本文選題：BPA + 新生雌性SD大鼠��； 參考：《蘇州大學》2014年碩士論文

【摘要】：目的：研究新生期不同劑量雙酚A（bisphenol A，BPA）暴露對雌性SD大鼠陰道開口時間（vaginal opening day，VOD）、下丘腦kiss-1mRNA及其蛋白kisspeptin表達和子宮、卵巢雌激素受體α（Estrogen Receptor，ERα）mRNA、雌激素受體β（Estrogen Receptor，ERβ）mRNA及其蛋白表達的影響。 方法：將新生雌性SD大鼠隨機分為6組，即空白對照組、溶劑組、17β-雌二醇組（17β-estradiol，E2，10μg d-1）、低劑量BPA組（25μg kg-1d-1）、中劑量BPA組（50μg kg-1d-1）和高劑量BPA組（250μg kg-1d-1），在生后第0-6天（postnatal day0-6，PND0-6）經(jīng)皮下注射每日給藥。記錄VOD，并于當天處死大鼠，取出下丘腦、子宮、卵巢并稱重，計算下丘腦、子宮、卵巢臟器系數(shù)。對下丘腦組織行免疫組化檢測觀察kisspeptin定位，子宮、卵巢行HE染色觀察形態(tài)變化。通過Real—Time PCR的方法測定下丘腦kiss-1mRNA和子宮、卵巢中ERα、ERβ mRNA表達量。通過Western-blot的方法測定下丘腦中kisspeptin及子宮、卵巢中ERα、ERβ蛋白表達量。 結(jié)果：1.對體重的影響：自PND0至PND6每日對大鼠進行皮下注射方式的藥物干預，對照組與各干預組間大鼠PND0與PND6體重未有明顯差異（P0.05）。2. VOD記錄顯示與對照組相比，E2組、50μg kg-1d-1和250μg kg-1d-1BPA組VOD較對照組提前（P0.05）；溶劑組、25μg kg-1d-1BPA組VOD與對照組相比未有顯著差異（P0.05）。3.下丘腦免疫組化結(jié)果提示kisspeptin多位于神經(jīng)細胞上。4. HE染色結(jié)果顯示空白對照組、溶劑組、25μg kg-1d-1、50μg kg-1d-1和250μg kg-1d-1BPA組子宮肌層厚度、內(nèi)膜厚度較E2組厚，卵巢卵泡發(fā)育度較E2組高。5.臟器系數(shù)結(jié)果顯示，各干預組下丘腦臟器系數(shù)與對照組相比無顯著差異（P0.05），但E2組子宮、卵巢臟器系數(shù)較對照組明顯降低（P0.05），溶劑組、BPA干預組較對照組無明顯改變（P0.05）。6. RT-PCR結(jié)果顯示，，溶劑組、25μg kg-1d-1、、50μg kg-1d-1和250μg kg-1d-1BPA組下丘腦kiss-1mRNA及子宮、卵巢ER mRNA表達水平較對照組未有明顯差異（P0.05）； E2組下丘腦kiss-1、子宮ERα、卵巢ERβ mRNA表達水平較對照組明顯降低（P0.05），子宮ERβ、卵巢ERα mRNA水平較對照組未有明顯差異（P0.05）。7. Western-Blot結(jié)果顯示，溶劑組、25μg kg-1d-1、50μg kg-1d-1和250μg kg-1d-1BPA組下丘腦kisspeptin及子宮、卵巢ER蛋白水平較對照組未有明顯差異（P0.05）； E2組下丘腦kisspeptin、子宮ERα、卵巢ERβ蛋白水平較對照組明顯降低（P0.05），子宮ERβ、卵巢ERα蛋白水平較對照組未有明顯差異（P0.05）。 結(jié)論：①每日皮下注射操作未對注射藥物期間大鼠基本體重增長產(chǎn)生明顯影響。②下丘腦神經(jīng)細胞上有kisspeptin表達。③新生期50μg kg-1d-1、250μg kg-1d-1BPA暴露可引起大鼠青春期啟動提前。④上述青春期啟動提前并沒有通過kiss-1基因表達改變來實現(xiàn)，且未對其青春期啟動時下丘腦、子宮、卵巢臟器系數(shù)，子宮、卵巢發(fā)育形態(tài)及ER表達產(chǎn)生明顯影響。⑤新生期溶劑及25μg kg-1d-1BPA暴露未引起大鼠青春期啟動提前。⑥新生期10μg d-1E2暴露可引起大鼠青春期啟動提前，且啟動時子宮、卵巢發(fā)育程度降低，子宮、卵巢臟器系數(shù)、下丘腦kiss-1mRNA、子宮ERα mRNA、卵巢ERβ mRNA表達量及其蛋白均較對照組明顯降低。
[Abstract]:Objective: To study the vaginal opening time (vaginal opening day, VOD) of female SD rats exposed to different doses of bisphenol A (bisphenol A, BPA), and the kisspeptin expression of kiss-1mRNA and its protein in the hypothalamus and the uterus, ovarian estrogen receptor alpha, estrogen receptor beta and its protein expression. Influence.
