發(fā)布時間：2018-03-20 15:30

  本文選題：氟　切入點：亞硫酸鈉　出處：《山西農(nóng)業(yè)大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：[目的]研究氟和亞硫酸鈉單獨及聯(lián)合作用對小鼠睪丸組織中MAPK信號通路的影響,以探討氟和亞硫酸鈉對雄性生殖系統(tǒng)的毒性機制,為氟中毒防治以及亞硫酸鈉對雄性生殖系統(tǒng)的損害提供動物實驗依據(jù)。 [方法]模擬環(huán)境污染,以健康的雄性昆明小鼠為實驗動物,建立不同染毒時間(7周,14周)的小鼠實驗?zāi)Ｐ?設(shè)1個對照組和3個實驗組,整個試驗期間對照組小鼠自由飲用去離子水；氟組飲用150mg/L氟化鈉(NaF)去離子水;亞硫酸鈉組飲用500mg/L亞硫酸鈉(Na2SO3)去離子水；聯(lián)合組飲用150mg/L NaF+500mg/L Na2SO3去離子水。實驗結(jié)束時取小鼠的睪丸組織,應(yīng)用Real-time PCR技術(shù)檢測睪丸組織MAPK信號通路中ERK1/2/5、JNK1/2/3、p38及JNK途徑下游轉(zhuǎn)錄因子C-Jun、JunD、Elk-1、ATF-2基因mRNA表達(dá)量的變化。 [結(jié)果](1)Real-time PCR結(jié)果顯示,染毒7周時,與對照組相比,各實驗組ERK2基因mRNA的表達(dá)分別下降29.4%、52.9%、54.4%,其中亞硫酸鈉組和聯(lián)合組差異顯著(P0.05)。與對照組相比,氟組、亞硫酸鈉組、聯(lián)合組中JNK2基因mRNA的表達(dá)水平分別下降29.6%、59.3%、56.8%,其中亞硫酸鈉組和聯(lián)合組差異極顯著(P0.01)。而與對照組相比,氟組、亞硫酸鈉組、聯(lián)合組ERK1、JNK1、JNK3、ERK5基因mRNA的表達(dá)水平差異無統(tǒng)計學(xué)意義。 (2)Real-time PCR結(jié)果顯示,染毒14周時,與對照組相比,氟組、亞硫酸鈉組、聯(lián)合組JNK1基因1mRNA表達(dá)水平分別下降39.1%、61.8%、25.5%,其中亞硫酸鈉組組差異顯著(P0.05)；與對照組相比,氟組和亞硫酸鈉組和聯(lián)合組JNK2基因mRNA表達(dá)水平分別下降48.9%、39.8%、37.5%,三個實驗組差異均極顯著(P0.01),而與對照組相比,各實驗組ERK1、ERK2、ERK5、JNK3和p38基因mRNA的表達(dá)差異無統(tǒng)計學(xué)意義。 (3)Real-time PCR結(jié)果顯示,染毒7周時,氟組、亞硫酸鈉組、聯(lián)合組C-Jun、JunD、 Elk-1、ATF-2基因mRNA表達(dá)水平與對照組相比均沒有表現(xiàn)出顯著的差異；染毒14周時,與對照組相比,氟組、亞硫酸鈉組、聯(lián)合組C-Jun基因nRNA表達(dá)水平分別升高44.7%、31.2%、30.7%,差異均顯著(P0.05)；JunD基因mRNA表達(dá)水平分別升高92.1%、36.6%、150.5%,其中氟組差異顯著(P0.05),聯(lián)合組差異超顯著(P0.001)；氟組和聯(lián)合組Elk-1基因mRNA表達(dá)水平分別升高53.5%、34.9%,差異均顯著(P0.05),亞硫酸鈉則組下降8.1%；與對照組相比,ATF-2基因表達(dá)水平只有氟組與對照組相比差異顯著(p0.05),其余各組相比對照組均不顯著。 [結(jié)論]本研究采用熒光定量PCR技術(shù)分別對小鼠睪丸組織ERK1/2/5、JNK1/2/3、p38基因mRNA及JNK下游轉(zhuǎn)錄因子C-Jun、JunD、E1K-1和ATF-2mRNA基因表達(dá)變化進(jìn)行研究,結(jié)果表明本實驗?zāi)Ｐ椭?氟和亞硫酸鈉在JNK信號通路mRNA水平上對小鼠睪丸組織產(chǎn)生影響,并對小鼠睪丸JNK下游基因表達(dá)有影響,氟和亞硫酸鈉對雄性生殖系統(tǒng)損傷時通過激活JNK通路并影響下游C-Jun、JunD和Elk-1轉(zhuǎn)錄因子的活性來調(diào)控的,并且二者聯(lián)合中毒的過程中氟起主要作用,亞硫酸鈉對雄性生殖系統(tǒng)的毒性不強或者與氟化鈉有拮抗作用。
[Abstract]:[objective] to study the effects of fluoride and sodium sulfite alone and in combination on MAPK signaling pathway in mouse testis in order to explore the toxic mechanism of fluoride and sodium sulfite on male reproductive system. To provide experimental evidence for the prevention and treatment of fluorosis and the damage of sodium sulfite to male reproductive system. [methods] the healthy male Kunming mice were used as experimental animals to simulate environmental pollution. The experimental models of mice with different exposure time of 7 weeks and 14 weeks were established, and one control group and three experimental groups were set up. During the whole trial, the control mice drank deionized water freely; the fluorine group drank 150 mg / L sodium fluoride (NAF) deionized water; the sodium sulfite group drank 500mg / L sodium sulfite sodium sulfate (Na2SO3) deionized water; The combined group drank 150mg / L NaF 500mg / L Na2SO3 deionized water. At the end of the experiment, the testicular tissues of mice were taken. The changes of ERK1 / 2 / 2 / 5 / 5 JNK1 / 2 / 3p38 and JNK downstream transcription factor C-Jun-JunDN-Elk-1ATF-2 gene mRNA expression were detected by Real-time PCR technique. [results] the results of Real-time PCR showed that after 7 weeks of exposure, the expression of ERK2 gene mRNA in each experimental group was reduced by 29.4% and 52.9%, respectively. The difference between sodium sulfite group and combined group was significant (P0.055.Compared with the control group, fluoride