本文選題：氟 + 腫瘤�。� 參考：《廣西醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的探索氟的遺傳毒性及維生素C(VC)、維生素E(VE)、維生素C聯(lián)合維生素E (VC+VE)對染氟淋巴細(xì)胞損傷的干預(yù)作用效果。 方法1.職業(yè)環(huán)境氟污染調(diào)查：以鋁電解廢氣凈化車間為污染源中心,在不同半徑距離的工作位點設(shè)采樣點檢測空氣中的氟,用氟離子選擇電極法測定其含量。2.職業(yè)性氟接觸與腫瘤發(fā)生的關(guān)系調(diào)查：通過問卷調(diào)查和醫(yī)院病歷收集該廠職工從1995～2009年底腫瘤發(fā)生情況,進(jìn)行統(tǒng)計分析。3.氟的遺傳損傷動物實驗研究：(1)檢測經(jīng)低、中、高濃度(2mg·kg-1·d-18mg·kg-1·d-1、32mg·kg-1·d-1)氟化鈉染毒14d后小鼠骨髓細(xì)胞微核率,(2)用衰減全反射傅立葉變換紅外光譜技術(shù)(ATR-FTIR)在包括RNA/DNA標(biāo)識波長(1020/1121cm-1)在內(nèi)的波段進(jìn)行掃描,檢測小鼠肝細(xì)胞核化學(xué)結(jié)構(gòu)的變化。4.氟的遺傳損傷及VC、VE干預(yù)作用的體外細(xì)胞培養(yǎng)實驗研究：提取正常人外周血淋巴細(xì)胞,培養(yǎng)48h后分組及處理：(1)用0.00、0.01、0.04、0.16、0.64mg/ml NaF(對照組、F1、F2、F3、F4)分別作用2h、4h、8h、16h后, MTT法測定淋巴細(xì)胞存活率。(2)淋巴細(xì)胞分為對照組、F1、F2、F3、F4,低、中、高VC劑量干預(yù)(VC1+F4、VC2+F4、 VC3+F4)組,低、中、高VE劑量干預(yù)(VE1+F4、VE2+F4、VE3+F4)組,低、中、高VC+VE劑量干預(yù)(VC1+VE1+F4、VC2+VE2+F4、 VC3+VE3+F4)組。對照組給予培養(yǎng)液28h, F1、F2、F3、F4組給予培養(yǎng)液24h后分別暴露于含0.01、0.04.0.16、0.64mg/ml NaF培養(yǎng)液4h, VC干預(yù)組分別于含4.16、64μmol/LVC培養(yǎng)液作用24h后暴露于0.64mg/mlNaF培養(yǎng)液4h, VE干預(yù)組分別于含2、10、50μmol/L VE培養(yǎng)液作用24h后暴露于0.64mg/ml NaF培養(yǎng)液4h, VC+VE干預(yù)組分別于含4+2、16+10、64+50μmol/L VC+VE培養(yǎng)液作用24h后暴露于0.64mg/ml NaF培養(yǎng)液4h,離心收集細(xì)胞,分別用MTT法測淋巴細(xì)胞存活率,Annexin V-FITC細(xì)胞凋亡試劑盒檢測細(xì)胞凋亡率,單細(xì)胞凝膠電泳技術(shù)(SCGE)測定DNA的損傷,Elisa實驗法檢測細(xì)胞端粒酶含量。 結(jié)果 1.職業(yè)環(huán)境氟污染調(diào)查結(jié)果：以氟污染源鋁電解廢氣凈化車間為中心,在不同半徑距離的工作位點空氣中的氟濃度依次降低,呈現(xiàn)氟污染擴(kuò)散濃度與距離的負(fù)向趨勢,但均沒有超過國家衛(wèi)生標(biāo)準(zhǔn)(0.50mg/m3)。 2.職業(yè)性氟接觸與腫瘤發(fā)生的關(guān)系調(diào)查：廠區(qū)工人腫瘤粗發(fā)病率為117.95/10萬(標(biāo)化率為58.81/10萬),標(biāo)化發(fā)病率女性比男性高(男：女=1：2.64),腫瘤發(fā)病年齡高峰為40-49歲,前2位腫瘤男性為肝癌和肺癌,女性為乳腺癌和肺癌。與該地區(qū)非暴露人群比較,該鋁廠工人的氟暴露可能使女性腫瘤發(fā)病率升高,為該市的2.14倍,腫瘤發(fā)病年齡提前,腫瘤構(gòu)成基本相同。 3.氟的遺傳損傷動物實驗結(jié)果： 3.1動物實驗小鼠骨髓細(xì)胞微核率隨染氟濃度的增高而升高。 3.2肝細(xì)胞核ATR光譜圖呈1650cm-1、1550cm-1、1380cm-1、1260cm-1、1225cm-1、1155cm-1、1080cm-1、1030cm-1、970cm-1、930cm-1峰降低；光譜數(shù)據(jù)用統(tǒng)計軟件進(jìn)行主成分分析和3-D圖形處理后,可以觀察到更細(xì)微的化學(xué)結(jié)構(gòu)變化,與對照組比較,氟中、高濃度組的差異以第1主成分因子為主,而氟低濃度組的差異以第2主成分因子為主,顯示不同氟濃度對小鼠肝細(xì)胞核的化學(xué)結(jié)構(gòu)影響存在明顯差異。 4.氟的遺傳損傷及VC、VE干預(yù)作用的體外細(xì)胞培養(yǎng)實驗研究： 4.1細(xì)胞存活率隨染氟濃度與時間的增加而下降。與對照組相比,F4組細(xì)胞存活率顯著下降(P0.05)。16h各處理組細(xì)胞存活率均低于2h處理組(P0.05)。 4.2淋巴細(xì)胞染氟4h后,(1)總凋亡率隨染氟濃度增加而升高,與對照組相比,總凋亡率分別增加了2.47%、9.80%、13.52%、25.74%,除F1濃度組外,其余各組增加均有統(tǒng)計學(xué)意義(P0.05)；(2)與對照組相比,隨著NaF染毒濃度的升高,拖尾細(xì)胞數(shù)增加,彗星頭部變小,亮度增加,彗尾變長變圓,熒光強(qiáng)度增強(qiáng),其尾部DNA百分率、尾長、Olive尾矩升高呈劑量依賴性(P0.01)；(3)端粒酶含量隨染氟濃度增加而升高。與對照組相比,F3、F4濃度組端粒酶含量升高有統(tǒng)計學(xué)意義(P0.05),且F4濃度組端粒酶含量明顯高于其余各組(P0.01)。 4.3經(jīng)預(yù)防性干預(yù)后的各組與F4組比較,(1) VC、VE、VC+VE各干預(yù)組淋巴細(xì)胞存活率均有升高的趨勢。VC各組淋巴細(xì)胞存活率隨著濃度的增加而升高,其中VC3+F4組細(xì)胞存活率升高有統(tǒng)計學(xué)意義(P0.05),聯(lián)合作用VC2+VE2+F4組細(xì)胞存率升高最顯著(P0.05)；(2)VC、VE、VC+VE各干預(yù)組淋巴細(xì)胞總凋亡率均有降低的趨勢。VC、VE各干預(yù)組淋巴細(xì)胞總凋亡率隨著濃度的增加而降低,VC2+F4、VC3+F4、 VE3+F4組淋巴細(xì)胞總凋亡率降低有統(tǒng)計學(xué)意義(P0.05)。