摘要
目的 研究体外培养星形胶质细胞高渗状态下水通道蛋白 4 (AQP4 ) m RNA表达的变化特点及其所起的作用。方法 取生后 2 d的新生 Wistar大鼠大脑皮质进行星形胶质细胞纯培养 ,分别用不同渗透压的高渗培养液作用于星形胶质细胞 ,建立星形胶质细胞对高渗反应的实验模型 ;采用原位杂交、乳酸脱氢酶(L DH)活性测定及图像分析等方法 ,研究体外培养星形胶质细胞对高渗液的反应和 AQP4 m RNA的表达规律。结果 32 0、333和 345 m mol/ L的高渗培养液作用于星形胶质细胞 3、6、12和 2 4 h后 ,L DH活性测定的吸光度 A值在各时间点均显著高于对照组 (P均 <0 .0 5 ) ;原位杂交分析表明 ,AQP4 m RNA表达明显增高 ,与正常组相比有显著意义 ,尤以高渗液作用 12 h时最明显 ,而且与作用时间及渗透压相关。结论 在高渗状态下 ,细胞活性明显下降 ,AQP4 m RNA表达增强。 AQP4可能在高渗性脱水的早期起重要的调节作用。
ObjectiveTo investigate the expression of aquaporin 4(AQP4) mRNA and the role in cultured rat astrocytes after exposured to hypertonic medium. MethodsRat cerebral cortical astrocytes from 2day newborn Wistar rats were seperated for the pure culture in vitro and the model of cell dehydration was established by exposed cultured astrocytes to hypertonic medium. The reactivity and rules of AQP4 mRNA expression of astrocytes to hypertonic medium were studied by in situ hybridization,the activity of lactate dehydrogenase(LDH) was detected,and image was analyzed. ResultsAfter astrocytes cultured for 3,6,12,24 hours with hypertonic medium(320,333 and 345 mmol/L),the activity of LDH showed a significant increase than that in the control group after astrocytes exposed to hypertonic medium(all P<0.05). In situ hybridization analysis confirmed that the expression of AQP4 mRNA was markedly increased,and especially it was significantly increased in hypertonic medium for 12 hours. ConclusionThe cell viability is significantly decreased and the expression of AQP4 mRNA is remarkably increased in hypertonic medium.The results suggest that AQP4 plays an important role in the early stage of the hypertonic dehydration.
出处
《中国危重病急救医学》
CAS
CSCD
2004年第4期210-213,T002,共5页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目 (3 0 0 70 2 47)
