国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2014年
4期
294-299
,共6页
目的 探讨不同浓度异丙酚对胎鼠离体海马神经元存活及凋亡的影响. 方法 SD大鼠妊娠17 d～18 d后取胎鼠海马神经元体外培养7d,应用免疫细胞化学法鉴定神经元并作为研究对象.①采用随机数字表法将其随机分为12组(每组设5个平行副孔,实验重复5次):对照组(C组)正常培养,异丙酚溶剂对照组(D组)用含0.1％二甲基亚砜(dimethyl sulfoxide,DMSO)的培养基处理,不同浓度异丙酚组(P1～10组)分别用含异丙酚终浓度为0.01、0.1、0.5、1、5、10、50、100、150、200μmol/L的培养基换液处理,继续培养24 h后用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]法检测细胞生存率;②采用随机数字表法将其随机分为7组(实验重复5次):对照组(C组)正常培养,异丙酚溶剂对照组(D组)用含0.1％DMSO的培养基处理,不同浓度异丙酚组(PⅠ、Ⅱ、Ⅲ、Ⅳ、Ⅴ组)分别用含异丙酚终浓度为0.1、l、10、100、200μmol/L的培养基换液处理,继续培养24 h,免疫荧光细胞化学染色观察细胞核形态学结果,Annexin-V-FLUOS双染流式细胞术检测海马神经元凋亡率. 结果 ①与C组比较,D组海马神经元生存率差异无统计学意义(P＞0.05);与D组比较,P1、2、5组生存率差异无统计学意义(P＞0.05),P3组(102.2±2.7)％、P4组(102.9±2.7)％生存率升高(P＜0.05),而P6～10组生存率分别为(97.0±3.5)％、(90.1±1.7)％、(79.8±0.9)％、(67.9±1.1)％、(42.3±2.2)％,呈浓度依赖性降低(P＜0.05或P＜0.01);②与C组比较,D组海马神经元凋亡率差异无统计学意义(P＞0.05);与D组比较,PⅠ组凋亡率差异无统计学意义(P＞0.05),PⅡ组(4.7±0.7)％凋亡率明显降低(P＜0.05),而PⅢ、Ⅳ、Ⅴ组凋亡率分别为(8.2±0.8)％、(18.1±1.1)％、(29.1±2.1)％,呈浓度依赖性升高(P＜0.05或P＜0.01). 结论 异丙酚对胎鼠离体海马神经元生存及凋亡的影响呈双向作用,1 μmol/L浓度异丙酚能够对抗海马神经元凋亡,提高细胞存活率,而10、100、200 μmol/L浓度的异丙酚则呈剂量依赖性地促进海马神经元凋亡,降低细胞生存率.
目的 探討不同濃度異丙酚對胎鼠離體海馬神經元存活及凋亡的影響. 方法 SD大鼠妊娠17 d～18 d後取胎鼠海馬神經元體外培養7d,應用免疫細胞化學法鑒定神經元併作為研究對象.①採用隨機數字錶法將其隨機分為12組(每組設5箇平行副孔,實驗重複5次):對照組(C組)正常培養,異丙酚溶劑對照組(D組)用含0.1％二甲基亞砜(dimethyl sulfoxide,DMSO)的培養基處理,不同濃度異丙酚組(P1～10組)分彆用含異丙酚終濃度為0.01、0.1、0.5、1、5、10、50、100、150、200μmol/L的培養基換液處理,繼續培養24 h後用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴鹽[3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]法檢測細胞生存率;②採用隨機數字錶法將其隨機分為7組(實驗重複5次):對照組(C組)正常培養,異丙酚溶劑對照組(D組)用含0.1％DMSO的培養基處理,不同濃度異丙酚組(PⅠ、Ⅱ、Ⅲ、Ⅳ、Ⅴ組)分彆用含異丙酚終濃度為0.1、l、10、100、200μmol/L的培養基換液處理,繼續培養24 h,免疫熒光細胞化學染色觀察細胞覈形態學結果,Annexin-V-FLUOS雙染流式細胞術檢測海馬神經元凋亡率. 結果 ①與C組比較,D組海馬神經元生存率差異無統計學意義(P＞0.05);與D組比較,P1、2、5組生存率差異無統計學意義(P＞0.05),P3組(102.2±2.7)％、P4組(102.9±2.7)％生存率升高(P＜0.05),而P6～10組生存率分彆為(97.0±3.5)％、(90.1±1.7)％、(79.8±0.9)％、(67.9±1.1)％、(42.3±2.2)％,呈濃度依賴性降低(P＜0.05或P＜0.01);②與C組比較,D組海馬神經元凋亡率差異無統計學意義(P＞0.05);與D組比較,PⅠ組凋亡率差異無統計學意義(P＞0.05),PⅡ組(4.7±0.7)％凋亡率明顯降低(P＜0.05),而PⅢ、Ⅳ、Ⅴ組凋亡率分彆為(8.2±0.8)％、(18.1±1.1)％、(29.1±2.1)％,呈濃度依賴性升高(P＜0.05或P＜0.01). 結論 異丙酚對胎鼠離體海馬神經元生存及凋亡的影響呈雙嚮作用,1 μmol/L濃度異丙酚能夠對抗海馬神經元凋亡,提高細胞存活率,而10、100、200 μmol/L濃度的異丙酚則呈劑量依賴性地促進海馬神經元凋亡,降低細胞生存率.
