CAJ | 학술논문

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.
목적 평개이병분대태서리체해마신경원조망적영향.방법 체외배양태서해마신경원,조정밀도위5×104개/ml후접충도96공판화방유원형파편적24공판중,가입배양액배육,채용수궤수자표법분위5조(n=18):대조조(C조)、지방유제조(Ⅰ조)、이병분1조(P1조)、이병분2조(P2조)화이병분3조(P3조).C조불주임하처리,Ⅰ조재배양액중가입10％지방유제,종농도위100μmol/L,P1조、P2조화P3조재배양액중가입이병분,종농도분별위1、10화100 μmol/L,계속부육3h.채용류식세포술검측세포조망정황,채용RT-PCR검측조망상관인자Bcl-2 mRNA화caspase-3 mRNA적표체,Western blot법검측Bcl-2단백화actived-easpase-3단백적표체.결과 여C조비교,P1조、P2조화P3조해마신경원조망솔승고,Bcl-2 mRNA화단백표체하조,caspase-3 mRNA화actived-caspase-3단백표체상조(P＜0.05),Ⅰ조상술각지표차이무통계학의의(P＞0.05).결론 이병분통과억제Bcl-2표체,증강caspase-3활성촉진태서리체해마신경원조망.
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.