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青春期氰戊菊酯暴露對小鼠海馬甾體激素合成酶及雄激素受體和雌激素受體的影響

發(fā)布時間:2019-03-12 18:42
【摘要】:目的本課題通過觀察青春期暴露氰戊菊酯對小鼠大腦海馬甾體激素合成關(guān)鍵酶(stAR, P450scc, CYP17A1,17β-HSD,3β-HSD和CYP19)及甾體激素受體(AR,ERα和ERβ)表達(dá)的調(diào)控作用,闡明青春期暴露氰戊菊酯產(chǎn)生內(nèi)分泌干擾效應(yīng)的分子機理。 方法實驗用ICR小鼠從北京維通利華實驗動物技術(shù)有限公司購買。購買出生后4周ICR小鼠72只(36只雄鼠和36只雌鼠)。體重18-25g左右,雌雄分開并用常規(guī)飼料喂食,按照性別與體重順序編號,隨機分籠。分為4組,分別為溶劑對照組、低劑量氰戊菊酯組、中劑量氰戊菊酯組、高劑量氰戊菊酯組,氰戊菊酯濃度分別為0.02mg/kg,0.2mg/kg,2.0mg/kg。實驗前動物自由進(jìn)食標(biāo)準(zhǔn)飼料,維持12小時光照和12小時黑夜的晝夜規(guī)律。實驗室溫度:20-25℃,濕度:50±5%。實驗動物適應(yīng)性飼養(yǎng)1周后,開始按照先前的分組給予灌胃給藥,每天按照小鼠體重的1%的量給藥一次。每周稱一次體重,連續(xù)灌胃一個月后,剖殺老鼠。所有實驗均需得到安徽醫(yī)科大學(xué)醫(yī)學(xué)倫理委員會批準(zhǔn)。取海馬用于下列實驗:部分海馬組織用液氮速凍后-80℃保存,用于實時定量RT-PCR實驗;部分海馬組織用液氮速凍后-80℃保存,用于蛋白免疫印跡實驗。 結(jié)果 1.小鼠青春期和成年期階段均無疾病、感染和死亡現(xiàn)象,體重的差異無統(tǒng)計學(xué)意義。 2.各劑量組雄鼠的17β-HSD, P450scc以及(0.2mg/kg,2mg/kg)劑量組AR的蛋白水平與溶劑對照組相比較均顯著下調(diào),,差異均有統(tǒng)計學(xué)意義。 3.雄性(0.2mg/kg,2mg/kg)劑量組小鼠AR, cyp17a1, cyp19, ERα, ERβ及各劑量組小鼠的p450scc, star的mRNA表達(dá)與溶劑對照組相比較均顯著下調(diào),差異均有統(tǒng)計學(xué)意義。雌性(0.02mg/kg)劑量組小鼠p450scc,ERα和(0.2mg/kg)劑量組小鼠star,cyp19,ERβ的mRNA表達(dá)均顯著上調(diào),雌性小鼠各劑量組AR的mRNA 表達(dá)均明顯下調(diào),差異均有統(tǒng)計學(xué)意義。結(jié)論小鼠青春期暴露氰戊菊酯干擾各劑量組甾體激素合成關(guān)鍵酶以及雄激素受體和雌激素受體的蛋白表達(dá)水平和mRNA表達(dá)水平,并存在明顯的性別差異。
[Abstract]:Objective to investigate the regulatory effects of fenvalerate on the expression of stAR, P450scc (CYP17A1,17 尾-HSD,3 尾-HSD and CYP19) and steroid hormone receptor (AR, ER 偽 and ER 尾) in hippocampus of mice exposed to fenvalerate during puberty. To elucidate the molecular mechanism of endocrine disruptive effect of fenvalerate during puberty exposure. Methods ICR mice were purchased from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. 72 ICR mice (36 males and 36 females) were purchased 4 weeks after birth. The weight was about 18g / 25g. The male and female were divided into two groups randomly according to the order of sex and body weight. They were divided into 4 groups: solvent control group, low-dose fenvalerate group, middle-dose fenvalerate group, high-dose fenvalerate group. The concentration of fenvalerate was 0.02 mg / kg, 0.2 mg / kg, 2.0 mg / kg, respectively. Before the experiment, animals were free to eat the standard diet, maintaining 12 hours of light and 12 hours of dark day and night. Laboratory temperature: 20 鈩

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