本文選題：納米二氧化鈦 + 小鼠��； 參考：《蘇州大學(xué)》2013年博士論文

【摘要】：納米氧化鈦(TiO_2)是目前國際上生產(chǎn)量最大，在工業(yè)、醫(yī)藥衛(wèi)生、日用化妝品、食品以及環(huán)保方面應(yīng)用最廣泛的一種納米材料。然而隨著納米材料的廣泛應(yīng)用，包括納米TiO_2在內(nèi)的納米材料的潛在環(huán)境衛(wèi)生和職業(yè)衛(wèi)生安全越來越受到人們的高度關(guān)注，已成為研究熱點(diǎn)。2012年初，美國國家科學(xué)院國家研究理事會(huì)發(fā)布了“工程納米材料的環(huán)境、健康與安全性的研究戰(zhàn)略(A Research Strategy forEnvironmental, Health, and Safety Aspects of Engineered Nanomaterials)”報(bào)告，提出需要制定整體的研究計(jì)劃，避免納米技術(shù)快速發(fā)展帶來的潛在風(fēng)險(xiǎn)(http://www8.nationalacademies.org/onpinews/newsitem.aspx?RecordID=13347)。近年來研究，不僅證實(shí)納米TiO_2暴露可引起動(dòng)物肝、脾、肺的損傷，尤其是腎和腦的損傷，，而且闡明了相應(yīng)的損傷機(jī)制,而腎和腦這兩種器官又與生殖系統(tǒng)的功能密切相關(guān)。人們長期使用含有納米TiO_2的化妝品及長期生活或工作在暴露有納米顆粒的環(huán)境下(如生產(chǎn)車間的工人），其納米顆粒通過皮膚滲透、呼吸和消化系統(tǒng)而進(jìn)入體內(nèi)，并可能通過血液循環(huán)、血睪屏障在卵巢和睪丸中積累，進(jìn)而導(dǎo)致生殖系統(tǒng)損傷。但納米TiO_2暴露對(duì)生殖系統(tǒng)的損傷及其機(jī)制研究尚不夠深入。因此開展納米TiO_2生殖系統(tǒng)毒性的研究具有非常重要的理論意義和實(shí)際意義。鑒于此，本論文以10mg/kg BW的納米TiO_2（銳鈦型5nm）連續(xù)灌胃處理雌性和雄性小鼠90天，對(duì)其生殖系統(tǒng)的損傷進(jìn)行了研究；同時(shí)結(jié)合基因芯片技術(shù)深入研究了生殖系統(tǒng)損傷多基因共同作用的分子機(jī)制。本研究可為納米材料長期暴露引起生殖系統(tǒng)的毒性評(píng)估提供重要的理論依據(jù)。 論文結(jié)果如下： (1)用納米TiO_2(10mg/kg)對(duì)雌性小鼠持續(xù)灌胃90天后，研究了雌性小鼠卵巢的損傷和卵巢基因表達(dá)特征。結(jié)果顯示納米TiO_2顆粒能夠進(jìn)入卵巢中積累并且導(dǎo)致卵巢的損傷如卵巢萎縮，初級(jí)卵泡和次級(jí)卵泡發(fā)育障礙，卵巢細(xì)胞的不規(guī)則排列和卵泡腔的形狀不規(guī)則。納米TiO_2顆粒也可進(jìn)入卵巢細(xì)胞甚至細(xì)胞核，進(jìn)而導(dǎo)致細(xì)胞凋亡。納米TiO_2顆粒暴露造成了卵巢組織鈣、鈉、鉀和鋅金屬元素的含量增加，而鎂、銅和鐵金屬元素的含量下降。納米TiO_2顆粒暴露導(dǎo)致了性激素的不平衡，如血清孕酮、促黃體生成素、睪酮和促卵泡激素的水平降低，雌二醇的濃度增加，進(jìn)而引起生育力下降，如交配率、懷孕率、產(chǎn)仔數(shù)都顯著下降及子代生長發(fā)育減慢。同時(shí)納米TiO_2顆粒暴露引起了卵巢嚴(yán)重的氧化性損傷，如ROS大量積累和DNA氧化水平增加。微陣列分析顯示，納米TiO_2處理的小鼠卵巢組織中有223個(gè)已知功能基因上調(diào)(diffscore≥13, p0.05)，65個(gè)已知功能基因下調(diào)(diffscore-13, p0.05)。在本研究中，卵巢的差異基因的功能分別涉及細(xì)胞分化、免疫炎癥反應(yīng)、離子運(yùn)輸、代謝過程、氧化應(yīng)激反應(yīng)、內(nèi)固醇代謝、凋亡、信號(hào)轉(zhuǎn)導(dǎo)、信號(hào)通路、轉(zhuǎn)錄、運(yùn)輸?shù)�。尤其是與內(nèi)固醇和性激素代謝過程有關(guān)的Akr1c18、Cyp17a1和Lgmn顯著上調(diào)，這些基因的過表達(dá)引起雌二醇濃度上升，以及孕酮、促黃體生成素和睪酮濃度下降，進(jìn)而抑制卵泡發(fā)育和降低雌性小鼠的生育力。另外，納米TiO_2顆粒暴露后導(dǎo)致卵巢Cyp17a1的表達(dá)顯著增加也可促進(jìn)雌二醇的生物合成。 (2)分析了10mg/kg納米TiO_2鈦顆粒持續(xù)90天灌胃雄性小鼠后生殖毒性和基因表達(dá)譜變化。結(jié)果表明納米TiO_2顆粒能夠通過血睪屏障在睪丸中沉積，并且導(dǎo)致睪丸的損傷和細(xì)胞凋亡，如生精管空泡化、生精層的厚度減小，睪丸細(xì)胞的不規(guī)則排列和支持細(xì)胞的空泡化；附睪尾部的精子發(fā)生異常的變化，如總的精子濃度和精子活力下降，精子畸形率顯著增加，精子損傷嚴(yán)重如頭尾斷裂；雄性小鼠的生育力顯著下降，如低的交配率、妊娠率和少的產(chǎn)仔數(shù)。納米TiO_2的暴露導(dǎo)致睪丸組織中鈣和鐵的含量下降，鋅含量顯著增加。同樣，納米TiO_2暴露也造成性激素的不平衡，如雌二醇和孕酮顯著增加，而促卵泡激素、促黃體生成素和睪酮水平顯著下降。微陣列分析顯示與對(duì)照比較，納米TiO_2處理組的小鼠睪丸組織有155個(gè)基因明顯上調(diào)(diffscore≥13, p0.05)，100個(gè)基因明顯下調(diào)(diffscore-13,p0.05)。對(duì)已知功能的差異基因分類，表明差異基因的功能涉及的范疇有精子發(fā)生、內(nèi)固醇代謝過程、凋亡、免疫、細(xì)胞分化、氧化應(yīng)激反應(yīng)、氧化還原活性、激素活性、離子運(yùn)輸?shù)�。尤其是Spata19、Tdrd6和Tnp2表達(dá)的顯著下調(diào)直接與精子發(fā)生的抑制有關(guān)，而Cyp2e1、Mvd、Sc4mol和Srd5a2與內(nèi)固醇代謝有關(guān),Cyp2e1表達(dá)下調(diào)，Mvd、Sc4mol和Srd5a2表達(dá)上調(diào)。