單純VC、VE各總凋亡率低于VC+VE各干預(yù)組,但無統(tǒng)計學(xué)差異(P0.05)；(3)VC、VE、VC+VE各干預(yù)組彗星頭部變大,彗尾變短,尾部DNA百分率、尾長、Olive尾矩均顯著降低(P0.01),VE1組的Olive尾矩最短。VC+VE干預(yù)組均高于VC、VE各組尾部DNA百分率、尾長和Olive尾矩(P0.05)；(4)VC、VE、VC+VE各干預(yù)組淋巴細(xì)胞端粒酶含量均有降低。VC、VE各組淋巴細(xì)胞端粒酶含量隨著濃度的增加而降低,VC3+F4、VE3+F4濃度組淋巴細(xì)胞端粒酶含量降低有統(tǒng)計學(xué)意義(P0.05)。 VC+VE各干預(yù)組以VC2+VE2+F4、VC3+VE3+F4組淋巴細(xì)胞端粒酶含量降低有統(tǒng)計學(xué)意義(P0.05),且VC2+VE2+F4濃度組降低更顯著(P0.01)。 結(jié)論 在本研究條件下, 1.職業(yè)性氟接觸組女性腫瘤發(fā)病率升高,腫瘤發(fā)病年齡提前,氟可能是一種潛在的致癌物。 2.氟可致實驗動物小鼠染色體畸變、RNA/DNA結(jié)構(gòu)的改變和體外培養(yǎng)人淋巴細(xì)胞細(xì)胞凋亡、DNA損傷、端粒酶含量升高,具有明顯的遺傳損傷效應(yīng),可能與腫瘤發(fā)生有關(guān)。 3.在一定劑量范圍內(nèi),VC、VE對氟致淋巴細(xì)胞損傷有明顯的干預(yù)作用。 4. ATR-FTIR可敏感地檢測環(huán)境污染物引起的RNA、DNA.蛋白質(zhì)等物質(zhì)的改變,可作為早期遺傳損傷監(jiān)測的方法之一。
[Abstract]:Objective to explore the genetic toxicity of fluoride and vitamin C (VC), vitamin E (VE), vitamin C and vitamin E (VC+VE) on the intervention effect of fluoride on lymphocytes.
Investigation of 1. occupation of environmental fluoride pollution method: aluminum electrolytic gas purification workshop for pollution source, sampling points were set in the air fluoride working positions at different radii, investigate the relationship to determine the content of.2. occupation exposed to fluorine and tumor by fluoride ion selective electrode method: through the questionnaire survey and the hospital medical records factory workers from the end of 1995~2009 the tumor incidence of genetic damage in animal experimental study of statistical analysis: (1).3. fluoride detection by low, high concentration (2mg - kg-1 - d-18mg - kg-1 - d-1,32mg - kg-1 - D-1) mice bone marrow cell micronucleus rate of sodium fluoride exposure after 14d (2) with attenuation total reflection Fu Liye transform infrared spectroscopy (ATR-FTIR) in the RNA/DNA logo (1020/1121cm-1), wavelength band scanning, genetic damage and VC fluoride.4. to detect changes in the chemical structure of mouse liver nuclei, VE intervention Experimental study of cultured cells in vitro: peripheral blood lymphocytes from normal people, grouping and treatment after 48h culture: (1) 0.00,0.01,0.04,0.16,0.64mg/ml NaF (control group, F1, F2, F3, F4) respectively. 2h, 4h, 8h, 16h, MTT lymphocyte survival rate determination method. (2) lymphocytes into the control group, F1, F2, F3, F4, low, high dose of VC (VC1+F4, VC2+F4, VC3+F4 intervention group), low, high dose of VE (VE1+F4, VE2+F4, VE3+F4 intervention group), low, high dose of VC+VE (VC1+VE1+F4, VC2+VE2+F4, VC3+ intervention VE3+F4) control group. 28h group received medium, F1, F2, F3, F4 group received medium 24h respectively after exposure to NaF medium containing 0.01,0.04.0.16,0.64mg/ml 4H and VC 4.16,64 respectively in the intervention group containing mol/LVC medium 24h after exposure to 0.64mg/mlNaF medium 4h, VE group respectively with 2,10,50 mol/L VE medium 24h after exposure to 0.64mg/ml NaF culture Liquid 4h, VC+VE group respectively with 4+2,16+10,64+50 mol/L VC+VE cultured 24h after exposure to 0.64mg/ml NaF medium 4h, cells were collected, respectively. MTT method was used to test lymphocyte survival, apoptotic cells were detected by Annexin V-FITC apoptosis kit, single cell gel electrophoresis (SCGE) determination of DNA damage. Determination of telomerase Elisa assay.
