机构地区:[1]重庆医科大学基础医学院人体解剖学教研室,重庆400016 [2]重庆医科大学神经科学研究中心,重庆400016
出 处:《第三军医大学学报》2010年第11期1136-1140,共5页Journal of Third Military Medical University
基 金:国家自然科学基金(30470608);教育部博士培养基金(20040631002)~~
摘 要:目的探讨低渗透压作用下,星形胶质细胞水通道蛋白8(aquaporin8,AQP8)的表达变化规律。方法原代培养大鼠星形胶质细胞,随机分为对照组(常规培养液)和低渗组(268、254、240mmol/L),每组分别作用1、3、6、12、24h,共5个时间点。通过MTT比色法检测细胞活力,免疫细胞化学、免疫荧光、Realtime-PCR研究AQP8及其mRNA的表达变化。结果对照组星形胶质细胞的细胞形态、细胞活性、AQP8及其mRNA的表达在各时间点均无明显变化(P>0.05)。低渗组中,星形胶质细胞水肿,细胞活性下降;AQP8及其mRNA的表达水平随低渗程度的下调和低渗作用时间的延长而增强。低渗液(268、254、240mmol/L)作用后,免疫细胞化学显示星形胶质细胞的AQP8表达从低渗作用1h的(0.2583±0.0098,0.2800±0.0109,0.2817±0.0075)到低渗作用24h的(0.3850±0.0109,0.4067±0.0070,0.4250±0.0104)(P<0.05);RealtimePCR显示:AQP8与beta-actin的mRNA水平比从低渗作用1h的(0.1275±0.0118,0.1344±0.0324,0.1675±0.0078)上升到(3.2780±0.1212,5.4104±0.1041,8.3510±0.2880)。结论低渗作用下,AQP8介导的水分子运输可导致星形胶质细胞肿胀,继而引发细胞结构、功能的损伤;AQP8的表达上调可能是导致星形胶质细胞水肿形成和发展的重要分子机制之一。Objective To study the expression of aquaporin8 ( AQP8) in rat astrocytes under hypotonic stress. Methods Primarily cultured rat astrocytes,randomly divided into control group ( 280 to 320 mmol/L) and 3 hypotonic groups at the concentration of 268,254,and 240 mmol/L respectively,were exposed to hypotonic stress for 1,3,6,12,and 24 h,respectively. Viability of astrocytes was detected by MTT colorimetry. Expression of AQP8 at protein and mRNA levels in astrocytes was detected by immunocytochemistry,immunofluorescence,and real time reverse transcription polymerase chain reaction ( RT-PCR) ,respectively. Results No significant change was observed in morphology and viability of astrocytes as well as in expression of AQP8 and its mRNA at different time points in control group. Edema of astrocytes was observed,the viability of astrocytes was decreased,and the expression of AQP8 at protein and mRNA levels was in a negative concentration-dependent and positive time-dependent manner. Immunocytochemistry showed that the expression of AQP8 in astrocytes was 0. 258 3 ± 0. 009 8,0. 280 0 ± 0. 010 9,0. 281 7 ± 0. 007 5 respectively after astrocytes were exposed to hypotonic stress at different concentrations for 1 h and were 0. 385 0 ± 0. 010 9,0. 406 7 ± 0. 007 0,0. 425 0 ± 0. 010 4,respectively,for 24 h ( P 0. 05). RT-PCR showed that the expression of AQP8 in astrocytes was increased from 0. 258 3 ± 0. 009 8,0. 280 0 ± 0. 010 9,0. 281 7 ± 0. 007 5 in 1 h after astrocytes were exposed to hypotonic stress at different concentrations to 0. 385 ± 0. 010 9,0. 406 7 ± 0. 007 0,0. 425 0 ± 0. 010 4 in 24 h after exposure ( P 0. 05) compared to the expression of beta-actin mRNA,which was increased from 0. 127 5 ±0. 011 8,0. 134 4 ±0. 032 4,0. 167 5 ±0. 007 8 in 1 h after exposure to 3. 278 0 ± 0. 121 2,5. 410 4 ± 0. 104 1,8. 351 0 ± 0. 288 0 respectively in 24 h after exposure ( P 0. 05). ConclusionAQP8-mediated water molecule transport can lead to edema,structure and function injury of astroc
分 类 号:R322.81[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]
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