本文選題：亞砷酸鈉　切入點(diǎn)：大鼠胰島細(xì)胞　出處：《大連醫(yī)科大學(xué)》2012年碩士論文

【摘要】：前言：砷（As）是自然界中普遍存在的一種毒性很強(qiáng)的類金屬元素。慢性砷中毒已成為全球面臨的重大公共衛(wèi)生問題，我國(guó)有很多慢性砷中毒病區(qū)。為了保障人群健康,世界衛(wèi)生組織（WHO）、歐盟、日本和美國(guó)等先后將飲用水中砷的標(biāo)準(zhǔn)修訂為10μg/L，我國(guó)新的《生活飲用水衛(wèi)生標(biāo)準(zhǔn)》（2006年7月1日實(shí)施）也將砷含量標(biāo)準(zhǔn)由50μg/L降低到10μg/L。 國(guó)際癌癥研究機(jī)構(gòu)（IARC）和美國(guó)疾病控制中心（CDC）已確定砷是第一類致癌物質(zhì)。已有研究表明：無機(jī)砷可引起皮膚癌和肺癌，并且與肝癌、膀胱癌、腎臟癌、大腸癌的發(fā)生也密切相關(guān)。此外，有研究發(fā)現(xiàn)顯示亞砷酸鈉（NaAsO_2）具有遺傳毒性，但有關(guān)遺傳毒性機(jī)制的研究不多。研究表明，NaAsO_2可導(dǎo)致細(xì)胞中活性氧類（ROS）生成增多和8-羥基脫氧鳥苷（8-OHdG）表達(dá)升高。 本研究通過觀察不同濃度NaAsO_2對(duì)大鼠胰島β細(xì)胞株INS-1內(nèi)ROS和谷胱甘肽（GSH）水平、8-OHdG表達(dá)和DNA損傷的影響的影響，從氧化應(yīng)激角度探討砷所導(dǎo)致細(xì)胞DNA損傷的可能機(jī)制，為進(jìn)一步的深入研究砷所引起的機(jī)體氧化損傷提供基礎(chǔ)資料。 方法：以INS-1細(xì)胞作為實(shí)驗(yàn)對(duì)象，通過單細(xì)胞凝膠電泳（SCGE）試驗(yàn)檢測(cè)細(xì)胞DNA損傷情況，，評(píng)價(jià)NaAsO_2的遺傳毒性：為探討砷的遺傳毒性機(jī)制，以2’,7’-二氫二氯熒光素（DCFH-DA）法和苯二醛（OPT）分別測(cè)定細(xì)胞內(nèi)ROS以及GSH水平，以免疫組化方法檢測(cè)細(xì)胞內(nèi)8-OHdG的表達(dá)水平。實(shí)驗(yàn)結(jié)果用SPSS11.5統(tǒng)計(jì)。 結(jié)果：NaAsO_2（2-16μmol/L）作用于INS-1細(xì)胞12h后，可引起DNA鏈斷裂，細(xì)胞形成彗星樣拖尾，拖尾細(xì)胞數(shù)明顯增多，且呈劑量依賴關(guān)系。NaAsO_2（2-16μmol/L）作用于INS-1細(xì)胞12h后，可致8-OHdG的表達(dá)升高；NaAsO_2（2-16μmol/L）分別作用于INS-1細(xì)胞6h和12h后，可致細(xì)胞內(nèi)ROS表達(dá)水平的顯著增加，且呈劑量依賴關(guān)系，尤其是作用12h后，細(xì)胞內(nèi)ROS表達(dá)水平最為顯著；8μmol/L和16μmol/L NaAsO_2作用于INS-1細(xì)胞24h后，細(xì)胞內(nèi)ROS表達(dá)水平增加；NaAsO_2（2-16μmol/L）分別作用于INS-1細(xì)胞48h后，細(xì)胞內(nèi)ROS表達(dá)水平未見明顯改變。NaAsO_2（2-16μmol/L）作用于INS-1細(xì)胞6，12，24和48h后，可致細(xì)胞內(nèi)GSH表達(dá)水平的顯著增加，且呈劑量依賴關(guān)系。 結(jié)論：NaAsO_2通過氧化應(yīng)激機(jī)制引起INS-1細(xì)胞DNA損傷。
[Abstract]:The arsenic (As) is a kind of toxic metal elements exist in nature is very strong. Chronic arsenic poisoning has become a major public health problem facing the world, our country has a lot of chronic arsenic poisoning area. In order to protect people's health, the WHO (WHO), the European Union, Japan and the United States has to be revised in drinking water the arsenic standard is 10 g/L, the < > new standard for drinking water quality in China (July 1, 2006) will also be the standard for arsenic decreased from 50 g/L to 10 g/L.
The international agency for research on cancer (IARC) and the US Centers for Disease Control (CDC) has identified arsenic is the first class of carcinogens. Previous studies have shown that arsenic can cause skin cancer and lung cancer, and liver cancer, bladder cancer, kidney cancer, is also closely related to the incidence of colorectal cancer. In addition, studies have found that sodium arsenite (display NaAsO_2) with genetic toxicity, but the research on the mechanism of genotoxicity is not much. The results show that NaAsO_2 can lead to reactive oxygen species (ROS) generation cells increased and 8- hydroxydeoxyguanosine (8-OHdG) expression.
This study through the observation of different concentrations of NaAsO_2 on rat islet beta cell line INS-1 ROS and glutathione (GSH) level, influence the expression of 8-OHdG and DNA damage, arsenic caused injury of DNA cells from oxidative stress, to provide the basic data for the oxidative damage caused by arsenic and further research on the.
Methods: INS-1 cells were used as experimental objects, by single cell gel electrophoresis (SCGE) assay for detection of cell DNA damage and genetic toxicity evaluation NaAsO_2: genetic toxicity in order to explore the mechanism of arsenic, 2 ', 7' - two hydrogen chloride two fluorescein (DCFH-DA) method and two benzene aldehyde (OPT) in ROS cells and GSH levels were measured, the expression level was detected by immunohistochemistry in cell 8-OHdG. The experimental results with SPSS11.5 statistics.
Results: NaAsO_2 (2-16 mol/L) in INS-1 cells after 12h induced DNA strand breaks, cells form a comet like tail, tail and decreased the number of cells in a dose-dependent manner.NaAsO_2 (2-16 mol/L) in INS-1 cells after 12h may induce the expression of 8-OHdG increased; NaAsO_2 (2-16 mol/L) respectively. The role of 6H and 12h in INS-1 cells, may induce the expression of intracellular ROS levels increased significantly, in a dose-dependent manner, especially after 12h cells, the expression level of ROS was most significant; 8 mol/L and 16 mol/L NaAsO_2 in INS-1 cells after 24h, the expression of the intracellular ROS level increased (NaAsO_2; 2-16 mol/L) respectively in INS-1 cells after 48h, the expression level of.NaAsO_2 had no obvious change in ROS cells (2-16 mol/L) in INS-1 cells of 6,12,24 and 48h, may induce the expression of intracellular GSH levels increased significantly, in a dose-dependent manner.
Conclusion: NaAsO_2 induced DNA damage in INS-1 cells through oxidative stress mechanism.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R114

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