本文選題：三氧化二砷　切入點(diǎn)：凋亡　出處：《大連醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討砷誘導(dǎo)小鼠海馬神經(jīng)元凋亡與Bcl-2和Bax表達(dá)之間的關(guān)聯(lián)性，為闡明砷的神經(jīng)毒作用機(jī)制以及預(yù)防神經(jīng)毒性危害提供依據(jù)。 方法：SPF級小鼠40只，雌雄各半，按體重將小鼠隨機(jī)分為4組，分別為對照組、1ppm As2O3染毒組、2ppm As2O3染毒組、4ppm As2O3染毒組。通過自然飲用含不同濃度As2O3蒸餾水的方式使小鼠染毒，連續(xù)染毒60天后取出海馬組織。用TUNEL染色法和電鏡技術(shù)檢測染砷小鼠海馬神經(jīng)元的凋亡情況，用Real-timePCR，Western blot技術(shù)檢測Bcl-2和Bax的基因、蛋白表達(dá)。采用SPSS11.5統(tǒng)計(jì)軟件，用單因素方差分析（ANOVA），比較各染砷組與對照組間的統(tǒng)計(jì)學(xué)差異，兩組間比較用LSD法分析，以P0.05表示統(tǒng)計(jì)學(xué)差異顯著。 結(jié)果：TUNEL凋亡染色結(jié)果顯示，對照組小鼠海馬組織CA1區(qū)未見陽性染色細(xì)胞，而染砷組小鼠海馬組織CA1區(qū)可見陽性染色細(xì)胞，且隨著染砷劑量的增加其陽性染色細(xì)胞數(shù)也增加，尤其2ppm染砷組4ppm染砷組小鼠海馬組織陽性細(xì)胞數(shù)量顯著多于對照組。電鏡檢測結(jié)果顯示，，染砷組小鼠海馬組織在視野中可見較多數(shù)量的凋亡細(xì)胞，細(xì)胞皺縮，染色質(zhì)凝聚分布于核周，與TUNEL凋亡染色結(jié)果基本一致。Real Time PCR定量檢測結(jié)果顯示，染砷組小鼠海馬組織中Bcl-2基因表達(dá)顯著低于對照組，而Bax基因表達(dá)顯著高于對照組(P0.05)。Western blot檢測結(jié)果顯示，染砷組的小鼠海馬組織中Bcl-2基因表達(dá)顯著低于對照組，而Bax基因表達(dá)顯著高于對照組(P0.05)，與Real-time PCR結(jié)果一致。 結(jié)論：亞慢性砷暴露誘導(dǎo)小鼠海馬神經(jīng)元凋亡，下調(diào)小鼠海馬組織Bcl-2基因、蛋白表達(dá)和上調(diào)小鼠海馬組織Bax基因、蛋白表達(dá)。亞慢性砷暴露小鼠海馬神經(jīng)元出現(xiàn)凋亡可能與砷導(dǎo)致的Bcl-2表達(dá)下調(diào)和Bax表達(dá)上調(diào)有關(guān)。
[Abstract]:Aim: to investigate the relationship between arsenic induced apoptosis of hippocampal neurons and the expression of Bcl-2 and Bax in mice, and to provide evidence for elucidating the neurotoxic mechanism of arsenic and preventing neurotoxicity. Methods Forty SPF mice, half male and half female, were randomly divided into 4 groups according to their body weight. The mice were exposed to 4 ppm As2O3 in the control group treated with 1 ppm As2O3 and 2 ppm As2O3. The mice were poisoned by drinking distilled water containing different concentrations of As2O3 naturally. The hippocampal tissue was taken out after 60 days of continuous exposure. Apoptosis of hippocampal neurons in arsenic exposed mice was detected by TUNEL staining and electron microscopy. The gene and protein expression of Bcl-2 and Bax were detected by Real-time PCR Western blot technique, and the expression of Bcl-2 and Bax were detected by SPSS11.5 statistical software. Single factor ANOVAX was used to compare the statistical difference between the arsenic exposed group and the control group, and the LSD method was used to analyze the difference between the two groups. Results the results of apoptosis staining showed that there were no positive staining cells in the CA1 region of hippocampal tissue in the control group, but there were positive staining cells in the CA1 area of the hippocampal tissue in the arsenic exposed group, and the number of positive staining cells increased with the increase of arsenic exposure dose. In particular, the number of positive cells in the hippocampus of 4ppm arsenic exposed mice in 2 ppm arsenic group was significantly higher than that in the control group. The results of electron microscopy showed that a large number of apoptotic cells were found in the hippocampal tissues of arsenic exposed mice, and the cells shrank. Chromatin condensed around the nucleus and was consistent with the results of TUNEL apoptosis staining. Real Time PCR quantitative analysis showed that the expression of Bcl-2 gene in the hippocampus of arsenic exposed mice was significantly lower than that of the control group. However, the expression of Bax gene was significantly higher than that of control group (P 0.05). Western blot analysis showed that the expression of Bcl-2 gene in hippocampus of arsenic exposed mice was significantly lower than that of control group, while the expression of Bax gene was significantly higher than that of control group (P 0.05), which was consistent with the result of Real-time PCR. Conclusion: subchronic arsenic exposure induces apoptosis of hippocampal neurons in mice, down-regulates Bcl-2 gene expression and up-regulates Bax gene expression in hippocampal tissues of mice. The apoptosis of hippocampal neurons in mice exposed to subchronic arsenic may be related to the down-regulation of Bcl-2 expression and the up-regulation of Bax expression induced by arsenic.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R114

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