發(fā)布時間：2018-03-28 18:55

  本文選題：亞砷酸鈉　切入點：大鼠胰島細胞　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、日本和美國等先后將飲用水中砷的標準修訂為10μg/L，我國新的《生活飲用水衛(wèi)生標準》（2006年7月1日實施）也將砷含量標準由50μg/L降低到10μg/L。 國際癌癥研究機構(gòu)（IARC）和美國疾病控制中心（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)遺傳毒性機制的研究不多。研究表明，NaAsO_2可導(dǎo)致細胞中活性氧類（ROS）生成增多和8-羥基脫氧鳥苷（8-OHdG）表達升高。 本研究通過觀察不同濃度NaAsO_2對大鼠胰島β細胞株INS-1內(nèi)ROS和谷胱甘肽（GSH）水平、8-OHdG表達和DNA損傷的影響的影響，從氧化應(yīng)激角度探討砷所導(dǎo)致細胞DNA損傷的可能機制，為進一步的深入研究砷所引起的機體氧化損傷提供基礎(chǔ)資料。 方法：以INS-1細胞作為實驗對象，通過單細胞凝膠電泳（SCGE）試驗檢測細胞DNA損傷情況，，評價NaAsO_2的遺傳毒性：為探討砷的遺傳毒性機制，以2’,7’-二氫二氯熒光素（DCFH-DA）法和苯二醛（OPT）分別測定細胞內(nèi)ROS以及GSH水平，以免疫組化方法檢測細胞內(nèi)8-OHdG的表達水平。實驗結(jié)果用SPSS11.5統(tǒng)計。 結(jié)果：NaAsO_2（2-16μmol/L）作用于INS-1細胞12h后，可引起DNA鏈斷裂，細胞形成彗星樣拖尾，拖尾細胞數(shù)明顯增多，且呈劑量依賴關(guān)系。NaAsO_2（2-16μmol/L）作用于INS-1細胞12h后，可致8-OHdG的表達升高；NaAsO_2（2-16μmol/L）分別作用于INS-1細胞6h和12h后，可致細胞內(nèi)ROS表達水平的顯著增加，且呈劑量依賴關(guān)系，尤其是作用12h后，細胞內(nèi)ROS表達水平最為顯著；8μmol/L和16μmol/L NaAsO_2作用于INS-1細胞24h后，細胞內(nèi)ROS表達水平增加；NaAsO_2（2-16μmol/L）分別作用于INS-1細胞48h后，細胞內(nèi)ROS表達水平未見明顯改變。NaAsO_2（2-16μmol/L）作用于INS-1細胞6，12，24和48h后，可致細胞內(nèi)GSH表達水平的顯著增加，且呈劑量依賴關(guān)系。 結(jié)論：NaAsO_2通過氧化應(yīng)激機制引起INS-1細胞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)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R114

【參考文獻】

相關(guān)期刊論文 前9條

1 李玲;吳君;蔣玲;歐兵;張韻;李誠秀;程明亮;楊勤;;不同價態(tài)飲水砷暴露對小鼠肝臟的損傷作用[J];第二軍醫(yī)大學(xué)學(xué)報;2008年05期

2 王惠惠;徐苑苑;李冰;李昕;薛鵬;賀淼;王毅;孫貴范;;亞砷酸鈉致Chang liver細胞氧化應(yīng)激作用的研究[J];環(huán)境與健康雜志;2008年07期

3 宋歌;崔熠;夏紅飛;馬旭;;亞砷酸鈉致早期雞胚胚胎發(fā)育毒性與基因甲基化的關(guān)系[J];中國計劃生育學(xué)雜志;2011年05期

4 凈錦飛;徐焰;駱文靜;劉明朝;沈?qū)W峰;陳耀明;張建彬;趙芳;鄭剛;陳景元;;亞慢性砷暴露對大鼠腎臟功能和形態(tài)的影響[J];實用預(yù)防醫(yī)學(xué);2011年02期

5 康家琦;砷對健康危害的研究進展[J];衛(wèi)生研究;2004年03期

6 楊運旗,孫蘭英,張碧霞,張愛華;慢性燃煤污染型砷中毒患者腎功能改變[J];微量元素與健康研究;2000年02期

7 姚芹;吳順華;湯紅英;馬艷;吳軍;張杰;;亞砷酸鈉亞慢性染毒對小鼠肝臟腎臟的損傷作用[J];新疆醫(yī)科大學(xué)學(xué)報;2010年12期

8 李遠慧;朱筑霞;李娜;吳澤江;;慢性砷中毒對腎小球細胞凋亡形態(tài)學(xué)影響[J];中國公共衛(wèi)生;2008年09期

9 洪峰,金泰^

本文編號：1677608