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目的:研究星形胶质细胞CTX TNA2在氧糖剥夺(oxygen-glucose deprivation,OGD)后N-myc downstream regulated gene 2(NDRG2)和其磷酸化分子p-NDRG2(Thr-348)的表达变化,明确NDRG2的磷酸化修饰是否对星形胶质细胞存活产生影响。方法:将CTX TNA2细胞进行3 h OGD处理,复氧复糖后于0 h,2 h,6 h,12 h,24 h,通过Western Blot检测NDRG2和p-NDRG2(Thr-348)水平的变化。再通过感染慢病毒或给予Akt改变pNDRG2(Thr-348),研究不同水平的p-NDRG2(Thr-348)对CTX TNA2细胞的存活是否产生影响。结果:(1)CTX TNA2细胞在OGD后2 h NDRG2(Thr-348)磷酸化水平达到峰值,以后逐渐下降,24 h后会恢复到OGD后0 h的水平;(2)抑制p-NDRG2(Thr-348),降低CTX TNA2细胞活力(P<0.05),增加乳酸脱氢酶(lactate dehydrogenase,LDH)释放(P<0.05);(3)促进p-NDRG2(Thr-348),提高CTX TNA2细胞活力(P<0.05),减少LDH释放(P<0.05);结论:星形胶质细胞CTX TNA2细胞在OGD后NDRG2磷酸化水平发生明显变化。促进NDRG2磷酸化,有助于细胞存活;抑制NDRG2磷酸化,加重细胞凋亡。
Objective: To investigate the expression of NDRG2 and its phosphorylation molecule p-NDRG2 (Thr-348) in astrocyte CTX TNA2 after oxygen glucose deprivation (OGD) It is clear whether the phosphorylation of NDRG2 affects the survival of astrocytes. Methods: CTX TNA2 cells were treated with OGD for 3 h. The changes of NDRG2 and p-NDRG2 (Thr-348) levels were detected by Western Blot at 0 h, 2 h, 6 h, 12 h and 24 h after reoxygenation / . The effect of varying levels of p-NDRG2 (Thr-348) on the survival of CTX TNA2 cells was also examined by infection with lentiviruses or alteration of pNDRG2 (Thr-348) with Akt. Results: (1) The phosphorylation level of NDRG2 (Thr-348) peaked at 2 h after OGD in CTX TNA2 cells and then gradually decreased after 24 hours. The levels of p-NDRG2 Thr-348), and decreased the viability of CTX TNA2 cells (P <0.05), and increased the release of lactate dehydrogenase (LDH) (P <0.05) (P <0.05), and decreased the release of LDH (P <0.05). Conclusion: The level of NDRG2 phosphorylation in astrocyte CTX TNA2 cells changed significantly after OGD. Promote NDRG2 phosphorylation, contribute to cell survival; inhibit NDRG2 phosphorylation, increase cell apoptosis.