Methods: the newborn female SD rats were randomly divided into 6 groups, namely the blank control group, the solvent group, the 17 beta estradiol group (17 beta -estradiol, E2,10 mu g D-1), the low dose BPA group (25 mu g kg-1d-1), the medium dose BPA group (50 mu g kg-1d-1) and the high dose BPA group (250 mu), and the daily delivery was recorded by subcutaneous injection on the 0-6 day after birth. OD, the rats were killed on the same day, and the hypothalamus, uterus, ovary and weight were taken out. The hypothalamus, uterus, and ovarian organ coefficient were calculated. The hypothalamus was detected by immunohistochemical staining, and the morphology of the uterus and ovary were observed by HE staining. The kiss-1mRNA and uterus in the hypothalamus, ER a, ER beta m in the ovary were measured by Real Time PCR. The expression level of RNA was measured by Western-blot. The expression level of ER and ER beta in kisspeptin and uterus and ovary of hypothalamus were measured.
Results: the effect of 1. on weight: the intervention of subcutaneous injection of subcutaneous injection from PND0 to PND6, and there was no significant difference in the weight of PND0 and PND6 between the control group and the intervention group (P0.05).2. VOD record. Compared with the control group, the group E2, the 50 micron kg-1d-1 and the 250 mu g kg-1d-1BPA group were earlier than the control group; the solvent group, 25 There was no significant difference in VOD between the G kg-1d-1BPA group and the control group (P0.05) the results of the immunohistochemical staining of the hypothalamus in.3. showed that kisspeptin was mostly located on the nerve cells and the.4. HE staining results showed the blank control group, the solvent group, the 25 micron kg-1d-1,50 mu g kg-1d-1 and the 250 Mu uterine myometrium thickness, the thickness of the endometrium was thicker than that of the ovarian follicle development. The higher.5. organ coefficient compared with the E2 group showed that the hypothalamic organ coefficient of the intervention group had no significant difference compared with the control group (P0.05), but the uterus, the ovarian organ coefficient of the E2 group was significantly lower than the control group (P0.05), the solvent group and the BPA intervention group had no significant changes (P0.05).6. RT-PCR results (P0.05), the solvent group, 25 mu g kg-1d-1, 50 micron g, and 2. The expression level of ER mRNA in the hypothalamus of the 50 g kg-1d-1BPA group was not significantly different than that of the control group (P0.05). The level of KiSS-1 in the hypothalamus, ER alpha and ER beta in the E2 group was significantly lower than that of the control group (P0.05), and there was no significant difference between the uterus and the ovary of the control group. The level of kisspeptin and uterus of the hypothalamus in the group of solvent, 25 g kg-1d-1,50 g kg-1d-1 and 250 mu g kg-1d-1BPA group was not significantly different from that of the control group (P0.05), and the kisspeptin of the hypothalamus in the E2 group, the uterus ER alpha and the ovarian beta protein level were significantly lower than those in the control group. Significant difference (P0.05).
Conclusion: (1) daily subcutaneous injection did not significantly affect the basic weight growth of rats during the period of injection. (2) there was kisspeptin expression on the hypothalamus nerve cells. (3) 50 g kg-1d-1250 mu g kg-1d-1BPA exposure in the newborn period could lead to early puberty initiation in rats. (4) the early puberty start up did not pass through the KiSS-1 gene expression. Change to achieve, and did not have a significant effect on the hypothalamus, uterus, ovarian organ coefficient, uterus, ovarian development morphology and ER expression during puberty. 5. The neonatal period solvent and 25 g kg-1d-1BPA exposure did not lead to early puberty initiation. The degree of uterine and ovarian development decreased, the uterus, the ovarian organ coefficient, the kiss-1mRNA of the hypothalamus, the ER alpha mRNA of the uterus, the expression of ER beta mRNA in the ovary and their proteins were significantly lower than those of the control group.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R714.2

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