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作 者:钟玉玲[1] 梁羽冰[1] 利莉[1] 陈静[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2期140-142,共3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 评价异丙酚对胎鼠离体海马神经元凋亡的影响.方法 体外培养胎鼠海马神经元,调整密度为5×104个/ml后接种到96孔板和放有圆形玻片的24孔板中,加入培养液培育,采用随机数字表法分为5组(n=18):对照组(C组)、脂肪乳剂组(Ⅰ组)、异丙酚1组(P1组)、异丙酚2组(P2组)和异丙酚3组(P3组).C组不做任何处理,Ⅰ组在培养液中加入10%脂肪乳剂,终浓度为100μmol/L,P1组、P2组和P3组在培养液中加入异丙酚,终浓度分别为1、10和100 μmol/L,继续孵育3h.采用流式细胞术检测细胞凋亡情况,采用RT-PCR检测凋亡相关因子Bcl-2 mRNA和caspase-3 mRNA的表达,Western blot法检测Bcl-2蛋白和actived-easpase-3蛋白的表达.结果 与C组比较,P1组、P2组和P3组海马神经元凋亡率升高,Bcl-2 mRNA和蛋白表达下调,caspase-3 mRNA和actived-caspase-3蛋白表达上调(P<0.05),Ⅰ组上述各指标差异无统计学意义(P>0.05).结论 异丙酚通过抑制Bcl-2表达,增强caspase-3活性促进胎鼠离体海马神经元凋亡.Objective To evaluate the effects of propofol on apoptosis in the hippocampal neurons of fetal rats in vitro.Methods The isolated hippocampal neurons were seeded into 96-well plates or 24-well plates at a density of 5 × 104 cells/ml.The cells were randomly divided into 5 groups (n =18 each) using a random number table:control group (group C),in tralipid group (group Ⅰ) and propofol 1,10,100 μmol/L groups (P1-3 groups).In group Ⅰ,10% intralipid was added to the culture media until the final concentration reached 100 μmol/L.In groups P1-3,propofol was added to the culture media until the final concentration reached 1,10 and 100 μmol/L,respectively,and the cells were then incubated for 3 h.The cell apoptosis was assessed by flow cytometry.The expression of Bcl-2 mRNA and caspase-3 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).The expression of Bcl-2 and actived-caspase-3 protein was determined by Western blot analysis.The apoptosis rate was calculated.Results Compared with group C,the apoptosis rate was significantly increased,the expression of Bcl-2 mRNA and protein was down-regulated,and the expression of caspase-3 mRNA and actived-caspase-3 protein was up-regulated in P1-3 groups (P 〈 0.05).There was no significant difference in the parameters mentioned above between group Ⅰ and group C (P 〉 0.05).Conclusion Propofol induces apoptosis in isolated hippocampal neurons by inhibiting Bcl-2 expression and enhancing caspase-3 activity in fetal rats.
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