Result
1. occupation of environmental fluoride pollution survey results: the fluorine pollution source of aluminum electrolysis gas purification plant as the center, the fluoride concentration in different radial distance in the air in order to reduce the work site, the negative trend of fluorine pollution diffusion concentration and distance, but did not exceed the national hygienic standard (0.50mg/m3).
Investigation on the relationship between the 2. occupation exposed to fluorine and cancer: the workers of crude cancer incidence rate of 117.95/10 million (the standardized rate was 58.81/10 million), the standardized incidence rate was higher in females than in males (male: female =1:2.64), the peak age of onset of tumors was 40-49 years ago, 2 tumors were liver cancer and lung cancer in men, women for breast cancer and lung cancer. And the area of non exposed population, the factory workers exposed to fluoride may make women increased tumor incidence, is 2.14 times the city's tumor early age of onset, tumor structure is basically the same.
3. fluorine genetic damage in animal experiments:
3.1 animal experimental mouse bone marrow cell micronucleus rate increased with the increase of fluoride concentration increased.
3.2 liver nuclear ATR spectra was decreased 1650cm-11550cm-11380cm-11260cm-11225cm-11155cm-11080cm-11030cm-1970cm-1930cm-1 peak spectral data; principal component analysis and 3-D graphics processing with statistical software, can be observed that the chemical structure of the more subtle change, compared with control group, fluoride, the difference of the high concentration group with first principal component factors, and the difference of fluoride and low concentration group based on second principal component factors, the effects of different fluoride concentration and chemical structure of the mouse liver cell nuclei were significantly different.
The genetic damage and VC 4. fluorine, experimental study of effects of VE on the in vitro cell culture:
4.1 cell survival rate increased with the increase of fluoride concentration and time decreased. Compared with the control group, F4 group significantly decreased cell viability in each treatment group (P0.05).16h cell survival rate were lower than 2H treatment group (P0.05).
