摘要
目的评价烟酰胺腺嘌呤二核甘酸磷酸氧化酶2（NOX2）在布比卡因致神经细胞活性氧（ROS）合成中的作用。方法将SH-SY5Y细胞接种于培养板，采用随机数字表法分为4组（n＝11）：siRNA阴性对照组（NC组）、siRNA阴性对照＋布比卡因组（NC＋B组）、NOX2 siRNA组和NOX2 siRNA＋布比卡因组（NOX2 siRNA＋B组）。NC组和NOX2 siRNA组分别采用negative siRNA和NOX2 siRNA转染，随后培养基孵育24 h；NC＋B组和NOX2 siRNA＋B组分别采用negative siRNA和NOX2 siRNA转染，重新铺板，用终浓度1.5 mmol/L的布比卡因孵育3 h，更换培养基孵育至24 h。采用二氢乙锭荧光探针法检测细胞内ROS水平，采用TUNEL法确定细胞凋亡率，采用Western blot法检测活化的caspase-3和caspase-9的表达水平。结果与NC组比较，NC＋B组ROS水平、细胞凋亡率升高，活化的caspase-3和caspase-9表达上调，NOX2 siRNA＋B组ROS水平升高，活化的caspase-3和caspase-9表达上调（P 〈0.05），细胞凋亡率差异无统计学意义，NOX2 siRNA组ROS水平、细胞凋亡率差异无统计学意义（P〉0.05），活化的caspase-3和caspase-9表达上调（P〈0.05）。与NC＋B组比较，NOX2 siRNA＋B组ROS水平、细胞凋亡率降低，活化的caspase-3和caspase-9表达下调（P〈0.05）。结论NOX2参与了布比卡因诱导神经细胞ROS大量合成的病理生理机制。
ObjectiveTo evaluate the role of NOX2 in bupivacaine-induced production of reactive oxygen species （ROS） in nerve cells.
MethodsSH-SY5Y cells were seeded in culture plates and divided into 4 groups （n = 11 each） using a random number table： small interfering RNA （siRNA） negative control group （group NC）, siRNA negative control plus bupivacaine group （group NC＋ B）, NOX2 siRNA group and NOX2 siRNA plus bupivacaine group （group NOX2 siRNA＋ B）. In NC and NOX2 siRNA groups, the cells were transfected with negative siRNA and NOX2 siRNA, respectively, and then incubated in the culture medium for 24 h. In NC＋ B and NOX2 siRNA＋ B groups, cells were transfected with negative siRNA and NOX2 siRNA, respectively, new plates were used, the cells were incubated for 3 h with bupivacaine at the final concentration of 1.5 mmol/L, the culture medium was then replaced, and the cells were incubated until 24 h. The level of intracellular ROS was measured using the fluorogenic probe dihydroethidium, the cell apoptosis was determined by TUNEL, and the expression of activated caspase-3 and caspase-9 was detected using Western blot.Apoptosis rate was calculated.ResultsCompared with group NC, the level of ROS and apoptosis rate were significantly increased, and the expression of activated caspase-3 and caspase-9 was up-regulated in group NC＋ B （P〈0.05）, the level of ROS was significantly increased, and the expression of activated caspase-3 and caspase-9 was up-regulated （P〈0.05）, and no significant change was found in apoptosis rate in group NOX2 siRNA＋ B （P〉0.05）, and no significant change was found in the level of ROS or apoptosis rate （P〉0.05）, and the expression of activated caspase-3 and caspase-9 was significantly up-regulated in group NOX2 siRNA （P〈0.05）. Compared with group NC＋ B, the level of ROS and apoptosis rate were significantly decreased, and the expression of activated caspase-3 and caspase-9 was down-regulated in group NOX2 siRNA＋ B （P〈0.05）.Con
出处
《中华麻醉学杂志》
CSCD
北大核心
2017年第8期935-938,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金（81471272,81400995）
广东省自然科学基金（2014A030310075）
