七氟烷对小鼠胚胎神经干细胞增殖的影响  

Effect of sevoflurane on proliferation of forebrain neural stem cells in mice

作　　者：夏贵山 宋琼 宋建强 王朋飞 李保林 XIA Guishan;SONG Qiong;SONG Jianqiang;WANG Pengfei;LI Baolin(Department of Anesthesiology,Zhengzhou Central Hospital,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学附属郑州中心医院郑州市中心医院麻醉科

出　　处：《中华实用诊断与治疗杂志》2019年第12期1168-1172,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省医学科技攻关计划省部共建项目(201701035)

摘　　要：目的探讨七氟烷对小鼠胚胎神经干细胞(forebrain neural stem cells,FNSCs)增殖的影响及机制。方法妊娠第14天小鼠胚胎,分离FNSCs并培养,将对数生长期FNSCs分为对照组、1.2%七氟烷组、2.4%七氟烷组和4.8%七氟烷组,其中对照组吸入体积分数21%O 2、体积分数5%CO 2和平衡N 2,1.2%七氟烷组、2.4%七氟烷组和4.8%七氟烷组在对照组基础上分别吸入体积分数1.2%、2.4%、4.8%七氟烷,吸入七氟烷6h后采用CCK-8法检测各组细胞增殖率,采用BrdU插入实验检测BrdU阳性细胞数,采用流式细胞术检测细胞周期情况,采用Western blot法检测GABA-CREB信号通路GABA、pCREB、CREB蛋白相对表达量。结果吸入七氟烷6h后,4.8%七氟烷组细胞增殖率[(37.0±0.6)%]、BrdU阳性细胞率[(17.23±3.74)%]、GABA和pCREB蛋白相对表达量(31.29±3.22、20.71±2.59)明显低于对照组[(97.0±2.1)%、(22.51±2.53)%、58.26±1.75、29.94±2.45]、1.2%七氟烷组[(86.0±1.7)%、(23.02±3.69)%、59.44±4.29、28.43±3.77]和2.4%七氟烷组[(84.0±1.0)%、(23.15±3.66)%、57.28±3.15、31.74±2.69](P<0.05),G 1/G 0期细胞比率[(78.2±3.7)%]明显高于对照组[(62.3±4.2)%]、1.2%七氟烷组[(63.2±4.2)%]和2.4%七氟烷组[(60.7±1.6)%](P<0.05),CREB蛋白相对表达量(43.78±1.68)与对照组(44.76±2.64)、1.2%七氟烷组(45.48±2.70)、2.4%七氟烷组(44.72±3.41)比较差异无统计学意义(P>0.05);对照组、1.2%七氟烷组、2.4%七氟烷组以上指标组间两两比较差异均无统计学意义(P>0.05)。结论高浓度(4.8%)七氟烷对FNSCs增殖有抑制作用,其机制可能是高浓度七氟烷抑制GABA信号传导,降低CREB磷酸化水平。Objective To investigate the effect of sevoflurane on the proliferation of forebrain neural stem cells(FNSCs)in mice and its possible mechanism.Methods On the 14th day of mice embryonic pregnancy,FNSCs were isolated and cultured.The FNSCs in logarithmic growth phase were divided into control group,1.2%sevoflurane group,2.4%sevoflurane group and 4.8%sevoflurane group.Control group inhaled 21%O 2,5%CO 2 and equilibrium N 2,while 1.2%,2.4%and 4.8%sevoflurane groups inhaled 1.2%,2.4%and 4.8%sevoflurane,respectively,on the basis of the treatment in control group.After inhalation of sevoflurane for 6h,the proliferation rate of each group was detected by CCK-8method.The number of BrdU positive cells was detected by BrdU insertion assay.The cell cycle was detected by flow cytometry.The relative expression levels of GABA,pCREB and CREB proteins in the signaling pathway were detected by Western blot.Results After inhalation of sevoflurane for 6h,the cell proliferation rate((37.0±0.6)%),the BrdU positive rate((17.23±3.74%)),and the relative expressions of GABA and pCREB proteins(31.29±3.22,20.71±2.59)in 4.8%sevoflurane group were significantly lower than those in control group((97.0±2.1)%,(22.51±2.53)%,58.26±1.75,29.94±2.45),1.2%sevoflurane group((86.0±1.7)%,(23.02±3.69)%,59.44±4.29,28.43±3.77)and 2.4%sevoflurane group((84.0±1.0)%,(23.15±3.66)%,57.28±3.15,31.74±2.69)(P<0.05).The proportion of cells in G 1/G 0 phase((78.2±3.7)%)was significantly higher than that in control group((62.3±4.2)%),1.2%sevoflurane group((63.2±4.2)%)and 2.4%sevoflurane group((60.7±1.6)%)(P<0.05),the relative expression of CREB protein showed no significant difference in 4.8%sevoflurane group(43.78±1.68),control group(44.76±2.64),1.2%sevoflurane group(45.48±2.70)and 2.4%sevoflurane group(44.72±3.41)(P>0.05),and all above indexes showed no differences in control group,1.2%sevoflurane group and 2.4%sevoflurane group in multiple comparison(P>0.05).Conclusion High concentration of sevoflurane(4.8%)can inhibit the proliferation of

关 键 词：胚胎神经干细胞 增殖 七氟烷 GABA-CREB信号通路 小鼠 

分 类 号：R614[医药卫生—麻醉学]