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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 investigate the changes of the expression of aquaporin 8 (AQP8) in astrocytes under hypotonic pressure. Methods Primary cultured rat astrocytes were randomly divided into control group (routine culture medium) and hypotonic group (268, 254, 240 mmol / L) for 1, 3, 6, 12, 24 h, A total of 5 time points. Cell viability was measured by MTT colorimetric assay. Immunocytochemistry, immunofluorescence and Realtime-PCR were used to detect the expression of AQP8 and its mRNA. Results The morphology, cell viability and the expression of AQP8 and its mRNA in astrocytes in control group showed no significant changes at all time points (P> 0.05). In hypotonic group, astrocyte edema and cell activity decreased. The expression of AQP8 and its mRNA increased with the decrease of hypotonic degree and the prolongation of hypoosmotic time. Immunocytochemistry showed that the expression of AQP8 in astrocytes increased from hypotonic (0.2583 ± 0.0098,0.2800 ± 0.0109,0.2817 ± 0.0075) to hypotonic for 1h after hypotonic effect (268,254,240 mmol / L) (0.3850 ± 0.0109,0.4067 ± 0.0070,0.4250 ± 0.0104) (P <0.05). RealtimePCR showed that the mRNA level of AQP8 and beta-actin increased from (0.1275 ± 0.0118,0.1344 ± 0.0324,0.1675 ± 0.0078) to (3.2780 ± 0.1212, 5.4104 ± 0.1041, 8.3510 ± 0.2880). CONCLUSION: AQP8-mediated transport of water molecules leads to swelling of the astrocytes and subsequent damage to cell structure and function. AQP8 upregulation may be an important molecule that leads to the formation and development of astrocyte edema One of the mechanisms.