【摘要】：為研究鎘對SD大鼠性腺Bcl-2和Caspase-9基因甲基化水平的影響,將104只健康清潔級SD雄性大鼠隨機(jī)分為A、B、C、D組,每組26只,對照組每天腹腔注射生理鹽水,試驗組每天分別用含0、0.3、0.6和1.2 mg/kg劑量的Cd Cl2溶液腹腔注射大鼠,期間觀察、記錄大鼠活動情況及體重變化,喂養(yǎng)28天后,取性腺、肝臟、肌肉、腎臟組織,測定組織中鎘含量、SOD、CAT、GPX酶活性及MDA含量等相關(guān)指標(biāo),取性腺、肝臟組織行病理切片,HE染色。提取性腺、肝臟、肌肉、腎臟組織RNA,采用實時熒光定量RT-PCR法檢測bcl-2、caspase-9、Caspase-3、BAX、AIF和CYCS基因在大鼠各組織中m RNA的表達(dá)量變化。提取性腺組織DNA,采用MSP法檢測檢測模型組大鼠性腺組織中Bcl-2和Caspase-9基因甲基化變化。采用Mass Array甲基化檢測Bcl-2和Caspase-9基因CPG島甲基化程度。用單因素方差分析及卡方檢驗對統(tǒng)計數(shù)據(jù)進(jìn)行分析。1.試驗組大鼠的體重增長速度慢于對照組,具有顯著性差異(P0.05)。2.試驗組大鼠性腺和肝臟組織富積鎘的量均顯著高于對照組,具有顯著性差異(P0.05)。3.隨著鎘劑量的增大,與對照組相比,實驗組大鼠各組織中MDA含量升高,GPX、SOD、CAT活性都有不同程度的先上升后下降的趨勢,試驗組與對照組差異顯著或極顯著(P0.05或P0.01)4.病理組織切片結(jié)果顯示,試驗組大鼠肝臟和性腺組織均出現(xiàn)不同程度的損傷。5.bcl-2基因甲基化水平受鎘濃度的影響而明顯增高,實時熒光定量表達(dá)量顯著降低。而caspase-9基因甲基化水平受鎘濃度影響不大,但比對照組有所升高,同時實時熒光定量表達(dá)量隨著鎘濃度升高顯著增高。
[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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：X503.22

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