NADPH氧化酶在布比卡因诱发神经细胞损伤中的作用

Role of nieotinamide adenine dinucleotide phosphate hydrogen oxidase in bupivacaine-induced injury to neurons

目的评价烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶在布比卡因诱发神经细胞损伤中的作用。方法培养SH—SY5Y细胞，采用随机数字表法分为3组（n=24）：对照组（C组）用无血清培养基培养；布比卡因组（B组）用含1mmol／L布比卡因的无血清培养基孵育；加拿大麻素＋布比卡因组（A＋B组）先用含NADPH氧化酶抑制剂加拿大麻素100μmol／L的无血清培养基孵育30min，然后用1mmol／L布比卡因的无血清培养基孵育。分别于孵育2、4和6h时，每组取6孔细胞，采用MTS法检测细胞活力。于孵育4h时，每组取6孔细胞，采用流式细胞术测定ROS水平。结果与C组比较，B组孵育4和6h时细胞活力降低，孵育4h时ROS水平升高（P〈0．05）；与B组比较，A＋B组孵育细胞4和6h时细胞活力升高，孵育4h时ROS水平降低（P〈0．05）。结论NADPH氧化酶参与了布比卡因诱发的神经细胞损伤。

Objective To evaluate the role of nicotinamide adenine dinucleotide phosphate hydrogen （NADPH） oxidase in bupivacaine-induced injury to neurons. Methods SH-SY5Y cells were seeded in 96-well culture plates at a density of 5x 104 cells/well and randomly divided into 3 groups （n= 24 each） using a random number table： control group （group C）, bupivacaine group （group B）, and apoeynin （NADPH oxidase inhibitor） ＋ bupivacaine group （group A＋B）. The cells were cultured in a serum-free medium in group C. The cells were cultured in a serum-free medium containing 1 mmol/L bupivacaine in group B. In group A ＋ B, the cells were cultured for 30 min in a serum-free medium containing apocynin 100 μmol/L, and then cultured in a serum-free medium containing 1 mmol/L bupivacaine. At 2, 4 and 6 h of incubation, the cells in 6 wells of each group were selected to evaluate the cell viability by MTS assay. At 4 h of incubation, the cells in 6 wells of each group were selected to detect reactive oxygen species （ROS） level by flow cytometry. Results Compared with group C, the cell viability was significantly decreased at 4 and 6 h of incubation, and the production of ROS was increased at 4 h of incubation in group B （ P 〈 0.05）. Compared with group B, the cell viability was significantly increased at 4 and 6 h of incubation, and the production of ROS was decreased at 4 h of incubation in group B （P〈0.05）. Conclusion NADPH oxidase is involved in bupivacaine-induced injury to neurons.