本文選題：氟化鈉　切入點：人體骨髓瘤細胞　出處：《中央民族大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：氟是人體必需的微量元素之一,對人體健康具有兩面性。適量的氟攝入,有利于牙齒和骨骼生長,維持體內(nèi)鈣磷平衡,同時能有效預(yù)防齲齒；但長期過量氟暴露時,氟逐漸在體液和組織中蓄積,對機體產(chǎn)生毒性作用,引起嚴重的健康問題,典型臨床癥狀為氟骨癥和氟斑牙。目前,地氟病仍是我國許多地區(qū)最嚴重的公共衛(wèi)生問題之一,除上海市和海南省外,其余各省、區(qū)幾乎都有不同程度的發(fā)生,尤其在廣大農(nóng)村地區(qū)和邊遠山區(qū)。長期以來,人們對于氟暴露產(chǎn)生的骨相損傷進行了大量研究,逐漸以氟斑牙、氟骨癥作為氟暴露效應(yīng)終端,但后來發(fā)現(xiàn)具有一定局限性。氟斑牙和氟骨癥均屬于分類、跳躍型變量,不具備連續(xù)性。以此類指標和暴露濃度進行擬合時,由于評估指標的鈍化和間斷性,將直接影響推導(dǎo)結(jié)果的準確性。因此,為了探索氟暴露劑量-效應(yīng)關(guān)系,尋求早期效應(yīng)標志,本研究開展了細胞和動物氟化鈉染毒實驗,具體內(nèi)容如下：(1)以人體骨髓瘤細胞RPMI8226為研究對象,用氟化鈉進行染毒,建立細胞氟暴露模型。染毒濃度分別為0、10、20、40、80、160、320μM,染毒時間為48 h。染毒結(jié)束后,提取細胞RNA,運用RT-QPCR技術(shù)測定基因表達變化(上調(diào)或下調(diào))。結(jié)果顯示,與轉(zhuǎn)錄有關(guān)、與蛋白合成降解有關(guān)以及與細胞增殖、凋亡和周期有關(guān)的三類基因的表達受到氟暴露不同程度影響。其中,與轉(zhuǎn)錄有關(guān)的基因ANKRD1、KLF2、SBN02和ZNF649,與空白對照組相比,在高濃度染毒組(320 μM), mRNA相對表達量顯著上升(P0.01)。與蛋白合成降解有關(guān)的基因FANCM的mRNA相對表達量在80、320μM染毒組明顯上升。而與細胞增殖、凋亡和周期有關(guān)的基因PDGFA. RNF152、CDK10和CETN2,當染毒濃度大于160μM時,mRNA相對表達量相對于空白對照組顯著上升(P0.01)。(2)以SD大鼠為研究對象,通過灌胃方式進行氟化鈉染毒,建立動物慢性氟中毒模型。染毒劑量分別為0、2、4、8、16、32 mg/kg. b. w,染毒時間為3個月。染毒期間,隔天對大鼠進行稱重,并觀察氟斑牙發(fā)生情況。染毒結(jié)束后,測定動物常規(guī)指標、生化指標以及特異性蛋白指標變化情況。實驗結(jié)果顯示,氟化鈉暴露對大鼠體重有一定影響。與空白對照組相比,隨著氟化鈉染毒劑量的增加,大鼠體重增長率逐漸下降。對于大鼠臟器系數(shù),隨著氟化鈉染毒劑量的增大,心臟系數(shù)和胸腺系數(shù)逐漸減小,而肝臟系數(shù)逐漸增大。當染毒劑量為16、32mg/kg. b. w時,與空白對照組相比,大鼠肝臟系數(shù)顯著增大(P0.05),表明兩個高氟劑量組大鼠肝臟受到一定程度損傷。再有,肝功能生化指標檢測顯示,AST活力隨染毒濃度的增加有降低趨勢,當染毒劑量大于4 mg/kg. b. w,大鼠AST活力明顯降低,差異具有統(tǒng)計學(xué)意義。最后,大鼠血液中特異性蛋白活性測定結(jié)果顯示,隨著氟暴露濃度的增大,蛋白GLRa2和RYR1含量沒有顯著變化；而蛋白ATF4的含量隨著暴露濃度的增大,出現(xiàn)顯著下降的趨勢,并且蛋白含量和暴露濃度之間呈現(xiàn)顯著負相關(guān)(R=-0.586,P0.01)。因此,可考慮將血液中ATF4含量作為流行病學(xué)效應(yīng)指標,用于預(yù)測早期高氟暴露。本論文在成功建立細胞和動物氟暴露實驗?zāi)Ｐ偷幕A(chǔ)上,測定了生物效應(yīng)指標的變化,研究探討了氟暴露劑量-效應(yīng)關(guān)系,為氟暴露早期健康效應(yīng)研究提供了建議,同時為地方性氟病的預(yù)防和治療提供了科學(xué)依據(jù)。
[Abstract]:Fluorine is one of essential trace elements in human body, has two sides to human health. The amount of fluoride intake, is conducive to the growth of teeth and bones, to maintain the balance of calcium and phosphor, and can effectively prevent dental caries; but the long-term exposure to excessive fluoride, fluoride gradually accumulate in body fluids and tissues, produce toxic effects on the body, causing health the problem is serious, the typical clinical symptoms of skeletal fluorosis and dental fluorosis. At present, one of the most serious public health problems in many areas of endemic fluorosis in China is still in the provinces of Shanghai and Hainan provinces, area almost all have occurred in different degree, especially in the vast rural areas and remote mountainous areas. For a long time, the fluoride exposure caused bone damage has been studied, and gradually to dental fluorosis, skeletal fluorosis as fluoride exposure effect of terminal, but later found that it has certain limitations. Dental fluorosis and skeletal fluorosis are classified, jump Step type variable does not have continuity. By fitting to such indicators and exposure concentration, because the passivation evaluation index and discontinuity, the accuracy will directly affect the results. Therefore, in order to explore the dose-response relationship between the fluoride exposure, seeking early effect markers, this study developed the cell and animal experiment of sodium fluoride exposure and the specific contents are as follows: (1) in human myeloma cell RPMI8226 as the research object, were treated with sodium fluoride, fluoride exposure to establish cell model. Exposure concentration was 0,10,20,40,80160320 M, exposure time of 48 h. exposure after the extraction of RNA cells, determination of gene expression using RT-QPCR Technology (up or down). The results showed that transcription, protein synthesis and degradation is related with cell proliferation, apoptosis and gene expression of three kinds of cycle related to the different influence of fluoride exposure. Among them, with relevant transcription The gene ANKRD1, KLF2, SBN02 and ZNF649, compared with the control group, the high concentration exposure group (320 M), the relative expression level of mRNA increased significantly (P0.01). FANCM gene related to protein synthesis and degradation of the relative expression of mRNA increased significantly in the 80320 M exposure group. The cell proliferation and, cycle and apoptosis related genes PDGFA., RNF152, CDK10 and CETN2, when the exposure concentration is greater than 160 M, the relative expression of mRNA compared with the control group increased significantly (P0.01). (2) with SD rats as the research object, were exposed by intragastric administration of sodium fluoride, establish the animal model of chronic fluorosis at doses of 0,2,4,8,16,32 mg/kg.. B. W, the exposure time was 3 months. During the exposure, the rats were weighed and observed the occurrence of dental fluorosis. After exposure, determination of conventional indicators of animal, biochemical indexes and specific protein index changes. Experimental results Results showed that sodium fluoride exposure has a certain effect on the body weight of rats. Compared with the control group, with the increase of sodium fluoride dose, rat body weight growth rate decreased gradually. The organ coefficient of rats, with the increase of sodium fluoride dose, cardiac index and thymus index was gradually decreased, and the liver coefficient increases gradually when the dose was 16,32mg/kg.. B. W, compared with the control group, the liver index of rats increased significantly (P0.05), two showed that the high fluoride dose group rat liver by a certain degree of damage. Furthermore, detection of biochemical indexes of liver function showed that AST activity decreased with exposure concentration decreased with the increase of exposure, when more than 4 mg/kg. B. dose of W, activity of AST rats decreased significantly, the difference was statistically significant. Finally, results show that the determination of specific protein activity in blood of rats exposed to fluoride, with the increasing concentration of egg white, GLRa2 and RYR1 were not There are significant changes; and the content of ATF4 protein increased with the exposure concentration, significantly decreased, and between protein content and exposure concentration showed a significant negative correlation (R=-0.586, P0.01). Therefore, the content of ATF4 in blood can be considered as epidemiological effect index for the prediction of early high fluoride exposure. This thesis established cell and the animal experiment model of fluoride in successful exposure, changes of biological effect index determination, study the dose effect relationship of fluoride exposure, provides suggestions on health effects of early exposure to fluoride, and provide scientific basis for prevention and treatment of endemic fluorosis.

