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  

Effects of tetramethylpyrazine-2'-O-sodium ferulate on swelling of astrocytes and expression of AQP4 after oxygen-glucose deprivation/reoxygenation

作　　者：许淑红[1] 尹茂山 吴玉林[1] 

机构地区：[1]中国药科大学药学院,江苏南京211198 [2]济南大学山东省医学科学院医学与生命科学学院,山东济南250200 [3]山东省医学科学院药物研究所,山东济南250062

出　　处：《中国药理学通报》2015年第12期1661-1667,共7页Chinese Pharmacological Bulletin

基　　金：国家重大专项基金资助项目(No 2009ZX09103-123)

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 m RNA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m RNA的表达水平,给予TSF治疗组的AQP4蛋白及m RNA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 tetramethylpyrazine-2'-O-sodium ferulate( TSF) on the swelling ofastrocytes and the expression of AQP4 after oxygen glucose 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 6,12,24 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 using Western blot. Results OGD / Reox induced obvious cell swelling and significant reduction of LDH in astrocytes whereas TSF remarkably attenuated OGD-induced astrocyte swelling and LDH reduction( P <0. 01). The expression levels of AQP4 protein increased gradually to the peak at 48 h in OGD / Reoxgroup( P < 0. 05). AQP4 protein levels in TSF group were much lower compared with OGD / Reox group at each time point( P < 0. 05). And there was no significant difference between TSF group and OGD / Reox group( P > 0. 05). Conclusion TSF can attenuate OGD-induced swelling of astrocytes through decreasing the AQP4 expression.

关 键 词：吡拉格雷钠 星形胶质细胞 氧糖剥夺/复氧 细胞体积 乳酸脱氢酶漏出率 细胞水肿 水通道蛋白4 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]