异丙酚对胎鼠离体海马神经元钙离子浓度和 NF-κB 活性的影响  

Effects of propofol on intracellular calcium ion concentration and NF-κB activity in hippocampal neurons of fetal rats in vitro

作　　者：陈静[1] 利莉[1] 梁羽冰[1] 谢玉波[1] 

机构地区：[1]广西医科大学第一附属医院麻醉科,南宁市530021

出　　处：《中华麻醉学杂志》2014年第3期286-289,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81060277,81373498）;广西科学研究与技术开发计划项目（桂科攻1355005-4-2）;广西高校科学技术研究项目（2013ZD014）

摘　　要：目的：评价异丙酚对体外培养的胎鼠海马神经元内钙离子浓度（[Ca2＋]i ）和NF-κB活性的影响。方法孕16～18 d SD大鼠拉颈处死，剖腹取胎鼠，分离海马神经元。将神经元接种于培养板中，培养至第9天，采用随机数字表法，将其分为7组（ n＝12）：对照组（C组）、脂肪乳剂组（I组）、异丙酚0.1μmol/L组（P1组）、异丙酚1μmol/L组（P2组）、异丙酚10μmol/L组（P3组）、异丙酚100μmol/L组（P4组）和异丙酚1000μmol/L组（P5组）。C组不做任何处理；I组培养液中加入10％脂肪乳剂，终浓度为100μmol/L；P1组、P2组、P3组、P4组、P5组培养液中加入异丙酚，终浓度分别为0.1、1、10、100、1000μmol/L ，继续孵育3 h。于异丙酚孵育前和孵育结束后10 min内采用激光共聚焦显微镜测定[Ca2＋]i ，观察神经元形态和轴突、树突的结构，于异丙酚孵育结束7 d采用Western blot法检测NF-κB的蛋白表达，反映其活性。结果与异丙酚孵育前比较，P2组、P3组、P4组异丙酚孵育结束后各时点[Ca2＋]i升高，P5组异丙酚孵育结束后各时点[Ca2＋]i降低（ P＜0.05），C组、I组和P1组异丙酚孵育结束后各时点[Ca2＋]i差异无统计学意义（ P＞0.05）。与C组比较，P1组、P2组、P3组、P4组、P5组海马神经元NF-κB活性降低（ P＜0.05），I组海马神经元NF-κB 活性差异无统计学意义（ P＞0.05）。C组、I组和P1组海马神经元形态结构正常；P2组、P3组、P4组海马神经元轴突和树突分枝明显减少， P5组海马神经元形态模糊、细胞膜破裂，未见明显的轴突和树突结构。结论异丙酚可通过改变[Ca2＋]i和抑制NF-κB激活，对胎鼠海马神经元产生毒性。Objective To evaluate the effects of propofol on the intracellular calcium ion concentration （[Ca2＋ ]i） and nuclear factor kappa B （NF-κB） activity in hippocampal neurons of fetal rats in vitro .Methods Ten pregnant Sprague-Dawley rats at 16-18 days of gestation ,were sacrificed and the fetal rats were taken out from the abdominal cavity .The hippocampal neurons of the fetal rats were isolated and seeded in culture plates .After being cultured for 9 days ,the neurons were divided into 7 groups （ n=12 each ） using a random number table ：control group （C group） ,intralipid group （I group） and propofol 0.1 ,1 ,10 ,100 ,1 000 μmol/L groups （P1-5 groups） .In group I ,10% intralipid was added to the culture media until the final concentration reached 100μmol/L .In P1-5 groups ,propofol was added to the culture media until the final concentration reached 0.1 ,1 ,10 , 100 and 1 000μmol/L ,respectively .The cells were then incubated for 3 h .The [Ca2＋ ]i and cellular morphology of hippocampal neurons were examined by laser scanning confocal microscopy before incubation with propofol and within 10 min after the end of incubation with propofol .The expression of NF-κB protein in the nucleus was＆nbsp;detected at 7 days after the end of incubation with propofol by Western blot analysis to reflect NF-κB activity . Results Propofol increased [Ca2＋ ]i in P2-4 groups ,while decreased [Ca2＋ ]i in group P5 （ P0.05 ） .The structure of hippocampal neurons was normal in C ,I and P1 groups .The branchings of axons and dendrites in hippocampal neurons were significantly decreased in P 2-4 groups , while the structure of hippocampal neurons became fuzzy , the cell membrane was destroyed and the axons and dendrites were not seen in group P5 .Conclusion Propofol can produce neurotoxic effects on hippocampal neurons of fetal rats by changing the [Ca2＋ ]i and promoting NF-κB activation in vitro .

关 键 词：二异丙酚 胎儿 海马 神经元 钙 NF-ΚB 

分 类 号：R614[医药卫生—麻醉学]