRNA表达显著下调,其余都未有显著变化;雄鱼40 mg/L组TSH、TG、DIO1、TTR、TRα和80mg/L TG、DIO1、TRα mRNA表达均显著上调,40mg/L组CRH和80mg/L组CRH、DIO2、TTR、UGT1ab mRNA表达显著下调。试验结果表明,氟暴露可通过影响斑马鱼甲状腺和肝脏组织形态、甲状腺激素水平影响其生长发育,且随着暴露浓度的增加及暴露时间的延长影响逐渐增加。氟对斑马鱼HPT轴10个相关基因的表达均有影响,且影响的大小与暴露浓度、时间和性别有关,提示氟可通过影响斑马鱼HPT轴中基因的表达来干扰机体内甲状腺激素的含量,进而影响机体的生长发育。本试验研究结果为氟影响斑马鱼生长发育机制提供基础依据。
【学位授予单位】:山西农业大学
【学位级别】:硕士
【学位授予年份】:2015
【分类号】:S917.4
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Postby admin » Tue Jun 23, 2020 11:23 pm

Effect of fluoride on the zebrafish hypothalamic-pituitary-thyroid axis interference
Xue Wenjuan
http://cdmd.cnki.com.cn/Article/CDMD-10 ... 623693.htm
【Summary】:A large number of studies have shown that high fluoride will inhibit the growth and development of fish, but the mechanism of its influence is still inconclusive. The regulation of endocrine is crucial to the growth and development of the body. The hypothalamic-pituitary-thyroid (HPT) axis is one of the members of the endocrine system. In order to investigate whether fluorine affects the normal growth and development of fish by affecting the HPT axis of the fish, two-month-old healthy zebrafish were selected in this experiment and randomly divided into 3 groups, namely the control group (0mg/L), 40mg/L and 80mg/ L fluoride exposure group. The growth and development of zebrafish were recorded at 45 and 90 days of exposure to fluoride, respectively, and histopathological observation of the thyroid of zebrafish was performed by HE staining. T3 and T4 hormone levels in fish were detected by enzyme-linked immunoELISA. Total RNA was extracted from zebrafish head and liver tissues, and the expression of HPT axis related genes was detected by real-time fluorescence quantitative PCR. The test results are as follows: 1. The results of growth and development analysis showed that after exposure to fluoride for 45 days, the body weight, weight gain rate, specific weight gain rate and fullness of male and female zebrafish showed that 40 mg/L promoted 80 mg/L inhibition, Both body length and growth rate show greater growth inhibition with increasing concentration. In addition to the fullness of male fish, the other indicators of all genders decreased with the increase of concentration at 90 days of fluorine exposure. 2. Thyroid histopathological results showed that fluoride exposure can cause zebrafish thyroid follicular lesions, including enlarged follicles, flattened epithelial cells, hyperplasia and internal folds, colloidal atrophy or foam. And with the increase of exposure concentration and the extension of time, the lesions of thyroid follicles became more serious; the histopathological results of liver showed that fat-like changes, thrombosis and other lesions occurred in the 40 mg/L fluoride exposure group at 45 days, and the exposure The higher the concentration, the worse. After exposure to fluoride for 90 days, the fatty fat and thrombosis in the liver of female fish deepened. The 80 mg/L group of male fish had a large amount of hepatocytes dissolved, nuclei aggregated, and liver tissues were seriously damaged. 3. Thyroid hormone T3 and T4 ELISA results show that fluoride exposure can affect the levels of thyroid hormone T3 and T4 and the ratio of T3/T4 in zebrafish, and the degree of influence varies with exposure time, concentration and gender. Overall analysis, when exposed for 45 days, fluorine can increase T3 and T4 levels, but the T3/T4 ratio decreases. After exposure to fluoride for 90 days, the T3 level and T3/T4 ratio of the females in the 80mg/L group decreased, while the T3 level and T3/T4 ratio of the other groups increased, but the T4 level decreased. 