4.2 lymphocyte fluoride 4h, (1) the total apoptosis rate with fluoride concentration increased, compared with the control group, the apoptosis rate were increased by 2.47%, 9.80%, 13.52%, 25.74%, in addition to the concentration of F1 group, the other groups were statistically significant increased (P0.05); (2) compared with the control group. With the increase of NaF concentrations, the trailing cell number increased, the comet head becomes small, increased brightness, long tail round, fluorescence intensity, the percentage of tail DNA, tail length, Olive tail moment increased in a dose dependent manner (P0.01); (3) telomerase content increased with increasing concentration. Compared with fluoride with the control group, F3 group, F4 concentration increased telomerase content was statistically significant (P0.05), and the concentration of F4 group was significantly higher than that of other groups of telomerase (P0.01).
The 4.3 group compared with the F4 group intervention of prevention, (1) VC, VE, VC+VE lymphocyte survival rate in each intervention group increased.VC lymphocyte survival rate were increased with increasing concentration, the survival rate of cells in VC3+F4 group increased significantly (P0.05), the combined effect of VC2+VE2+F4 group cell survival rate the most significant increase (P0.05); (2) VC, VE.VC VC+VE, the trend in each intervention group were decreased the apoptosis rate of lymphocytes, VE lymphocytes apoptosis rate in each intervention group with the concentration increased, VC2+F4, VC3+F4, VE3+F4 group had a significant decrease in apoptosis rate of lymphocytes (P0.05). Only VC. VE the total apoptosis rate lower than the intervention group VC+VE, but no significant difference (P0.05); (3) VC, VE, VC+VE of the intervention group head of the comet tail becomes larger and shorter, tail DNA, tail length, Olive tail moment were significantly decreased (P0.01), VE1 group, Olive tail moment most Short.VC+VE intervention group were higher than VC, VE groups of tail DNA, tail length and Olive tail moment (P0.05); (4) VC, VE, VC+VE of the intervention group had decreased.VC lymphocyte telomerase, VE lymphocytes were telomerase content decreased with the increase of the concentration of VC3+F4, VE3+ concentration of F4 group decreased the content of lymphocyte telomerase there was statistical significance (P0.05). VC+VE VC2+VE2+F4 in the intervention group, VC3+VE3+F4 group decreased the content of lymphocyte telomerase was statistically significant (P0.05), and the concentration of VC2+VE2+F4 group decreased more significantly (P0.01).
conclusion
Under the condition of this study,
The group of women increased tumor incidence of 1. occupation of fluoride exposure, tumor early age of onset, fluoride is a potential carcinogen.
2. fluorine can experimental animal mouse chromosome aberration, lymphocyte apoptosis, and in vitro culture change the structure of RNA/DNA DNA injury, the telomerase content increased, with significant genetic damage, may be related to tumorigenesis.
3. in a certain dose range, VC, VE has a significant intervention effect on lymphocyte damage induced by fluoride.
4. ATR-FTIR can sensitively detect the environmental pollution caused by RNA, the changes of substance DNA. protein, can be used as one of the methods for monitoring genetic damage in early stage.

