本文選題：慢性氟中毒 + 大鼠��； 參考：《貴陽醫(yī)學(xué)院》2015年碩士論文

【摘要】：目的:觀察慢性氟中毒動物腦組織及體外培養(yǎng)細(xì)胞模型中主要炎癥因子腫瘤壞死因子a(tumor necrosis factora,TNFa)和白細(xì)胞介素-1b(interleukin-1b,IL-1b)的表達改變及細(xì)胞核因子-κB(nuclear factorκB,NF-κB)信號轉(zhuǎn)導(dǎo)通路中NF-κB、NF-κB抑制蛋白α(inhibitor of nuclear factorκBα,IκBα)的表達,探討其在慢性氟中毒炎性腦損傷機制中的作用。方法:建立慢性氟中毒動物模型:SD大鼠60只,雌雄各半,隨機分為正常對照組、染氟組(飲水氟含量50 mg/L,加入氟化鈉)2組;實驗期為10個月;氟中毒細(xì)胞模型:體外培養(yǎng)THP-1細(xì)胞,分為3組,即正常對照組、低劑量染氟組(500mmol/L Na F)、高劑量染氟組(5000mmol/L Na F)。觀察大鼠氟斑牙情況;用氟離子選擇電極法測定大鼠尿氟及骨氟含量;用Morris水迷宮方法測定大鼠學(xué)習(xí)記憶能力;光鏡下觀察大鼠腦組織形態(tài)學(xué)變化(HE及尼氏染色);免疫組織化學(xué)方法檢測CD11b、膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein,GFAP)的表達;酶聯(lián)免疫(Elisa)法檢測TNFa和IL-1b含量;用蛋白印跡(Western Blotting)方法檢測NF-κB p65、IκBα、Phospho-NF-κB p65和Phospho-IκB-α等因子的蛋白表達水平;用CCK8法進行細(xì)胞毒性實驗,用流式細(xì)胞術(shù)檢測染氟后THP-1細(xì)胞凋亡情況。對所獲結(jié)果作相關(guān)統(tǒng)計分析。結(jié)果:1.動物實驗:染氟組大鼠出現(xiàn)不同程度的氟斑牙,尿氟和骨氟含量明顯升高;染氟組大鼠第五天逃避潛伏期較對照組顯著延長,空間探索總次數(shù)顯著減少,差異均具有統(tǒng)計學(xué)意義(P0.05);組織形態(tài)學(xué)檢查顯示,各組大鼠腦組織HE染色未見明顯改變,但尼氏染色見染氟組大鼠腦組織神經(jīng)細(xì)胞中尼氏小體染色變淺,數(shù)量變少;免疫組化結(jié)果顯示,染氟組大鼠腦組織中CD11b及GFAP陽性表達較對照組明顯升高,差異均有統(tǒng)計學(xué)意義(P0.05);染氟組大鼠腦組織中TNFa和IL-1b含量明顯高于對照組(P0.05);染氟組大鼠腦組織中NF-κB p65及IκBα總蛋白表達均低于對照組(P0.05),但Phospho-NF-κB p65和Phospho-IκB-α蛋白表達均高于對照組;相關(guān)分析結(jié)果顯示,染氟組大鼠腦組織中Phospho-NF-κB p65表達水平與TNFa、IL-1b含量成正比,與IκBα表達成反比。2.體外培養(yǎng)細(xì)胞實驗:CCK8細(xì)胞毒性結(jié)果顯示,用500及5000mmol/L Na F培養(yǎng)細(xì)胞24、48及72小時后,500mmol/L Na F及以上濃度可明顯降低THP-1細(xì)胞存活率,呈現(xiàn)存活率與時間和劑量的負(fù)相關(guān)關(guān)系;細(xì)胞凋亡率隨著染氟時間的延長逐漸增高;低、高劑量染氟組THP-1細(xì)胞炎癥因子TNFa和IL-1b含量明顯高于對照組(P0.05);NF-κB p65、IκBα總蛋白表達均明顯低于對照組(P0.05),而Phospho-NF-κB p65和Phospho-IκB-α蛋白表達均明顯高于對照組(P0.05);相關(guān)分析結(jié)果顯示,低、高劑量染氟組THP-1細(xì)胞中Phospho-NF-κB p65表達水平與TNFa、IL-1b含量成正比,與IκBα表達成反比。結(jié)論:慢性氟中毒大鼠腦組織及過量氟暴露的體外培養(yǎng)細(xì)胞可產(chǎn)生大量的炎性因子及NF-κB信號通路激活,可能是過量的氟化鈉蓄積導(dǎo)致中樞神經(jīng)系統(tǒng)損傷的機制之一。
[Abstract]:Objective: to observe the changes of the expression of major inflammatory factors a(tumor necrosis factora1 (TNFA) and interleukin-1bsil interleukin-1b (IL-1b) in brain tissues and cultured cells of chronic fluorosis animals and to observe the inhibition of NF- 魏 B NF- 魏 B in the signal transduction pathway of nuclear factor- 魏 B(nuclear factor 魏 B and NF- 魏 B in vitro. Methods: the expression of tumor necrosis factor (TNFA) and interleukin-1b (IL-1b) in the brain of rats with chronic fluorosis were observed. Expression of protein 偽 inhibitor of nuclear factor 魏 B 偽 I 魏 B 偽 To explore its role in chronic fluorosis inflammatory brain injury mechanism. Methods: sixty male and female SD rats with chronic fluorosis were randomly divided into two groups: the control group (50 mg 路L ~ (-1) of fluoride in drinking water, added with sodium fluoride group 2), the experimental period was 10 months. Model of fluorosis cells: THP-1 cells were cultured in vitro and divided into three groups: normal control group, low dose fluoride group (500 mmol / L NaFN) and high dose fluoride group (5 000 mmol / L NAF). The fluoride content in urine and bone was determined by fluorine ion selective electrode method, and the learning and memory ability of rats was measured by Morris water maze method. The expression of CD11b, glial fibrillary acidic protein (GFAP), TNFa and IL-1b were detected by immunohistochemical method and enzyme-linked immunosorbent assay (Elisa), respectively. Western blotting method was used to detect the protein expression of factors such as Phospho-NF- 魏 B p65 and Phospho-I 魏 B- 偽, and the apoptosis of THP-1 cells after fluorosis was detected by flow cytometry. The results obtained are statistically analyzed. The result is 1: 1. Animal experiments showed that fluoride spot teeth appeared in different degrees in fluoride group, urine fluoride and bone fluorine content increased significantly, the escape latency of fluorine-exposed group on the fifth day was significantly longer than that of control group, and the total number of space exploration was significantly decreased. Histomorphological examination showed that the HE staining of brain tissue of rats in each group had no obvious change, but the staining of Nissl corpuscles in nerve cells of rats exposed to fluoride was lighter and fewer. The results of immunohistochemistry showed that the positive expression of CD11b and GFAP in the brain of rats exposed to fluoride was significantly higher than that of the control group. The contents of TNFa and IL-1b in the brain of rats exposed to fluoride were significantly higher than those of the control group (P 0.05), and the expressions of total protein of NF- 魏 B p65 and I 魏 B 偽 in the brain tissues of rats exposed to fluoride were lower than those of the control group (P 0.05), but the expression of Phospho-NF- 魏 B p65 and Phospho-I 魏 B- 偽 protein were higher than those of the control group. The results of correlation analysis showed that the expression of Phospho-NF- 魏 B p65 was directly proportional to the content of TNFa- 魏 B p65 and inversely proportional to the expression of I 魏 B 偽 in the brain of rats exposed to fluoride. The results of cell toxicity test in vitro showed that the concentration of 500 mmol / L NAF and more than 500 mmol / L NAF significantly decreased the survival rate of THP-1 cells, which showed a negative correlation with time and dose. The apoptosis rate increased gradually with the prolongation of fluorine exposure time, and decreased with the increase of fluoride exposure time. The levels of TNFa and IL-1b in THP-1 cells in high dose fluoride group were significantly higher than those in control group (P 0.05). The expression of total protein of I 魏 B 偽 was significantly lower than that of control group (P 0.05), but the expression of p65 and Phospho-I 魏 B- 偽 of Phospho-NF- 魏 B p65 and Phospho-I 魏 B- 偽 were significantly higher than that of control group (P 0.05), the correlation analysis showed that the expression of Phospho-NF- 魏 B p65 and Phospho-I 魏 B- 偽 was lower than that of control group. The expression level of Phospho-NF- 魏 B p65 in THP-1 cells exposed to high dose fluoride was directly proportional to the content of TNFa- 魏 B p65 and inversely proportional to the expression of I 魏 B 偽. Conclusion: brain tissue of rats with chronic fluorosis and cultured cells exposed to excessive fluoride can produce a large number of inflammatory factors and activation of NF- 魏 B signaling pathway, which may be one of the mechanisms of excessive accumulation of sodium fluoride leading to central nervous system damage.
【學(xué)位授予單位】：貴陽醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R595