4. After exposure to fluoride, the zebrafish HPT axis CRH, TSH, TG, NIS, The analysis of the expression of 10 genes such as TTR, DIO1, DIO2, TRα, TRβ, and iUGT1ab showed that the expression of the other 8 genes in the female zebrafish 40 mg/L group except CRH and TRRα was significantly down-regulated when exposed to fluorine for 45 days. In the 80 mg/L group, except for TG, NIS, TTR, and TRα, the expression of 6 other genes was significantly down-regulated; when the male fish was exposed to fluoride for 45 days, the other 7 genes in the 40 mg/L group except CRH, NIS, and D1O1 were expressed. All of them were significantly up-regulated. In the 80 mg/L group, except for UGT1ab and TTR genes, the expressions of the other 8 genes were significantly up-regulated. When exposed to fluoride for 90 days, the expression of DIO1, DIO2 mRNA in the 40 mg/L group and TG, DIO1, DIO2, TRα, and TRβ mRNA in the 80 mg/L group were significantly increased, and the TTR, TRα, and 80 mg/L in the 40 mg/L group. UGT1ab mRNA expression was significantly down-regulated in the group, and the rest were not significantly changed; TSH, TG, DIO1, TTR, TRα and 80mg/L TG, DIO1, TRα mRNA expression were significantly up-regulated in the 40mg/L male fish group, CRH in the 40mg/L group And 80mg/L group CRH, DIO2, TTR, UGT1ab mRNA expression was significantly down-regulated. The test results show that fluoride exposure can affect the growth and development of zebrafish thyroid and liver tissues, thyroid hormone levels, and gradually increase with the increase of exposure concentration and exposure time. Fluoride has an effect on the expression of 10 related genes in the zebrafish HPT axis, and the size of the effect is related to exposure concentration, time and gender, suggesting that fluoride can interfere with the content of thyroid hormones in the body by affecting the expression of genes in the zebrafish HPT axis And then affect the growth and development of the body. The results of this experiment provide a basis for the mechanism of fluoride affecting the growth and development of zebrafish. The expression of TG, DIO1, DIO2, TRα, TRβ mRNA in female 40 mg/L group and the expression of TG, DIO1, DIO2, TRα, TRβ in 80 mg/L group were significantly up-regulated, while the expression of TTR, TRα in 40 mg/L group and UGT1ab mRNA in 80 mg/L group were significantly down-regulated. , The rest did not change significantly; male fish 40 mg/L group TSH, TG, DIO1, TTR, TRα and 80 mg/L TG, DIO1, TRα mRNA expression were significantly up-regulated, 40 mg/L group CRH and 80 mg/L group CRH , DIO2, TTR, UGT1ab mRNA expression was significantly down-regulated. The test results show that fluoride exposure can affect the growth and development of zebrafish thyroid and liver tissues, thyroid hormone levels, and gradually increase with the increase of exposure concentration and exposure time. Fluoride has an effect on the expression of 10 related genes in the zebrafish HPT axis, and the size of the effect is related to exposure concentration, time and gender, suggesting that fluoride can interfere with the content of thyroid hormones in the body by affecting the expression of genes in the zebrafish HPT axis And then affect the growth and development of the body. The results of this experiment provide a basis for the mechanism of fluoride affecting the growth and development of zebrafish. The expression of TG, DIO1, DIO2, TRα, TRβ mRNA in female 40 mg/L group and the expression of TG, DIO1, DIO2, TRα, TRβ in 80 mg/L group were significantly up-regulated, while the expression of TTR, TRα in 40 mg/L group and UGT1ab mRNA in 80 mg/L group were significantly down-regulated. , The rest did not change significantly; male fish 40 mg/L group TSH, TG, DIO1, TTR, TRα and 80 mg/L TG, DIO1, TRα mRNA expression were significantly up-regulated, 40 mg/L group CRH and 80 mg/L group CRH , DIO2, TTR, UGT1ab mRNA expression was significantly down-regulated. The test results show that fluoride exposure can affect the growth and development of zebrafish thyroid and liver tissues, thyroid hormone levels, and gradually increase with the increase of exposure concentration and exposure time. Fluoride has an effect on the expression of 10 related genes in the zebrafish HPT axis, and the size of the effect is related to exposure concentration, time and gender, suggesting that fluoride can interfere with the content of thyroid hormones in the body by affecting the expression of genes in the zebrafish HPT axis And then affect the growth and development of the body. The results of this experiment provide a basis for the mechanism of fluoride affecting the growth and development of zebrafish.
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