【摘要】 联蛋白(delta-catenin)作为高表达于神经系统的黏附蛋白质,在神经系统的功能发挥中有着至关重要的作用,但其在缺血缺氧性脑病中的研究尚未见报道。本文通过体外培养原代皮层神经元,构建氧糖剥夺/再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)模型。用Western印迹、LDH等方法显示,与对照组相比,δ-联蛋白在OGD/R模型后的不同时间点(0、4、12、24和48 h)表达呈先降低后升高的趋势。在12 h表达量最低(0. 48±0. 08 vs 1. 53±0. 18,P<0. 05),在48 h表达升高到对照组水平(1. 35±0. 15 vs 1. 53±0. 18,P>0. 05)。用siRNA慢病毒干扰δ-联蛋白表达后,ELISA结果显示,和对照组相比,OGD/R后,IL-1β和IL-18升高(24. 80±1. 64 vs 12. 75±0. 87,28. 12±2. 69 vs 12. 99±1. 24,P<0. 05),但在干扰δ-联蛋白表达后,和OGD组相比,IL-1β和IL-18释放降低(12. 81±0. 78 vs 24. 80±1. 64,14. 27±1. 37 vs 28. 12±2. 69,P<0. 05)。Western印迹结果显示,AKT信号通路磷酸化位点Ser 473活化增高(1. 08±0. 04 vs 0. 85±0. 06,P<0. 05),但Thr 308位点活化无改变(1. 17±0. 06 vs 1. 11±0. 08,P>0. 05)。在siRNA慢病毒干扰并且联合使用AKT信号通路抑制剂GSK 690693后,和OGD+siRNA组相比,IL-1β和IL-18释放增高(24. 58±0. 99 vs 12. 81±0. 78,31. 62±2. 23 vs 14. 27±1. 37,P <0. 05)。上述结果显示,δ-联蛋白通过AKT信号通路调控OGD/R后的炎症反应,这可作为δ-联蛋白功能研究的新的实验依据。更多还原

【Abstract】 As a highly expressed adhesive protein in the nervous system,delta-catenin plays a crucial role in the function of the nervous system,but its role in ischemic and hypoxic encephalopathy has not been reported. In this paper, an oxygen-glucose deprivation/reperfusion(OGD/R) model was established by culturing primary cortical neurons in vitro. Western blotting and LDH showed that the expression of delta-catenin at different times of the OGD/R model(0 h,4 h,12 h,24 h,48 h),decreased first and then increased compared with the control group. The expression level decreased to the lowest level at 12 h(0. 48± 0. 08 vs 1. 53 ± 0. 18,P < 0. 05) and increased to the control level at 48 h(1. 35±0. 15 vs 1. 53±0. 18,P>0. 05). Using siRNA lentivirus interference,ELISA tests showed that compared with the control group,inflammatory factors IL-1β and IL-18 in the OGD/R model(24. 80 ±1. 64 vs 12. 75±0. 87,28. 12±2. 69 vs 12. 99±1. 24,P<0. 05) decreased after inhibition of delta-catenin(12. 81±0. 78 vs 24. 80±1. 64,14. 27±1. 37 vs 28. 12±2. 69,P<0. 05),compared with the OGD group.Moreover,compared with the OGD group,Western blotting showed that activation of phosphorylation Ser473 increased(1. 08±0. 04 vs 0. 85±0. 06,P<0. 05) while activation of phosphorylation Thr308 did not change(1. 17±0. 06 vs 1. 11±0. 08,P>0. 05). ELISA results showed that after inhibition of deltacatenin combined with the AKT pathway inhibitor GSK 690693,release of IL-1β and IL-18 increased compared with the OGD+siRNA group(24. 58±0. 99 vs 12. 81±0. 78,31. 62±2. 23 vs 14. 27±1. 37,P<0. 05). All of the data suggest that delta-catenin regulates inflammation in OGD/R through the AKT pathway,which could be a new basis for the study on the function of delta-catenin.更多还原