【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 邵紅,劉開泰,姚華,張躍新,劉文亞,冀芳,張婕;氟中毒患者外周血淋巴細(xì)胞基因表達(dá)水平的研究[J];地方病通報;2005年02期

2 劉早玲;賈漢;巴杭;多力坤;;氟對大鼠肝腎磷脂脂肪酸組成的影響及維生素C拮抗作用的研究[J];中國地方病防治雜志;2006年02期

3 翟鵬勇;曾錦榮;譚寧;王績英;黃嵐珍;佘巍巍;;維生素C對A549細(xì)胞增殖、凋亡及Caspase-3、Survivin表達(dá)的影響[J];中國肺癌雜志;2010年02期

4 黃意府;王清海;黃鮮華;;低濃度氟作業(yè)工人的外周血淋巴細(xì)胞染色體畸變分析[J];廣西預(yù)防醫(yī)學(xué);2006年01期

5 楊莉萍;;維生素E和維生素C對HL-60細(xì)胞NO生成的影響[J];海南醫(yī)學(xué);2007年05期

6 王寧;王俐;張慧君;孫靜秋;鄭衛(wèi)東;肖萍;;氟化鈉對大鼠軟骨細(xì)胞活性的影響及DNA損傷作用[J];環(huán)境與健康雜志;2010年10期

7 葛穎華;鐘曉明;;維生素C和維生素E抗氧化機(jī)制及其應(yīng)用的研究進(jìn)展[J];吉林醫(yī)學(xué);2007年05期

8 榮熙敏;;端粒酶與細(xì)胞衰老和癌變的研究[J];內(nèi)江科技;2011年01期

9 高希寶,李文生,史麗,許之,張寧,朱振平;氟化物對 DNA 損傷的機(jī)理探討[J];山東醫(yī)藥;1997年05期

10 謝萍;自由基與細(xì)胞凋亡[J];生物學(xué)教學(xué);2004年01期

相關(guān)碩士學(xué)位論文 前1條

1 趙慧東;維生素E和鈣劑對大鼠氟牙癥發(fā)生的拮抗作用研究[D];山西醫(yī)科大學(xué);2011年

，

本文編號：1745840