摘要 目的 探讨沉默信号调节因子1（SIRT1）/细胞核因子-κB（NF-κB）信号通路在氧糖剥夺再灌注诱导海马神经元损伤中的作用。 方法 通过SIRT1过表达慢病毒和RNAi技术调控小鼠海马神经元SIRT1的表达后建立氧糖剥夺再灌注损伤模型，实验设置七组：对照组（Control组）、氧糖剥夺再灌注损伤模型组（OGD组）、OGD+转染SIRT1-siRNA质粒组（OGD+SIRT1-siRNA组）、OGD+转染空载质粒组（OGD+NC-siRNA组）、OGD+感染过表达SIRT1慢病毒组（OGD+SIRT1上调组）、OGD+感染空载慢病毒组（OGD+Vector组）及OGD+SIRT1上调+ SIRT1-siRNA组。对照组在37 ℃常氧条件下培养，其余六组在经过相应处理后给予氧糖剥夺处理9 h及再灌注处理24 h，再灌注处理结束后采用CCK-8法测定细胞活力、化学比色法测定乳酸脱氢酶（LDH）释放量、流式细胞术检测细胞凋亡率，荧光定量PCR法检测SIRT1、NF-κB的mRNA表达量，Western blot检测SIRT1、NF-κB、IκBα、Bcl-2、Bax、Cleaved caspase-3、β-actin、TBP的表达量。 结果 与Control组比较，OGD组细胞活力（%：100 vs. 53.83±1.88)、SIRT1蛋白表达量(0.62±0.06 vs. 0.33±0.03)，IκBα蛋白表达量(1.01±0.06 vs. 0.42±0.03)、Bcl-2/Bax比值(2.64±0.34 vs. 0.58±0.06)明显降低（P＜0.05），LDH释放量(U/mL：1.12±0.17 vs. 2.76±0.23)，细胞凋亡率(%：8.46±1.77 vs. 29.58±1.84)，NF-κB蛋白表达量(0.23±0.03 vs. 0.63±0.03)，Cleaved caspase-3蛋白表达量(0.19±0.03 vs. 0.82±0.04)明显升高（P＜0.05）；与OGD组和OGD+Vector组比较，OGD+SIRT1上调组细胞活力(%：53.83±1.88、50.33±3.83 vs. 72.77±1.80)、SIRT1表达量(0.63±0.03、0.58±0.04 vs. 1.02±0.05)、IκBα表达量(0.42±0.03、0.38±0.04 vs. 0.62±0.04)及 Bcl-2/Bax比值(0.58±0.06、0.55±0.04 vs.1.41±0.16)明显升高（P＜0.05），LDH释放量(U /mL：2.76±0.23、2.91±0.25 vs.1.95±0.20)、细胞凋亡率(%：29.58±1.84、28.87±2.91 vs. 20.61±2.81)%、NF-κB表达量(0.63±0.03、0.58±0.04 vs.0.35±0.04)、Cleaved caspase-3表达量(0.82±0.04、0.71±0.05 vs. 0.46±0.04)明显降低（P＜0.05）；与OGD组和OGD+NC-siRNA组比较，OGD+SIRT1-siRNA组细胞活力(%：53.83±1.88、49.23±3.12 vs. 34.86±3.17)、SIRT1表达量(0.33±0.03、0.32±0.04 vs. 0.18±0.04)、IκBα表达量(0.42±0.03、0.46±0.05 vs. 0.27±0.04)及Bcl-2/Bax比值(0.580.06、0.62±0.05 vs. 0.15±0.02)明显降低（P＜0.05），LDH释放量(U/mL：2.76±0.23、2.78±0.24 vs. 3.51±0.31 )、细胞凋亡率(%：29.58±1.84、28.52±2.08 vs. 37.03±3.19)、NF-κB表达量(0.63±0.03、0.64±0.04 vs. 0.93±0.05)、Cleaved caspase-3表达量(0.82±0.04、0.85±0.05 vs. 1.12±0.13)明显升高（P＜0.05）。结论 SIRT1/NF-κB通路参与了氧糖剥夺再灌注诱导小鼠海马神经元损伤过程。
姚鹏，陈勇，徐国海. SIRT1/NF-κB通路在氧糖剥夺再灌注诱导小鼠海马神经元损伤中的作用[J]. 中国急救医学, 2019, 39(4): 382-387.
Yao Peng, Chen Yong, Xu Guo-hai. The role of SIRT1/NF-κB pathway in injury of mouse hippocampal neurons induced by oxygen glucose deprivation and reperfusion. Chinese Journal of Critical Care Medicine, 2019, 39(4): 382-387.
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