[Abstract]:Nanoscale titanium oxide (TiO_2) is the most widely used nano material in the field of industry, medicine and health, daily cosmetics, food and environmental protection. However, with the wide application of nanomaterials, the potential of nanomaterials, including nano TiO_2, is becoming more and more popular in environmental hygiene and occupational health. At the beginning of.2012, the National Research Council of the National Academy of Sciences issued the "A Research Strategy forEnvironmental, Health, and Safety Aspects of Engineered Nanomaterials) report, and proposed the need to develop the overall research. To avoid the potential risk (http://www8.nationalacademies.org/onpinews/newsitem.aspx? RecordID=13347) from the rapid development of Nanotechnology (http://www8.nationalacademies.org/onpinews/newsitem.aspx? RecordID=13347). In recent years, it has been studied not only that nano TiO_2 exposure can cause damage to animal liver, spleen and lung, especially kidney and brain damage, but also elucidate the corresponding damage mechanism, while the kidney and brain are two The organ is also closely related to the function of the reproductive system. People have long used cosmetics containing nano TiO_2 and long life or work in the environment of exposed nanoparticles, such as workers in the production workshop, whose nanoparticles enter the body through the skin infiltration, respiration and digestive systems, and may circulate through the blood and the blood testis barrier in the body. The accumulation of the ovary and testicle leads to the damage of the reproductive system, but the research on the damage and mechanism of the reproductive system by nano TiO_2 exposure is not enough. Therefore, it is of great theoretical and practical significance to study the toxicity of the reproductive system of the nano TiO_2 reproductive system. In view of this, this article uses the 10mg/kg BW nano TiO_2 (anatase 5nm) continuously. The female and male mice were treated with gavage for 90 days, and the damage of their reproductive system was studied. At the same time, the molecular mechanism of the multigene interaction of reproductive system injury was studied by gene chip technology. This study could provide important theoretical basis for the assessment of the toxicity of the reproductive system by the long-term exposure of nanomaterials.
The results of the paper are as follows:
(1) the ovarian damage and ovarian gene expression in female mice were studied after 90 days after continuous gavage of female mice with TiO_2 (10mg/kg). The results showed that the nano TiO_2 particles could accumulate in the ovary and cause ovarian damage, such as ovarian atrophy, primary follicle and secondary follicle development disorder, irregular arrangement of ovarian cells and the abnormal arrangement of ovarian cells. The shape of the follicle cavity is irregular. Nanoscale TiO_2 particles can also enter ovarian cells and even nuclei and lead to cell apoptosis. The exposure to nano TiO_2 particles causes the increase in the content of calcium, sodium, potassium and zinc metal elements in the ovarian tissue, while the content of magnesium, copper and iron elements is decreased. The exposure of nano TiO_2 particles leads to the imbalance of sex hormones. Such as serum progesterone, luteinizing hormone, testosterone and follicle stimulating hormone levels, the increase of estradiol, and the decrease of fertility, such as mating rate, pregnancy rate, litter size and progeny growth slowing. Meanwhile, the nano TiO_2 particle exposure