【參考文獻】

相關(guān)期刊論文 前10條

1 劉景林,黃干川,喬廣賢,王林,劉金祥,劉全興,莫志亞,盧振明,趙永才;實驗性氟中毒大鼠病理觀察[J];中國地方病防治雜志;1998年03期

2 劉樹森,呂 嚴(yán),孫增榮,吳麗娜,盧文麗,王新偉,宋 妍;高氟區(qū)兒童智力水平調(diào)查[J];中國地方病防治雜志;2000年04期

3 王雙;朱祥宏;;高氟區(qū)兒童智力水平調(diào)查分析[J];中國地方病防治雜志;2012年01期

4 劉國艷,柴春彥,康世良;氟化物對雞甲狀腺線粒體超微結(jié)構(gòu)的影響[J];上海交通大學(xué)學(xué)報(農(nóng)業(yè)科學(xué)版);2002年03期

5 申衛(wèi)紅;彭鑫;茹松偉;宗揚勇;邵啟祥;;核轉(zhuǎn)錄因子-κB信號通路的激活與阻斷[J];生命的化學(xué);2007年03期

6 于燕妮,官志忠,張維元,姜淑芬,唐慧,楊文慶,蔣功博,劉輝;慢性氟中毒大鼠腎臟組織化學(xué)、光鏡及電鏡的變化[J];中國地方病學(xué)雜志;1988年01期

7 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

8 劉國艷,柴春彥,康世良;氟對雞甲狀腺組織結(jié)構(gòu)的影響[J];中國地方病學(xué)雜志;2001年02期

9 孫殿軍,王麗華;國際氟研究最新進展——第24屆國際氟研究協(xié)會學(xué)術(shù)會議[J];中國地方病學(xué)雜志;2002年05期

10 李達圣;安冬;何平;;貴州省燃煤型地方性氟中毒流行現(xiàn)狀調(diào)查分析[J];中國地方病學(xué)雜志;2005年06期

，

本文編號：1946994