纳米氧化铝对血脑屏障模型通透性影响及机制的体外研究
发布时间：2019-03-22 06:00
【摘要】：[目的]通过建立体外血脑屏障模型,评价纳米氧化铝对血脑屏障通透性的影响,初步探讨血脑屏障转运纳米氧化铝的方式。 [方法]1.观察纳米氧化铝颗粒(50nm)在溶液中的分散情况：(1)采用Nano-ZS90纳米粒度/ZETA电位测定仪测定纳米氧化铝颗粒悬液的zeta电位及粒径。(2)采用电镜观察纳米氧化铝形态及尺寸。2.建立体外血脑屏障模型：(1)原代培养大乳鼠脑微血管内皮细胞、星型胶质细胞及免疫组化检测纯度。(2)共培养与血脑屏障模型有效性的鉴定。3.纳米氧化铝对血脑屏障通透性的影响：(1)以0.0125mg/ml、0.025mg/ml、0.05mg/ml的纳米氧化铝对血脑屏障模型进行染毒,比较染毒前后血脑屏障模型对芦丁通透性变化。(2)以0.05mg/ml纳米氧化铝对血脑屏障模型进行染毒,分别比较0、2、4、6、8h不同染毒时间对血脑屏障通透性的改变。4.血脑屏障模型转运纳米氧化铝方式：(1)以Cy5.5荧光标记纳米氧化铝对血脑屏障模型进行染毒,分别比较0、2、4、6、8、12h不同染毒时间对血脑屏障转运纳米氧化铝的改变。(2)以0.025mg/ml、0.05mg/mlCy5.5荧光标记纳米氧化铝颗粒对BBB模型进行染毒,比较不同染毒浓度对血脑屏障转运纳米氧化铝的改变。(3)Cy5.5荧光标记纳米氧化铝颗粒对BBB模型进行染毒,分别在4℃、37℃孵育,比较不同温度对血脑屏障转运纳米氧化铝的改变。 [结果]1.纳米氧化铝颗粒在溶液中的分散情况：(1)纳米氧化铝颗粒粒径分布均匀,平均粒径为149.2nm, Zeta电位为47.6mV。(2)纳米氧化铝颗粒粒径分布均匀,呈规则球形。2.建立体外血脑屏障模型：原代培养新生1-3天Wistar大鼠脑微血管内皮细胞及星型胶质细胞纯度大于90%,BBB有效性检测达到体外模型建立水平。3.纳米氧化铝对血脑屏障通透性的影响：(1)与0mg/ml相比,0.025mg/ml、0.05mg/ml组芦丁吸光值明显增加(P0.05)。(2)与0h相比,染毒2h,4h,6h,8h组,芦丁吸光值明显增加(P0.05)。4.血脑屏障模型转运纳米氧化铝方式：(1)随时间的增加,Cy5.5荧光标记纳米氧化铝吸光值增加(PO.05)。(2)与0.025mg/ml组比,0.05mg/ml组纳米氧化铝吸光值增加(PO.05)。(3)与4℃组比,37℃组纳米氧化铝吸光值增加(PO.05)。 [结论]1.纳米氧化铝对血脑屏障通透性的影响存在剂量效应和时间效应关系。 2.主动转运、被动扩散方式参与了纳米氧化铝在血脑屏障的转运。
[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，
本文编号：2445310
[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，
本文编号：2445310
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