causes severe oxidative damage to the ovary, such as the accumulation of ROS and DNA oxygen. Microarray analysis showed that there were 223 known functional genes up regulation (diffscore > 13, P0.05) and 65 known functional genes down regulation (diffscore-13, P0.05) in the mouse ovarian tissue treated with nanoscale TiO_2. In this study, the functional differentiation of the ovarian differential genes involved cell differentiation, immuno inflammatory response, ion transport, metabolism. Process, oxidative stress response, metabolism of sterol, apoptosis, signal transduction, signal transduction, signaling, transcription, transport, etc., especially Akr1c18, Cyp17a1 and Lgmn associated with steroid and sex hormone metabolism, the overexpression of these genes increases the concentration of estradiol, and progesterone, luteinizing hormone and testosterone, and then inhibition of eggs. Bubbles develop and reduce the fertility of female mice. In addition, after exposure to nano TiO_2 particles, the expression of Cyp17a1 in the ovary is increased significantly and the biosynthesis of estradiol can also be promoted.
(2) the reproductive toxicity and gene expression profiles of 10mg/kg nanoscale TiO_2 titanium particles were analyzed after 90 days of gastric perfusion in male mice. The results showed that the nano TiO_2 particles could be deposited through the blood testis barrier in the testis, and caused the damage and apoptosis of the testis, such as the vacuolation of spermatogenic tube, the decrease of the thickness of spermatogenic layer, and the irregular row of the testicular cells. The vacuolization of columns and supporting cells; abnormal changes in spermatogenesis in the tail of the epididymis, such as the total sperm concentration and sperm motility, a significant increase in sperm abnormality, and severe sperm damage such as the head and tail fracture; the fertility of male mice decreased significantly, such as low mating rate, pregnancy rate and litter size. The exposure to nano TiO_2 led to the testicles. The content of calcium and iron in the tissue decreased and the zinc content increased significantly. Similarly, TiO_2 exposure also resulted in the imbalance of sex hormones, such as estradiol and progesterone, while follicle stimulating hormone, luteinizing hormone and testosterone levels decreased significantly. Microarray analysis showed that there were 155 mice in the nano TiO_2 treatment group compared with the control group. The genes were significantly up-regulated (diffscore > 13, P0.05), and 100 genes were obviously down regulated (diffscore-13, P0.05). The differential gene classification of known functions indicated that the function of the differential genes involved spermatogenesis, sterol metabolic process, apoptosis, immunization, cell differentiation, oxidative stress reaction, redox activity, hormone activity, ion transport, etc. In particular, the significant down-regulation of Spata19, Tdrd6 and Tnp2 is directly related to the inhibition of spermatogenesis, while Cyp2e1, Mvd, Sc4mol and Srd5a2 are related to the metabolism of sterol, the expression of Cyp2e1 is down, Mvd, Sc4mol and Srd5a2 are up-regulated.

