氟和亚硫酸钠对小鼠睾丸MAPK信号通路影响的研究

发布时间：2018-03-20 15:30

本文选题：氟　切入点：亚硫酸钠　出处：《山西农业大学》2013年硕士论文　论文类型：学位论文

【摘要】：[目的]研究氟和亚硫酸钠单独及联合作用对小鼠睾丸组织中MAPK信号通路的影响,以探讨氟和亚硫酸钠对雄性生殖系统的毒性机制,为氟中毒防治以及亚硫酸钠对雄性生殖系统的损害提供动物实验依据。 [方法]模拟环境污染,以健康的雄性昆明小鼠为实验动物,建立不同染毒时间(7周,14周)的小鼠实验模型,设1个对照组和3个实验组,整个试验期间对照组小鼠自由饮用去离子水；氟组饮用150mg/L氟化钠(NaF)去离子水;亚硫酸钠组饮用500mg/L亚硫酸钠(Na2SO3)去离子水；联合组饮用150mg/L NaF+500mg/L Na2SO3去离子水。实验结束时取小鼠的睾丸组织,应用Real-time PCR技术检测睾丸组织MAPK信号通路中ERK1/2/5、JNK1/2/3、p38及JNK途径下游转录因子C-Jun、JunD、Elk-1、ATF-2基因mRNA表达量的变化。 [结果](1)Real-time PCR结果显示,染毒7周时,与对照组相比,各实验组ERK2基因mRNA的表达分别下降29.4%、52.9%、54.4%,其中亚硫酸钠组和联合组差异显著(P0.05)。与对照组相比,氟组、亚硫酸钠组、联合组中JNK2基因mRNA的表达水平分别下降29.6%、59.3%、56.8%,其中亚硫酸钠组和联合组差异极显著(P0.01)。而与对照组相比,氟组、亚硫酸钠组、联合组ERK1、JNK1、JNK3、ERK5基因mRNA的表达水平差异无统计学意义。 (2)Real-time PCR结果显示,染毒14周时,与对照组相比,氟组、亚硫酸钠组、联合组JNK1基因1mRNA表达水平分别下降39.1%、61.8%、25.5%,其中亚硫酸钠组组差异显著(P0.05)；与对照组相比,氟组和亚硫酸钠组和联合组JNK2基因mRNA表达水平分别下降48.9%、39.8%、37.5%,三个实验组差异均极显著(P0.01),而与对照组相比,各实验组ERK1、ERK2、ERK5、JNK3和p38基因mRNA的表达差异无统计学意义。 (3)Real-time PCR结果显示,染毒7周时,氟组、亚硫酸钠组、联合组C-Jun、JunD、 Elk-1、ATF-2基因mRNA表达水平与对照组相比均没有表现出显著的差异；染毒14周时,与对照组相比,氟组、亚硫酸钠组、联合组C-Jun基因nRNA表达水平分别升高44.7%、31.2%、30.7%,差异均显著(P0.05)；JunD基因mRNA表达水平分别升高92.1%、36.6%、150.5%,其中氟组差异显著(P0.05),联合组差异超显著(P0.001)；氟组和联合组Elk-1基因mRNA表达水平分别升高53.5%、34.9%,差异均显著(P0.05),亚硫酸钠则组下降8.1%；与对照组相比,ATF-2基因表达水平只有氟组与对照组相比差异显著(p0.05),其余各组相比对照组均不显著。 [结论]本研究采用荧光定量PCR技术分别对小鼠睾丸组织ERK1/2/5、JNK1/2/3、p38基因mRNA及JNK下游转录因子C-Jun、JunD、E1K-1和ATF-2mRNA基因表达变化进行研究,结果表明本实验模型中,氟和亚硫酸钠在JNK信号通路mRNA水平上对小鼠睾丸组织产生影响,并对小鼠睾丸JNK下游基因表达有影响,氟和亚硫酸钠对雄性生殖系统损伤时通过激活JNK通路并影响下游C-Jun、JunD和Elk-1转录因子的活性来调控的,并且二者联合中毒的过程中氟起主要作用,亚硫酸钠对雄性生殖系统的毒性不强或者与氟化钠有拮抗作用。
[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.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

【参考文献】