group, sodium sulfite group, sodium sulfite group, sodium sulfite group, sodium sulfite group, sodium sulfite group, sodium sulfite group. The expression level of JNK2 gene mRNA in the combined group was reduced by 29.6and 59.3and 56.8, respectively. The difference between the sodium sulfite group and the combined group was very significant (P 0.01). However, there was no significant difference in the expression of ERK1 JNK1 JNK3ERK5 gene mRNA between the fluoride group, the sodium sulfite group and the combined group compared with the control group. The results of Real-time PCR showed that the expression level of JNK1 gene 1 in fluorine group, sodium sulfite group and combination group decreased by 39.1% and 61.8% respectively at 14 weeks after exposure, especially in sodium sulfite group (P 0.05), and compared with control group (P < 0.05). The expression of JNK2 mRNA in fluoride group, sodium sulfite group and combined group was decreased by 48.9% and 39.8%, respectively. The difference among the three experimental groups was very significant (P 0.01). However, there was no significant difference in the expression of ERK1, ERK2, ERK5, JNK3 and p38 gene mRNA between the three experimental groups. The results of Real-time PCR showed that there was no significant difference in the mRNA expression levels of C-Jun-JunD, Elk-1na-ATF-2 gene between fluorine group, sodium sulfite group and combined group at 7 weeks after exposure, and at 14 weeks after exposure, there was no significant difference between fluorine group and sodium sulfite group, but no significant difference was found between fluorine group and sodium sulfite group at 14 weeks after exposure. The expression level of C-Jun gene nRNA in the combined group was increased by 44.7% and 31.20.2%, respectively, and the difference was significant (P 0.05). The mRNA expression level of JunD gene increased 92.1% and 36.66%, respectively. The difference was significant in fluorine group (P 0.05), and in the combination group (P < 0.05). The Elk-1 gene mRNA expression level in fluorine group and combined group was higher than that in control group (P < 0.05), and the Elk-1 gene mRNA expression level in fluorine group and combined group was significantly higher than that in control group (P < 0.05). The difference was significant (P 0.05), and that of sodium sulfite group was 8.1%, and the expression level of ATF-2 gene was only significantly higher in fluorine group than that in control group (P 0.05), but not in other groups. [conclusion] in this study, the changes of ERK1 / 2 / 5 JNK1 / 2 / 3p38 gene mRNA and JNK downstream transcription factor C-JunJunDNE1K-1 and ATF-2mRNA gene expression in mouse testis were studied by fluorescence quantitative PCR technique. The results showed that, Fluoride and sodium sulfite had an effect on the testis of mice at the mRNA level of JNK signaling pathway, and on the expression of genes downstream of JNK in the testis of mice. Fluoride and sodium sulfite are regulated by activating the JNK pathway and affecting the activity of C-Jun-JunD and Elk-1 transcription factors downstream during male reproductive system injury, and fluoride plays a major role in the process of combined poisoning. Sodium sulfite is not toxic to male reproductive system or antagonistic to sodium fluoride.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R114

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