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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 investigate the effects of astragaloside IV (ASIV) on the expression of tumor necrosis factor-alpha (TNF-α) in astrocytes after oxygen glucose deprivation / reperfusion (OGD / R) α and aquaporin-4 (AQP-4). Methods: The second-generation first-day astrocytes were selected. Astrocytes were placed in hypoxia incubator (1% O 2, 94% N 2, 5% CO 2) for 1, 2, 3, 4 h, AQP-4 and TNF-α expression. Astrocytes were divided into control group, OGD / R group, OGD / R + 0.001μmol / ml ASIV group and OGD / R + 0.01μmol / ml ASIV group. RT-PCR and immunofluorescence were used to detect AQP-4 and TNF-α expression. Results: The results of RT-PCR and Western Blot showed that the expression of AQP-4 and TNF-α increased after oxygen deprivation and the expression of AQP-4 peaked at 3 h and the expression of TNF-α reached its peak at 1 h of OGD (P <0.05). The expression of AQP-4 in OGD3h + 0.001μmol / ml ASIV group and OGD3h + 0.01μmol / ml ASIV group was significantly lower than that in OGD3h group (P <0.05) The level of AQP-4 protein in OGD3h + 0.001μmol / ml ASIV group was significantly lower than that in OGD1h + 0.001μmol / ml ASIV group and OGD1h + 0.01μmol / ml ASIV group (P <0.05) (P <0.05). Compared with OGD1 h group, the expression of TNF-α in OGD1 h + 0.001 μmol / ml ASIV group was significantly lower than that in OGD1 h group (P <0.05). Conclusion: Oxygen-deprived reoxygenation model can induce the expression of TNF-α and AQP-4 on astrocytes, and the expression of TNF-α precedes that of AQP-4; Astragaloside IV may be through down-regulating the expression of TNF-α To affect the expression of AQP-4, thereby affecting cell edema.