镉暴露对大鼠性腺组织及基因甲基化的影响

发布时间：2018-11-05 13:16

【摘要】：为研究镉对SD大鼠性腺Bcl-2和Caspase-9基因甲基化水平的影响,将104只健康清洁级SD雄性大鼠随机分为A、B、C、D组,每组26只,对照组每天腹腔注射生理盐水,试验组每天分别用含0、0.3、0.6和1.2 mg/kg剂量的Cd Cl2溶液腹腔注射大鼠,期间观察、记录大鼠活动情况及体重变化,喂养28天后,取性腺、肝脏、肌肉、肾脏组织,测定组织中镉含量、SOD、CAT、GPX酶活性及MDA含量等相关指标,取性腺、肝脏组织行病理切片,HE染色。提取性腺、肝脏、肌肉、肾脏组织RNA,采用实时荧光定量RT-PCR法检测bcl-2、caspase-9、Caspase-3、BAX、AIF和CYCS基因在大鼠各组织中m RNA的表达量变化。提取性腺组织DNA,采用MSP法检测检测模型组大鼠性腺组织中Bcl-2和Caspase-9基因甲基化变化。采用Mass Array甲基化检测Bcl-2和Caspase-9基因CPG岛甲基化程度。用单因素方差分析及卡方检验对统计数据进行分析。1.试验组大鼠的体重增长速度慢于对照组,具有显著性差异(P0.05)。2.试验组大鼠性腺和肝脏组织富积镉的量均显著高于对照组,具有显著性差异(P0.05)。3.随着镉剂量的增大,与对照组相比,实验组大鼠各组织中MDA含量升高,GPX、SOD、CAT活性都有不同程度的先上升后下降的趋势,试验组与对照组差异显著或极显著(P0.05或P0.01)4.病理组织切片结果显示,试验组大鼠肝脏和性腺组织均出现不同程度的损伤。5.bcl-2基因甲基化水平受镉浓度的影响而明显增高,实时荧光定量表达量显著降低。而caspase-9基因甲基化水平受镉浓度影响不大,但比对照组有所升高,同时实时荧光定量表达量随着镉浓度升高显著增高。
[Abstract]:In order to study the effect of cadmium on the methylation of Bcl-2 and Caspase-9 gene in gonad of SD rats, 104 healthy and clean grade SD male rats were randomly divided into two groups: group A (n = 26) and control group (n = 26). Rats in the experimental group were injected intraperitoneally with Cd Cl2 solution containing 0 0. 3U 0. 6 and 1. 2 mg/kg each day. The rats were observed and recorded. After 28 days of feeding, the gonad, liver, muscle and kidney were taken out. The content of cadmium, the activity of SOD,CAT,GPX enzyme and the content of MDA were measured. The gonads and liver tissues were stained with HE and pathological sections. RNA, was extracted from gonad, liver, muscle and kidney to detect the expression of m RNA in rat tissues by real-time fluorescence quantitative RT-PCR. The methylation of Bcl-2 and Caspase-9 genes in the gonad tissue of the model group was detected by MSP method. Bcl-2 and Caspase-9 gene CPG island methylation was detected by Mass Array methylation. Single factor ANOVA and chi-square test were used to analyze the statistical data. 1. The weight gain rate of the experimental group was slower than that of the control group, with significant difference (P0.05). The amount of cadmium rich in gonads and liver tissues in the experimental group was significantly higher than that in the control group (P0.05). With the increase of cadmium dose, compared with the control group, the content of MDA in each tissue of the experimental group increased, and the activity of GPX,SOD,CAT increased at first and then decreased. The difference between the test group and the control group was significant or extremely significant (P0.05 or P0.01) 4. The results of histopathological sections showed that the liver and gonad tissues in the experimental group were damaged to varying degrees, the methylation level of 5.bcl-2 gene was significantly increased by the concentration of cadmium, and the quantitative expression of real-time fluorescence was significantly decreased. However, the methylation level of caspase-9 gene was not affected by cadmium concentration, but it was higher than that of control group, and the real-time fluorescence quantitative expression increased significantly with the increase of cadmium concentration.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X503.22

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