목적 탐토불동농도이병분대태서리체해마신경원존활급조망적영향. 방법 SD대서임신17 d～18 d후취태서해마신경원체외배양7d,응용면역세포화학법감정신경원병작위연구대상.①채용수궤수자표법장기수궤분위12조(매조설5개평행부공,실험중복5차):대조조(C조)정상배양,이병분용제대조조(D조)용함0.1％이갑기아풍(dimethyl sulfoxide,DMSO)적배양기처리,불동농도이병분조(P1～10조)분별용함이병분종농도위0.01、0.1、0.5、1、5、10、50、100、150、200μmol/L적배양기환액처리,계속배양24 h후용3-(4,5-이갑기새서-2)-2,5-이분기사담서추염[3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]법검측세포생존솔;②채용수궤수자표법장기수궤분위7조(실험중복5차):대조조(C조)정상배양,이병분용제대조조(D조)용함0.1％DMSO적배양기처리,불동농도이병분조(PⅠ、Ⅱ、Ⅲ、Ⅳ、Ⅴ조)분별용함이병분종농도위0.1、l、10、100、200μmol/L적배양기환액처리,계속배양24 h,면역형광세포화학염색관찰세포핵형태학결과,Annexin-V-FLUOS쌍염류식세포술검측해마신경원조망솔. 결과 ①여C조비교,D조해마신경원생존솔차이무통계학의의(P＞0.05);여D조비교,P1、2、5조생존솔차이무통계학의의(P＞0.05),P3조(102.2±2.7)％、P4조(102.9±2.7)％생존솔승고(P＜0.05),이P6～10조생존솔분별위(97.0±3.5)％、(90.1±1.7)％、(79.8±0.9)％、(67.9±1.1)％、(42.3±2.2)％,정농도의뢰성강저(P＜0.05혹P＜0.01);②여C조비교,D조해마신경원조망솔차이무통계학의의(P＞0.05);여D조비교,PⅠ조조망솔차이무통계학의의(P＞0.05),PⅡ조(4.7±0.7)％조망솔명현강저(P＜0.05),이PⅢ、Ⅳ、Ⅴ조조망솔분별위(8.2±0.8)％、(18.1±1.1)％、(29.1±2.1)％,정농도의뢰성승고(P＜0.05혹P＜0.01). 결론 이병분대태서리체해마신경원생존급조망적영향정쌍향작용,1 μmol/L농도이병분능구대항해마신경원조망,제고세포존활솔,이10、100、200 μmol/L농도적이병분칙정제량의뢰성지촉진해마신경원조망,강저세포생존솔.
Objective To investigate the effect of different concentration of propofol on cell viability in hippocampal neurons of fetal rats in vitro.Methods Primary cultured hippocampal neurons were prepared from the fetuses of Sprague-Dawley rats after 17 d-18 d of gestation,and cultured for 7 d,immunocytochemical method was used to identify the cultured neurons.① The neurons were randomly divided into 12 groups (n=5):control group (group C) received no treatment,in group D 0.1％ dimethyl sulfoxide (DMSO) was added,in group P1-10 propofol was added at the final concentration of 0.01,0.1,0.5,1,5,10,50,100,150 μmol/L and 200 μmol/L respectively.The cells were selected after 24 h incubation for detection of the cell survival rate by cell viability[3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay].② The neurons were randomly divided into 7 groups(n=5):control group (group C) received no treatment,in group D 0.1％ DMSO was added,in group PⅠ,Ⅱ,Ⅲ,v,v propofol was added at the final concentration of 0.1,1,10,100 μmol/L and 200 μmol/L respectively.The cells were selected after 24 h incubation for detection of the neuronal apoptosis by immunofluorescence staining and flow cytometry.Results ① There was no significantly difference in the cell survival rate between group C and D (P＞0.05).Compared with group D,the cell survival rate was no significantly difference in group P1,2,5(P＞0.05),the cell survival rate was significantly increased in group P3(102.2±2.7)％,P4(102.9±2.7)％(P＜0.05) and that was significantly lower in group P6-10 in a concentration-dependent manner by (97.0±3.5)％,(90.1 ± 1.7)％,(79.8±0.9)％,(67.9± 1.1)％,(42.3±2.2)％ respectively (P＜0.05 or P＜0.01).② There was no significantly difference in the neuronal apoptosis rate between group C and D (P＞0.05).Compared with group D,the neuronal apoptosis rate was no significantly difference in group PⅠ (P＞0.05),the neuronal apoptosis rate was significantly decreased in group PⅡ (P＜0.05) and that was significantly higher in group PⅢ,Ⅳ,Ⅴ in a concentration-dependent manner by (8.2±0.8)％,(18.1±1.1)％,(29.1±2.1)％ respectively(P＜0.05 or P＜0.01).Conclusions The effect of propofol on cell viability and apoptosis in hippocampal neurons was bidirectional,propofol at the concentration of 1 μmol/L can inhibit the apoptosis and improve the survival rate of neurons,while decrease the cell viability through promoting the cell apoptosis in a concentration-dependent manner at the concentration of 10,100 μmol/L and 200 μmol/L.
