Department of Anesthesiology and Critical Care, Zhongshan Hospital, Fudan University, Shanghai, PR China.
PLoS One. 2013 Jun 17;8(6):e66576. doi: 10.1371/journal.pone.0066576. Print 2013.
Nuclear factor kappa B (NF-κB) has been implicated in anesthetic preconditioning (APC) induced protection against anoxia and reoxygenation (A/R) injury. The authors hypothesized that desflurane preconditioning would induce NF-κB oscillation and prevent endothelial cells apoptosis.
A human umbilical vein endothelial cells (HUVECs) A/R injury model was used. A 30 minute desflurane treatment was initiated before anoxia. NF-κB inhibitor BAY11-7082 was administered in some experiments before desflurane preconditioning. Cells apoptosis was analyzed by flow cytometry using annexin V-fluorescein isothiocyanate staining and cell viability was evaluated by modified tertrozalium salt (MTT) assay. The cellular superoxide dismutases (SOD) activitiy were tested by water-soluble tetrazolium salt (WST-1) assay. NF-κB p65 subunit nuclear translocation was detected by immunofluorescence staining. Expression of inhibitor of NF-κB-α (IκBα), NF-κB p65 and cellular inhibitor of apoptosis 1 (c-IAP1), B-cell leukemia/lymphoma 2 (Bcl-2), cysteine containing aspartate specific protease 3 (caspases-3) and second mitochondrial-derived activator of caspase (SMAC/DIABLO) were determined by western blot.
Desflurane preconditioning caused phosphorylation and nuclear translocation of NF-κB before anoxia, on the contrary, induced the synthesis of IκBα and inhibition of NF-κB after reoxygenation. Desflurane preconditioning up-regulated the expression of c-IAP1 and Bcl-2, blocked the cleavage of caspase-3 and reduced SMAC release, and decreased the cell death of HUVECs after A/R. The protective effect was abolished by BAY11-7082 administered before desflurane.
The results demonstrated that desflurane activated NF-κB during the preconditioning period and inhibited excessive activation of NF-κB in reperfusion. And the oscillation of NF-κB induced by desflurane preconditioning finally up-regulated antiapoptotic proteins expression and protected endothelial cells against A/R.
核因子 kappa B（NF-κB）已被牵涉到麻醉预处理（APC）诱导的抗缺氧和再氧合（A/R）损伤中。作者假设，地氟醚预处理会诱导 NF-κB 振荡并防止内皮细胞凋亡。
使用人脐静脉内皮细胞（HUVEC）A/R 损伤模型。在缺氧前开始 30 分钟的地氟醚处理。在一些实验中，在进行地氟醚预处理之前，给予 NF-κB 抑制剂 BAY11-7082。通过用膜联蛋白 V-异硫氰酸荧光素染色进行流式细胞术分析细胞凋亡，通过改良 tertrozalium 盐（MTT）测定法评估细胞活力。通过水溶性四唑盐（WST-1）测定法测试细胞内超氧化物歧化酶（SOD）活性。通过免疫荧光染色检测 NF-κB p65 亚单位核易位。通过 Western blot 测定法测定抑制剂κB-α（IκBα）、NF-κB p65 和细胞凋亡抑制剂 1（c-IAP1）、B 细胞白血病/淋巴瘤 2（Bcl-2）、半胱氨酸天冬氨酸特异性蛋白酶 3（caspases-3）和第二线粒体衍生的半胱天冬氨酸蛋白酶激活剂（SMAC/DIABLO）的表达。
地氟醚预处理在缺氧前引起 NF-κB 的磷酸化和核易位，相反，在再氧合后诱导 IκBα 的合成和 NF-κB 的抑制。地氟醚预处理上调了 c-IAP1 和 Bcl-2 的表达，阻断了 caspase-3 的切割并减少了 SMAC 的释放，并减少了 HUVEC 在 A/R 后的细胞死亡。在给予地氟醚预处理之前给予 BAY11-7082 会消除保护作用。
结果表明，地氟醚在预处理期间激活 NF-κB，并抑制再灌注过程中 NF-κB 的过度激活。地氟醚预处理诱导的 NF-κB 振荡最终上调了抗凋亡蛋白的表达，并保护内皮细胞免受 A/R。
