【摘要】：[目的]通過建立體外血腦屏障模型,評價(jià)納米氧化鋁對血腦屏障通透性的影響,初步探討血腦屏障轉(zhuǎn)運(yùn)納米氧化鋁的方式。 [方法]1.觀察納米氧化鋁顆粒(50nm)在溶液中的分散情況：(1)采用Nano-ZS90納米粒度/ZETA電位測定儀測定納米氧化鋁顆粒懸液的zeta電位及粒徑。(2)采用電鏡觀察納米氧化鋁形態(tài)及尺寸。2.建立體外血腦屏障模型：(1)原代培養(yǎng)大乳鼠腦微血管內(nèi)皮細(xì)胞、星型膠質(zhì)細(xì)胞及免疫組化檢測純度。(2)共培養(yǎng)與血腦屏障模型有效性的鑒定。3.納米氧化鋁對血腦屏障通透性的影響：(1)以0.0125mg/ml、0.025mg/ml、0.05mg/ml的納米氧化鋁對血腦屏障模型進(jìn)行染毒,比較染毒前后血腦屏障模型對蘆丁通透性變化。(2)以0.05mg/ml納米氧化鋁對血腦屏障模型進(jìn)行染毒,分別比較0、2、4、6、8h不同染毒時(shí)間對血腦屏障通透性的改變。4.血腦屏障模型轉(zhuǎn)運(yùn)納米氧化鋁方式：(1)以Cy5.5熒光標(biāo)記納米氧化鋁對血腦屏障模型進(jìn)行染毒,分別比較0、2、4、6、8、12h不同染毒時(shí)間對血腦屏障轉(zhuǎn)運(yùn)納米氧化鋁的改變。(2)以0.025mg/ml、0.05mg/mlCy5.5熒光標(biāo)記納米氧化鋁顆粒對BBB模型進(jìn)行染毒,比較不同染毒濃度對血腦屏障轉(zhuǎn)運(yùn)納米氧化鋁的改變。(3)Cy5.5熒光標(biāo)記納米氧化鋁顆粒對BBB模型進(jìn)行染毒,分別在4℃、37℃孵育,比較不同溫度對血腦屏障轉(zhuǎn)運(yùn)納米氧化鋁的改變。 [結(jié)果]1.納米氧化鋁顆粒在溶液中的分散情況：(1)納米氧化鋁顆粒粒徑分布均勻,平均粒徑為149.2nm, Zeta電位為47.6mV。(2)納米氧化鋁顆粒粒徑分布均勻,呈規(guī)則球形。2.建立體外血腦屏障模型：原代培養(yǎng)新生1-3天Wistar大鼠腦微血管內(nèi)皮細(xì)胞及星型膠質(zhì)細(xì)胞純度大于90%,BBB有效性檢測達(dá)到體外模型建立水平。3.納米氧化鋁對血腦屏障通透性的影響：(1)與0mg/ml相比,0.025mg/ml、0.05mg/ml組蘆丁吸光值明顯增加(P0.05)。(2)與0h相比,染毒2h,4h,6h,8h組,蘆丁吸光值明顯增加(P0.05)。4.血腦屏障模型轉(zhuǎn)運(yùn)納米氧化鋁方式：(1)隨時(shí)間的增加,Cy5.5熒光標(biāo)記納米氧化鋁吸光值增加(PO.05)。(2)與0.025mg/ml組比,0.05mg/ml組納米氧化鋁吸光值增加(PO.05)。(3)與4℃組比,37℃組納米氧化鋁吸光值增加(PO.05)。 [結(jié)論]1.納米氧化鋁對血腦屏障通透性的影響存在劑量效應(yīng)和時(shí)間效應(yīng)關(guān)系。 2.主動轉(zhuǎn)運(yùn)、被動擴(kuò)散方式參與了納米氧化鋁在血腦屏障的轉(zhuǎn)運(yùn)。
[Abstract]:[aim] to evaluate the effect of nano-alumina on permeability of blood-brain barrier (BBB) by establishing an in vitro blood-brain barrier (BBB) model, and to explore the transport mode of nano-Al _ 2O _ 3 by BBB. [method] 1. The dispersion of nano-alumina particles (50nm) in solution was observed. (1) the zeta potential and particle size of nano-alumina suspension were measured by Nano-ZS90 nano-particle size / ZETA potentiometer. (2) the nano-particle size was observed by electron microscope. Morphology and size of Al _ 2O _ 3 in rice-2. To establish the blood-brain barrier model in vitro: (1) Primary culture of rat brain microvascular endothelial cells, astrocytes and immunohistochemical detection of purity. (2) Co-culture and blood-brain barrier model validity. The effect of nano-alumina on the permeability of blood-brain barrier: (1) 0.0125 mg / ml, 0.025 mg / ml and 0.05 mg / ml of nano-alumina were exposed to the model of blood-brain barrier (BBB). The permeability of the blood-brain barrier model to rutin before and after exposure was compared. (2) the blood-brain barrier model was exposed to 0.05mg/ml nano-alumina, and the changes of permeability of the blood-brain barrier were compared at 0, 2, 4, 6, 8 hours after exposure. 4. The model of blood-brain barrier transport nano-alumina: (1) the blood-brain barrier model was exposed to Cy5.5 fluorescence labeled nano-alumina, compared with 0, 2, 4, 6, respectively. (2) BBB model was exposed to 0.025mg / ml and 0.05mg / ml Cy5.5 fluorescence labeled nano-alumina particles, respectively, at different time of exposure to different time. (2) BBB model was exposed to 0.025mg / ml and 0.05mg / ml Cy5.5 fluorescence labeled nano-alumina particles, respectively. The effects of different concentrations on the transport of nano-alumina at the blood-brain barrier were compared. (3) Cy5.5 fluorescent labeled nano-alumina particles were exposed to BBB model and incubated at 4 鈩，

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