【學(xué)位授予單位】：中央民族大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R599.1

【參考文獻】

相關(guān)期刊論文 前8條

1 劉丹;祁永紅;王克躍;周遠忠;石修權(quán);;燃煤型氟暴露地區(qū)人群尿氟水平及影響因素[J];環(huán)境與健康雜志;2014年03期

2 王平貴;王俊玲;剛亞棟;張本忠;;慢性氟中毒對大鼠腎功能的影響及其與氧化應(yīng)激的關(guān)系[J];毒理學(xué)雜志;2009年05期

3 楊永建;;氟對人體健康的影響[J];中國科技信息;2007年09期

4 于燕妮,曹序茂,肖開棋,劉萬興,董仲,茹景順;慢性氟中毒大鼠肝臟損傷的形態(tài)計量學(xué)研究[J];中國地方病學(xué)雜志;1990年01期

5 于燕妮,王守立,高勤,劉家騮,梁任又,金嗣昭,徐云華;應(yīng)用電子自旋共振技術(shù)檢測染氟大鼠骨肝腎組織中自由基含量[J];中國地方病學(xué)雜志;2001年05期

6 呂曉紅,呂曉麗,李廣生;慢性氟中毒鼠神經(jīng)元與細胞凋亡相關(guān)基因的研究[J];中國地方病學(xué)雜志;2003年02期

7 郭曉英,孫貴范,孫憲民,張穎,李昕,劉珊,孫鮮策,王毅;氧化應(yīng)激在亞慢性氟中毒大鼠肝臟損傷中的作用[J];中國公共衛(wèi)生;2003年05期

8 馬寶慧;潘桂蘭;陳曉東;呂軍;時靜華;;慢性氟中毒對大鼠血清氧化應(yīng)激反應(yīng)的影響[J];中國現(xiàn)代醫(yī)藥雜志;2012年09期

相關(guān)博士學(xué)位論文 前1條

1 向全永;環(huán)境中氟對居民健康影響的危險度評價[D];復(fù)旦大學(xué);2004年

，

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