本文選題：納米二氧化硅顆粒 + 血管內(nèi)皮細(xì)胞�。� 參考：《華中科技大學(xué)》2013年碩士論文

【摘要】：目的:納米材料的職業(yè)安全和健康問(wèn)題是很復(fù)雜的。納米材料雖然粒徑小卻有較大的表面積，導(dǎo)致其與相同成分較大尺寸的顆粒物相比，物理、化學(xué)、生物學(xué)性質(zhì)發(fā)生較大的變化，這些理化性質(zhì)極有可能會(huì)影響納米材料以下幾方面的能力，到達(dá)肺氣體交換區(qū)域、自肺進(jìn)入運(yùn)輸至整個(gè)機(jī)體、穿透皮膚屏障、跨越細(xì)胞膜，甚至在分子水平的反應(yīng)等等。初步的一些研究結(jié)果表明，微米級(jí)物質(zhì)的安全性評(píng)價(jià)結(jié)果可能并不不適用于納米級(jí)物質(zhì)，因此納米尺度材料的生物安全性問(wèn)題得到人們的廣泛關(guān)注。故本研究選擇比較常見(jiàn)的典型納米顆粒（SiO2顆粒）作為染塵顆粒，通過(guò)體外實(shí)驗(yàn)對(duì)納米顆粒物心血管毒性的劑量和時(shí)間效應(yīng)進(jìn)行研究，為納米級(jí)化學(xué)物的心血管安全性評(píng)價(jià)提供進(jìn)一步的實(shí)驗(yàn)依據(jù)。 方法:選擇nano-SiO2作為染塵顆粒物，將人臍靜脈內(nèi)皮細(xì)胞（HUVECs）作為受試細(xì)胞，不同劑量（0、5、10、25、50、100、200μg/ml）的顆粒物和人臍靜脈內(nèi)皮細(xì)胞共培養(yǎng)不同時(shí)間6、12、24、48h，用CCK-8方法檢測(cè)細(xì)胞存活率，計(jì)算半數(shù)致死量，確定后續(xù)實(shí)驗(yàn)的染塵劑量為0、5、10、20、40μg/ml。將上述劑量（0、5、10、20、40μg/ml）的nano-SiO2顆粒與HUVECs共培養(yǎng)6、12、24、48h。用相應(yīng)試劑盒測(cè)定細(xì)胞培養(yǎng)液上清中乳酸脫氫酶（LDH）活性、總超氧化物歧化酶（SOD）活性、腫瘤壞死因子-α（TNF-α）釋放量、白介素-6（IL-6）的釋放量。 結(jié)果:細(xì)胞存活率隨著nano-SiO2染塵劑量升高和作用時(shí)間的延長(zhǎng)而逐漸降低。如果作用時(shí)間短，細(xì)胞存活率在較高劑量開(kāi)始明顯降低；若作用時(shí)間長(zhǎng)，細(xì)胞存活率在低劑量即明顯下降。 細(xì)胞培養(yǎng)液中LDH活性、TNF-α釋放量、IL-6釋放量基本上隨著染塵劑量的升高和作用時(shí)間的延長(zhǎng)而逐漸升高。 隨著作用的時(shí)間延長(zhǎng)，細(xì)胞因子TNF-α和IL-6開(kāi)始出現(xiàn)明顯變化的劑量有逐漸降低的趨勢(shì)，反之亦然；LDH活性變化的劑量則未見(jiàn)這個(gè)趨勢(shì)，明顯變化的劑量始終在10μg/ml。 SOD活力的變化與上述指標(biāo)不同。在6、12h時(shí)間點(diǎn)，隨著染塵劑量增加，SOD活力呈升高趨勢(shì)，在24h時(shí)間點(diǎn)，，SOD活力呈先增高后降低的改變，在48h時(shí)間點(diǎn)，SOD活力呈降低趨勢(shì)。隨著作用時(shí)間的延長(zhǎng)，SOD活力呈現(xiàn)先升高后降低的趨勢(shì)。 結(jié)論: Nano-SiO2能夠?qū)UVECs產(chǎn)生一定的毒性效應(yīng)，且對(duì)細(xì)胞的毒性效應(yīng)存在明顯的劑量和時(shí)間依賴性。
[Abstract]:Objective: occupational safety and health problems of nanomaterials are complex. Nanomaterials have a large surface area, although they are small in size, resulting in great changes in physical, chemical and biological properties compared with those of larger particles of the same composition. These physical and chemical properties are likely to affect the ability of nanomaterials to reach the lung gas exchange region, transport from the lung to the whole body, penetrate the skin barrier, cross the cell membrane, and even react at the molecular level, etc. Some preliminary research results show that the safety evaluation results of micron materials may not be applicable to nanoscale materials, so the biosafety of nanoscale materials has been paid more and more attention. Therefore, in this study, typical nano-particles (SiO2) were selected as dust particles, and the dose and time effects of cardiovascular toxicity of nanoparticles were studied in vitro. To provide further experimental basis for cardiovascular safety evaluation of nano-scale chemicals. Methods: human umbilical vein endothelial cells (HUVECs) were selected as dust-contaminated particles and human umbilical vein endothelial cells (HUVECs) were co-cultured with human umbilical vein endothelial cells (HUVECs) at different doses for 6 ~ 12 ~ 24 ~ (24) h for different time. The survival rate of cells was measured by CCK-8 method, and the lethal dose of HUVECs was calculated by 50 渭 g / ml of human umbilical vein endothelial cells (HUVECs). The dose of dust in the follow-up experiment was determined to be 0 ~ 5 ~ 10 ~ 10 ~ 20 渭 g 路ml ~ (-1) 路L ~ (-1). The above dosages of nano-SiO2 particles were co-cultured with HUVECs at a dose of 10 ~ 10 渭 g / ml and 60 渭 g / ml, respectively, and then cultured for 48 h with 6o _ (12) O _ (24) O _ (24) 渭 g / ml. The activity of lactate dehydrogenase (LDH), the activity of total superoxide dismutase (SOD), the release of tumor necrosis factor- 偽 (TNF- 偽) and the release of interleukin-6 (IL-6) in the supernatant of cell culture medium were measured by the corresponding kit. Results: the cell survival rate decreased gradually with the increase of nano-SiO2 dust exposure dose and the prolongation of the time of action. If the time was short, the cell survival rate began to decrease at a higher dose, and if the time was long, the cell survival rate decreased significantly at a low dose. The release of TNF- 偽 and IL-6 in the cell culture medium increased gradually with the increase of dust exposure dose and the prolongation of the time of action. The dosages of TNF- 偽 and IL-6 began to change gradually with the prolongation of the time of action, but the dose of the change of activity of TNF- 偽 and IL-6 was not seen at the same time, and the dose of obvious change was always at 10 渭 g / ml. The change of SOD activity was different from the above indexes. At 6h / 12h, the SOD activity increased with the increase of dust exposure dose, increased first and then decreased at 24h, and decreased at 48h. The activity of SOD increased first and then decreased with the prolongation of action time. Conclusion: Nano-SiO2 can produce a certain toxic effect on HUVECs, and its cytotoxic effect is dose and time dependent.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

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