比较谷氨酸引起两品种大鼠培养星形胶质细胞肿胀的差异

摘要

目的：比较Wistar大鼠及SD大鼠培养星形胶质细胞（ AST）对谷氨酸所致细胞肿胀的差异。方法利用新生1 d的Wistar大鼠和SD大鼠进行AST原代及传代培养，传代培养10 d分别给予1、10 mmol／L谷氨酸孵育48 h，通过乳酸脱氢酶释放率检测细胞活性，通过胶质纤维酸性蛋白（ GFAP）免疫荧光染色、Image Pro Plus软件测量细胞周长表示细胞体积变化，通过逆转录实时荧光定量PCR法检测水通道蛋白4（AQP4） mRNA表达变化。结果谷氨酸对不同品种大鼠AST活性影响没有差异（P＞0.05）。在正常情况下，Wistar大鼠AST周长小于SD大鼠，在给予谷氨酸处理后，均明显大于SD大鼠（P＜0.05）。在给予1 mmol／L谷氨酸后，Wistar大鼠AST的AQP4 mRNA表达明显高于SD大鼠（ P＜0.05）。结论 Wistar大鼠AST对谷氨酸所致细胞肿胀较SD大鼠明显，且谷氨酸对两品种大鼠AST上AQP4表达变化的影响存在差异。%Objective To compare the differences between the cell swelling of cultured astrocytes ( AST) from Wistar and Sprague-Dawley ( SD) rats after incubation with glutamate.Methods Primary cultured AST derived from the cerebral cortex of one-day-old Wistar or SD rats were prepared.The cultured AST received 1 or 10 mmol/L glutamate treat-ment for 48 h on the tenth day after subculture.The viability of AST was determined by lactate dehydrogenase ( LDH) kit to assess the cell injury, and the perimeter of AST was measured using Image Pro Plus software after glial fibrillary acidic protein immunofluorescence staining to evaluate the astrocyte swelling.Then, the expression of aquaporin 4 ( AQP4 ) in cultured AST was detected by quantitative reverse transcription polymerase chain reaction.Results No significant differ-ence was found in the LDH release after the glutamate treatment in cultured AST from these two strains (P>0.05).The perimeter of AST from normal Wistar rats was shorter than that from SD rats, but was longer after the treatment of glutamate (P<0.05).Meanwhile, AQP4 expression in the Wistar rats was significantly higher than that from SD rats after incuba-tion with 1 mmol/L glutamate ( P<0.05 ) .Conclusions These results suggeste that cultured AST from Wistar rats are more susceptible to glutamate-induced swelling than that from SD rats, and there are differences between the effects of glu-tamate on AQP4 expression in astrocytes of Wistar and SD rats.