【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 薛猛;朱融融;孫曉宇;汪世龍;;納米二氧化鈦的發(fā)育毒性研究[J];材料導(dǎo)報(bào);2009年04期

2 侯娟;萬旭英;王飛;許桂鳳;劉珍;張?zhí)鞂?;納米級(jí)二氧化鈦對(duì)體外大鼠腔前卵泡生長發(fā)育成熟的影響[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2009年08期

3 陸荔,馬明,張宇,唐萌,顧寧;納米材料生物安全性研究進(jìn)展[J];東南大學(xué)學(xué)報(bào)(自然科學(xué)版);2004年05期

4 王燕;康現(xiàn)江;丁士文;穆淑梅;王宇;曹輝彩;;納米二氧化鈦對(duì)小鼠肝腎的影響[J];環(huán)境與健康雜志;2008年02期

5 汪冰,豐偉悅,趙宇亮,邢更妹,柴之芳,王海芳,賈光;納米材料生物效應(yīng)及其毒理學(xué)研究進(jìn)展[J];中國科學(xué)(B輯 化學(xué));2005年01期

6 羊富光;湯瑩;于洋;范曉燕;許珊;沈亞峰;劉關(guān)省;楊勇驥;;二氧化鈦納米顆粒對(duì)人角質(zhì)形成細(xì)胞超微結(jié)構(gòu)的影響及毒性效應(yīng)[J];解剖學(xué)雜志;2009年02期

7 應(yīng)賢平;;納米顆粒對(duì)大氣環(huán)境和人類健康的影響[J];環(huán)境與職業(yè)醫(yī)學(xué);2006年01期

8 郭利利;劉曉慧;秦定霞;高利;張紅梅;劉嘉茵;崔毓桂;;納米二氧化鈦對(duì)雄性小鼠生殖系統(tǒng)的影響[J];中華男科學(xué)雜志;2009年06期

9 王海濤,楊祥良,徐輝碧;活性氧的信號(hào)分子作用[J];生命的化學(xué);2001年01期

10 朱融融,汪世龍,姚思德;納米二氧化鈦的生物學(xué)效應(yīng)[J];生命的化學(xué);2005年04期

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