本文選題：砷化物　切入點：大鼠胰島β細胞　出處：《大連醫(yī)科大學》2013年碩士論文　論文類型：學位論文

【摘要】：目的：砷是普遍存在于自然界中毒性很強的類金屬元素，人類主要從飲水、食物和土壤中攝取砷，慢性砷中毒已成為全球面臨的重大公共衛(wèi)生問題。近年有流行病學研究顯示砷與糖尿病之間存在關聯(lián)，研究顯示砷導致糖尿病的主要機制可能是破壞胰島β細胞功能導致細胞損傷或凋亡。砷化物的化學形式分為無機砷和有機胂，目前研究表明，，砷化物的毒性高度依賴于它的化學形式，不同形態(tài)的砷在細胞內代謝途徑不同，具有不同的毒理學性質。盡管砷暴露作為糖尿病的危險因素已被認可，但迄今為止其生物效應機制仍未完全研究清楚，且實驗室數(shù)據(jù)多為研究三價無機砷與糖尿病之間的關聯(lián)。為了進一步的探索砷化物和糖尿病之間的關系和可能機制，本研究試圖觀察不同砷化物對體外培養(yǎng)大鼠胰島β細胞株（INS-1）的毒性作用，為砷導致糖尿病的發(fā)病機制研究提供線索。 方法：本實驗選定大鼠胰島β細胞株（INS-1），通過MTT法檢測砷化物對胰島細胞的毒性作用。用annexin V-FITC/PI流式細胞儀檢測法和Hoechst33258細胞染色法檢測不同砷化物致細胞凋亡情況。用2’,7’-二氫二氯熒光素（DCFH）檢測細胞內活性氧(ROS)的含量，用Western blot免疫印跡蛋白法檢測細胞內P53的蛋白含量變化。實驗結果用SPSS13.0統(tǒng)計軟件包進行統(tǒng)計分析。 結果：高劑量暴露五價無機砷、五價二甲基胂（分別50μmol/L、100μmol/L）能夠降低INS-1的細胞生存率，且iAs~(5-)的細胞毒性強于DMA~(5+)；iAs~(5+)（50-200μmol/L）和DMA~(5+)（100-400μmol/L）作用于INS-1細胞48h后，最高劑量暴露組均有凋亡細胞明顯的增多現(xiàn)象，這一實驗結果提示高濃度的iAs~(5+)和DMA~(5+)暴露使細胞發(fā)生凋亡；高劑量iAs~(5+)、DMA~(5+)暴露24h均可以誘發(fā)INS-1細胞內ROS水平的顯著升高，并且藥物的劑量存在依賴關系，提示在iAs~(5+)、DMA~(5+)作用于細胞的初期，細胞內發(fā)生了氧化應激反應；檢測發(fā)現(xiàn)經iAs~(5+)染毒的INS-1細胞核內p53蛋白表達增多，而經DMA~(5+)染毒的INS-1細胞核內p53蛋白表達無明顯變化。 結論：不同砷化物（砷酸氫二鈉、二甲基胂酸鈉）對大鼠胰島β細胞的毒性作用及機制不同。
[Abstract]:Objective: arsenic is a highly toxic metalloid element in nature. People mainly take arsenic from drinking water, food and soil. Chronic arsenism has become a major public health problem worldwide. Epidemiological studies in recent years have shown a link between arsenic and diabetes. Studies have shown that the main mechanism of arsenic induced diabetes may be the destruction of islet 尾 cell function leading to cell damage or apoptosis. The chemical forms of arsenide are inorganic arsenic and organic arsine. The toxicity of arsenide is highly dependent on its chemical form. Different forms of arsenic have different metabolic pathways in cells and have different toxicological properties, although arsenic exposure has been recognized as a risk factor for diabetes. However, the mechanism of its biological effect has not been fully studied, and most of the laboratory data are related to the relationship between trivalent inorganic arsenic and diabetes mellitus, in order to further explore the relationship and possible mechanism between arsenide and diabetes, This study attempts to observe the toxic effects of different arsenides on rat islet 尾 cell line INS-1 in vitro, which provides clues for the study of the pathogenesis of arsenic induced diabetes. Methods: rat islet 尾 cell line INS-1 was selected to detect the toxicity of arsenide to islet cells by MTT method. Apoptosis induced by different arsenic compounds was detected by annexin V-FITC / Pi flow cytometry and Hoechst33258 cell staining. The content of reactive oxygen species (Ros) in the cells was determined by DCFH- DCFH- Dichlorofluorescein dihydrodichlorofluorescein (DCFH). The changes of p53 protein in cells were detected by Western blot Western blot method. The results were analyzed by SPSS13.0 software package. Results: high dose exposure to pentavalent inorganic arsenic, pentavalent dimethyl arsine (50 渭 mol / L, 100 渭 mol / L, respectively) decreased the survival rate of INS-1 cells, and the cytotoxicity of iAsN 5 was stronger than that of DMA~(5 (50 ~ 200 渭 mol / L) and DMA~(5 (100 ~ 400 渭 mol / L) for 48 h. The results showed that high concentration of iAs~(5) and DMA~(5) could induce apoptosis of INS-1 cells, and high dose of iAs~(5 could induce the increase of ROS level in INS-1 cells after 24 hours of exposure. Moreover, the dose-dependence of the drug indicated that the oxidative stress reaction occurred in the cells at the early stage of the action of iAs~(5, and the expression of p53 protein in the nucleus of INS-1 exposed to iAs~(5) was increased. However, the expression of p53 protein in the nucleus of INS-1 exposed to DMA~(5 had no significant change. Conclusion: the toxicity and mechanism of different arsenides (disodium arsenate, dimethylarsonic acid) on islet 尾 cells of rats are different.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R114

【參考文獻】

相關期刊論文 前10條

1 楊從容;王雅棣;;活性氧、錳超氧化物歧化酶和p53的相互作用及對腫瘤的影響[J];癌變·畸變·突變;2011年03期

2 陸景坤;陳朝軍;翟小涵;李小東;;不同砷化合物對人皮膚成纖維細胞系的毒性研究[J];中國地方病防治雜志;2011年05期

3 洪峰;無機砷的腎臟損傷作用研究進展[J];國外醫(yī)學(衛(wèi)生學分冊);2002年01期

4 陸景坤;陳朝軍;劉小雷;李小東;翟小涵;;不同砷化合物對人胚腎細胞系HEK-293細胞的毒性研究[J];環(huán)境與健康雜志;2011年01期

5 陸景坤;田艷;俞騰飛;陳朝軍;王一博;尹若熙;;砷化合物致人表皮癌A431細胞的毒性及氧化應激和砷代謝研究[J];環(huán)境與健康雜志;2012年09期

6 彭珊茁;蔡原;;砷的生物學指標研究進展[J];中國工業(yè)醫(yī)學雜志;2006年03期

7 張冰蔭;慕曉玲;;砷引發(fā)糖尿病的研究進展[J];現(xiàn)代生物醫(yī)學進展;2009年17期

8 徐躍飛;任鳳;趙寶昌;;Adinbitor對人胃癌MGC803細胞增殖及凋亡的作用[J];實用癌癥雜志;2009年03期

9 李冰;陸春偉;孫貴范;;不同砷化合物對血管內皮細胞毒性的研究[J];衛(wèi)生研究;2006年06期

10 孫志;馬麗;邱玉芹;韓海容;;2型糖尿病發(fā)病機制及胰島β細胞功能障礙的研究進展[J];醫(yī)學綜述;2008年09期

本文編號：1585639