CAJ | 학술논문

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。
목적：평개이병분대체외배양적태서해마신경원내개리자농도（[Ca2＋]i ）화NF-κB활성적영향。방법잉16～18 d SD대서랍경처사，부복취태서，분리해마신경원。장신경원접충우배양판중，배양지제9천，채용수궤수자표법，장기분위7조（ n＝12）：대조조（C조）、지방유제조（I조）、이병분0.1μmol/L조（P1조）、이병분1μmol/L조（P2조）、이병분10μmol/L조（P3조）、이병분100μmol/L조（P4조）화이병분1000μmol/L조（P5조）。C조불주임하처리；I조배양액중가입10％지방유제，종농도위100μmol/L；P1조、P2조、P3조、P4조、P5조배양액중가입이병분，종농도분별위0.1、1、10、100、1000μmol/L ，계속부육3 h。우이병분부육전화부육결속후10 min내채용격광공취초현미경측정[Ca2＋]i ，관찰신경원형태화축돌、수돌적결구，우이병분부육결속7 d채용Western blot법검측NF-κB적단백표체，반영기활성。결과여이병분부육전비교，P2조、P3조、P4조이병분부육결속후각시점[Ca2＋]i승고，P5조이병분부육결속후각시점[Ca2＋]i강저（ P＜0.05），C조、I조화P1조이병분부육결속후각시점[Ca2＋]i차이무통계학의의（ P＞0.05）。여C조비교，P1조、P2조、P3조、P4조、P5조해마신경원NF-κB활성강저（ P＜0.05），I조해마신경원NF-κB 활성차이무통계학의의（ P＞0.05）。C조、I조화P1조해마신경원형태결구정상；P2조、P3조、P4조해마신경원축돌화수돌분지명현감소， P5조해마신경원형태모호、세포막파렬，미견명현적축돌화수돌결구。결론이병분가통과개변[Ca2＋]i화억제NF-κB격활，대태서해마신경원산생독성。
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 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 ( P<0.05 ) .Compared with group C ,the activity of NF-κB was significantly decreased in P1-5 groups ( P<0.05 ) ,while no significant change was found in C and I groups ( P>0.05 ) .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 .