摘要
目的探讨重组人促红细胞生成素(recombinanthuman erythropoietin,rhEPO)预处理对氧糖剥夺并复氧培养后星形胶质细胞水肿及其水通道蛋白4(aquaporin4,AQP4)表达的影响。方法星形胶质细胞分成:正常组、模型组和rhEPO预处理组。星形胶质细胞在5 h时长的氧糖剥夺后复氧培养至0.5 h、2 h、8 h、24 h四个时间点时,用光镜观察细胞形态变化,用乳酸脱氢酶(LDH)漏出率测定反应细胞的损伤程度,用RT-PCR法及Western blot法测定AQP4的表达变化。结果光镜观察到rhEPO预处理能明显减轻氧糖剥夺后星形胶质细胞水肿的程度。氧糖剥夺后24 h时,LDH漏出率有明显增高(F=80.45,P<0.01),rhEPO预处理能明显降低(P<0.01)氧糖剥夺后LDH漏出率增高(P<0.01)的程度。AQP4的mRNA和蛋白在氧糖剥夺后的表达,各组间有明显差异(8 h时:mRNA:F=49.26,P<0.01,蛋白:F=43.13,P<0.01);与正常组相比,模型组AQP4在氧糖剥夺后短暂下降后呈上升趋势,至8 h时达峰值(均P<0.01),至24 h时未恢复正常水平;与模型组相比,rhEPO预处理组AQP4 mRNA和蛋白在氧糖剥夺后的表达变化趋势与模型组相似,但在各个时间点时的表达均明显降低(8 h时:mRNA:P<0.01,蛋白:P<0.01)。结论rhEPO预处理能明显减轻5 h氧糖剥夺后的星形胶质细胞水肿,机制可能是其通过降低氧糖剥夺后AQP4的表达水平而实现。
Objective To explore the effects of recombinant human erythropoietin（rhEPO）preconditioning on the swelling of astrocytes and the expression of AQP4 after oxygen-glucose deprivation（OGD）. Methods Astrocytes were divided into 3 groups： control group, OGD group and rhEPO preconditioning group. The effects of rhEPO on Astrocytes were investigated at 0.5, 2, 8 and 24 h after OGD. Microscope was used to observe morphological alteration of astrocytes. The cell injury was assessed by measuring LDH activity （LDH %）. The expression levels of AQP4 mRNA and protein of astrocytes were detected using RT-PCR and Western Blot, respectively. Results OGD induced obvious cell swelling and significant reduction of LDH in astrocytes whereas rhEPO preconditioning remarkably attenuated OGD-induced astrocyte swelling and LDH reduction （P 〈 0.01）. The expression levels of AQP4 mRNA and protein declined transiently at 0.5 h and then increased gradually to the peak at 8 h in OGD group（P 〈 0.05）. AQP4 mRNA and protein levels in rhEPO preconditioning group were much lower compared with OGD group at each time （P 〈 0.05）. Conclusions rhEPO preconditioning can attenuate OGD-induced swelling of astrocytes through decreasing the AQP4 expression.
出处
《中国神经精神疾病杂志》
CAS
CSCD
北大核心
2012年第1期31-35,共5页
Chinese Journal of Nervous and Mental Diseases
基金
国家自然科学基金资助项目(编号:30801182)
