体外缺氧／复氧对星形胶质细胞AQP4表达的影响及葛根素干预的实验研究

目的观察缺氧，复氧条件下星形胶质细胞形态和AQP4mRNA的表达变化以及葛根素对其表达变化的影响，探讨脑缺血再灌注损伤与AQP4的关系以及葛根索的干预作用。方法原代培养星形胶质细胞，用5％CO2＋95％N2混合气体造成缺氧．以LDH漏出率及M1Tr降解率作为细胞受损指标，应用RT-PCR技术检验星形胶质细胞缺氧／复氧各个时间点AQP4mRNA的表达变化及葛根素的干预效果。结果体外培养的星形胶质细胞在缺氧环境下损伤不明显，随着复氧时间的延长细胞损害加重。AQP4mRNA在缺氧时表达与正常对照组无明显差异。复氧后表达升高并随时间延长呈增高趋势（P〈0．05）。葛根素干预组AQP4mRNA表达丰度与缺氧，复氧组无明显差异（P〉0．05）。结论星形胶质细胞AQP4mRNA表达变化与细胞损伤有明显的相关性，葛根素对星形胶质细胞损伤的保护作用不是通过改变AQP4的表达来实现的。

Studies of hypoxia/reoxygenation on expression of AQP4 in cultured astrocytes in vitro and the intervention effect of puerarin

Objective Observe the change of morphology of astrocytes and the expression of AQP4 mRNA and influenced by puerarin under the condition of hypoxia/reoxygenation. Probe into the relation of the ischemia/reperfusion damage of brain and AQP4 and the intervention effect of puerarin. Methods After primary culture and subculture, the astrocytes were placed in a controlled atmosphere culture chamber, into which a mixture of 95%N2 and 5%CO2 to result in cell hypoxia, regarded spilling rate of LDH and degrading rate of MTT as cellular damaged index, used RT-PCR technique to examine the expression change of AQP4 mRNA in the astrocytes at some indicated time points after hypoxia/reoxygenation and intervention effect of puerarin. Results The cultured astrocytes in vitro didn＇t damaged obviously after hypoxia, however, reoxygenation made a damage obviously. The longer time, the greater damage to the astrocytes correspondingly. The expression of AQP4 m RNA didn＇t have obvious difference to normal contrast group after 8 hours of hypoxia, but expressed rising after reoxygenation （P〈0.05）.The expression degree of AQP4 mRNA didn＇t influenced by puerarin （P〉0.05）. Conclusion There is transparent correlation between the expression change of AQP4 mRNA and damage of the astrocytes. The puerarin have protective action to the damage of astroeytes, but it doesn＇ t influenc the expression of AQP4 directly.