吡拉格雷钠对氧糖剥夺／复氧后星形胶质细胞水肿及 AQP4表达的影响

摘要

目的：探讨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（AQP4）表达的变化以及吡拉格雷（TSF）对其表达变化的影响。方法体外培养原代星形胶质细胞，建立氧糖剥夺／复氧细胞水肿模型，并随机分为正常组、氧糖剥夺／复氧损伤（OGD／Reox）组、奥扎格雷钠（Ozagrel）阳性对照组和 TSF 治疗组，在氧糖剥夺／复氧损伤后6、12、24和48 h 4个时间点测定细胞体积变化，乳酸脱氢酶（LDH）漏出率、MTT 法检测细胞损伤及存活情况，Western blot 法检测各组细胞膜 AQP4蛋白的表达水平，RT-PCR 法检测各组细胞AQP4 mRNA 的转录水平。结果星形胶质细胞经缺氧再复氧处理，随着复氧时间的延长细胞损害加重（P ＜0.05，P ＜0.01）；TSF 治疗组的细胞体积明显低于 OGD／Reox 损伤组（P ＜0.05）；给予 TSF 治疗后细胞在氧糖剥夺／复氧后的LDH 漏出率较 OGD／Reox 损伤组明显降低（P ＜0.05）；TSF治疗组与单纯氧糖剥夺／复氧相比较 MTT 检测细胞活力明显升高（P ＜0.05）；给予 TSF 治疗组，AQP4表达明显降低（P＜0.05）；与 Ozagrel 相比较差异无显著性。Western blot 及RT-PCR 检测蛋白及 mRNA 的表达水平，给予 TSF 治疗组的AQP4蛋白及 mRNA 表达均明显降低（P ＜0.05）。结论TSF 治疗能明显减轻体外氧糖剥夺／复氧损伤引起的星形胶质细胞水肿，可能是通过降低氧糖剥夺后 AQP4的表达水平而实现。%Aim To explore the effects of tetrameth-ylpyrazine-2′-O-sodium ferulate (TSF)on the swelling of astrocytes and the expression of AQP4 after oxygen gl-ucose deprivatio /reoxygenation(OGD /Reox).Methods Astrocytes were divided into 4 groups:control group, OGD /Reox group,Ozagrel group and TSF group.The effects of TSF on astrocytes were investigated 6,1 2,24 and 48 h after OGD /Reox.The cell injury was assessed by measuring LDH activity and MTT.The expression levels of AQP4 protein of astrocytes were detected u-sing Western blot.Results OGD /Reox induced obvi-ous cell swelling and significant reduction of LDH in astrocytes whereas TSF remarkably attenuated OGD-in-duced astrocyte swelling and LDH reduction (P 0.05 ).Conclusion TSF can attenuate OGD-induced swelling of astrocytes through decreasing the AQP4 expression.