黄芪甲苷对氧糖剥夺复氧后星形胶质细胞上TNF-α与AQP-4表达的影响  

The influence of astragaloside Ⅳ intervention on the expression of TNF-α and AQP-4 in astrocytes after OGD/R

作　　者：李盟[1] 万志远 赵剑[1] 陈小江[1] 马全瑞[1] 刘广天[1] 黄辰[1] 文玉军[1] 秦毅[1] 

机构地区：[1]宁夏医科大学基础医学院人体解剖与组织胚胎学系,银川750004 [2]解放军513医院,酒泉735000

出　　处：《神经解剖学杂志》2016年第6期732-738,共7页Chinese Journal of Neuroanatomy

基　　金：宁夏自然科学基金(NZ14064)

摘　　要：目的:探索黄芪甲苷(astragaloside IV,ASIV)对氧糖剥夺复氧(oxygen glucose deprivation/reperfusion,OGD/R)后星形胶质细胞上肿瘤坏死因子-α(tumor necrosis facror-alpha,TNF-α)与水通道蛋-4(aquaporin-4,AQP-4)表达的影响。方法:选用第二代第1 d的星形胶质细胞。将星形胶质细胞放入缺氧培养箱(1%O_2、94%N2、5%CO_2),分别干预1、2、3、4 h,复氧24 h后用RT-PCR和Western Blot检测AQP-4和TNF-α的表达情况。将星形胶质细胞分为对照组、OGD/R组、OGD/R+0.001μmol/ml ASIV组、OGD/R+0.01μmol/ml ASIV组,用RT-PCR和免疫荧光检测AQP-4和TNF-α的表达情况。结果:RT-PCR和Western Blot结果显示:氧糖剥夺复氧后AQP-4和TNF-α的表达均增加,且AQP-4在OGD3 h时表达达到高峰,TNF-α在OGD1 h时表达达到高峰(P<0.05);OGD3h+0.001μmol/ml ASIV组和OGD3 h+0.01μmol/ml ASIV组的AQP-4基因表达水平较OGD3 h组相比均明显降低(P<0.05);与OGD3 h组相比,OGD3 h+0.001μmol/ml ASIV组的AQP-4蛋白表达水平明显降低(P<0.05);OGD1 h+0.001μmol/ml ASIV组和OGD1 h+0.01μmol/ml ASIV组的TNF-α基因表达水平与OGD1 h组相比均明显降低(P<0.05);与OGD1 h组相比,OGD1 h+0.001μmol/ml ASIV组的TNF-α蛋白表达水平明显降低(P<0.05)。结论:氧糖剥夺复氧模型可以诱导星形胶质细胞上TNF-α和AQP-4的表达,且TNF-α的表达先于AQP-4;黄芪甲苷可能是通过下调TNF-α的表达来影响AQP-4的表达,进而影响细胞水肿。Objective： To explore the influence of astragaloside IV （ASIV） intervention on the expressions of tumor necrosis facror-alpha （TNF-α） and aquaporin-4 （AQP-4） in astrocytes after OGD/R. Methods： The first day of the second generation of astrocytes were moved into an anaerobic 1% 02, 94% N2, 5% CO2 at 37℃ （1, 2, 3, 4 h）, then returned to the normal condition for 24 h. AQP-4 and TNF-α expression levels were assessed by RT-PCR and Western Blot. Astrocytes were divided into four groups： control group, OGD/R group, OGD/R ＋ 0. 001μmol/ml ASIV group, OGD/R ＋ 0.01 μmol/ml ASIV group. AQP-4 and TNF-α expression levels were assessed by RT-PCR and immunofluorescence. Results： The results of RT-PCR and Western Blot indicated that the expressions of AQP-4 and TNF-α were higher after OGD/R than those of control group, and the expressions of AQP4 at OGD3 h and TNF-α at OGD1 h were both reached the peak （P 〈 0.05 ） ; The AQP4 gene levels of the OGD3h ＋ 0.001 μmol/ml ASIV group and OGD3 h ＋ 0. 01 μmol/ml ASIV group were lower than that of the OGD3 h group （ P 〈 0.05 ） ; The AQP-4 level of the OGD3 h ＋ 0. 001 μmol/ml ASIV group was lower than that of the OGD3 h group （ P 〈 0.05 ） ; The TNF-α gene levels of the OGD1 h ＋ 0.001 μmol/ml ASIV group and the OGD1 h ＋ 0. 01 μmol/ml ASIV group were lower than that of the OGD1 h group （P 〈0.05） ; The TNF-α level of the OGD1 h ＋0. 001 μmol/ml ASIV group was lower than that of the OGD1 h group （ P 〈 0.05 ）. Conclusion： OGD/R can improve the expressions of AQP-4 and TNF-α in astrocytes, and the expression of TNF-α was prior to that of AQP-4 ; The inhibition of ASIV to AQP-4 expression can be realized by the inhibition of ASIV to TNF-α expression, and ASIV may have an effect on the edema by the ehemism.

关 键 词：水通道蛋-4 肿瘤坏死因子-α 氧糖剥夺复氧 黄芪甲苷 星形胶质细胞 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]