Effect of apoptosis in neural stem cells treated with sevoflurane.
Effect of apoptosis in neural stem cells treated with sevoflurane.
复制标题
七氟烷处理神经干细胞凋亡的影响
DOI：
10.1186/s12871-015-0018-8
复制
发表时间：
2015
影响因子：
2.2
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Qiu J;Shi P;Mao W;Zhao Y;Liu W;Wang Y
背景目前，七氟醚吸入麻醉在婴幼儿中的应用较为广泛。但长期吸入麻醉药可引起神经系统功能紊乱，尤其是婴幼儿和发育中的脑神经退行性变。婴幼儿中枢神经系统的退化可影响记忆和认知功能。γ-氨基丁酸（γ-Aminobutyric acid，GABA）是中枢神经系统的一种抑制性神经递质。吸入麻醉药七氟醚可能激活GABA A受体，抑制中枢神经系统，导致神经细胞凋亡变性，在脑发育的关键时期出现认知功能障碍。将第3代神经干细胞按培养液随机分为4组：七氟醚组（S组）、GABA A受体拮抗剂、荷包牡丹碱组（B组）、七氟醚+GABA A受体拮抗剂、荷包牡丹碱组（S + B组）、二甲基亚砜组（D组）。B组和D组不接受七氟醚预处理。S组和S + B组分别给予1个最低肺泡浓度（MAC）的七氟醚预处理0 h、3 h、6 h和12 h。S + B组和B组用荷包牡丹碱（10 μ M）预处理。D组用DMSO（10 μ L/mL）处理。经上述处理后，各组均培养48 h。采用CCK-8法检测细胞活力，乳酸脱氢酶（LDH）法检测细胞毒性，流式细胞术检测细胞凋亡率，免疫印迹法检测GABAAR、抗凋亡蛋白Bcl-2、促凋亡蛋白Bax和Caspase-3的表达。1 MAC作用12 h后，细胞活力明显下降，细胞毒作用呈时间依赖性增加。Annexin V/PI双染流式细胞仪检测细胞凋亡率增加。GABAA受体、促凋亡蛋白Bax和凋亡蛋白Caspase-3蛋白水平升高，抗凋亡蛋白Bcl-2蛋白水平降低。结论七氟烷可通过激活GABAA抑制中枢神经系统，导致神经干细胞凋亡，从而导致NSCs变性。
BackgroundAt present, sevoflurane inhalation anesthesia used on infants is well-known. But long-time exposure to inhalation anesthetic could cause neurologic disorder, especially nerve degeneration in infant and developing brain. The central nervous system degeneration of infants could affect the memory and cognitive function. γ-Aminobutyric acid (GABA) is a known inhibitory neurotransmitter in central nervous system. Inhalation anesthetic sevoflurane may activate GABAAreceptor to inhibit central nervous system, leading to apoptosis of neural degeneration, cognitive dysfunction in the critical period of brain development.MethodsNeural stem cells were derived from Wistar embryos, culturedin vitro. Third generation of neural stem cells were randomly divided into four groups according to cultured suspension: Sevoflurane group (Group S), GABAAreceptor antagonists, Bicuculline group (Group B), Sevoflurane + GABAAreceptor antagonists, Bicuculline group (Group S + B), dimethyl sulphoxide (DMSO) group (Group D). Group B and Group D did not receive sevoflurane preconditioning. Group S and Group S + B were pretreated with 1 minimum alveolar concentration (MAC) sevoflurane for 0 h, 3 h, 6 h, and 12 h. Group S + B and Group B were pretreated with bicuculline (10 uM). Group D was treated with DMSO (10 uL/mL). After treatments above, all groups were cultured for 48 h. Then we measured the cells viability by Cell Counting Kit (CCK-8) assay, cytotoxicity by Lactate Dehydrogenase (LDH) assay, apoptosis ratio with Annexin V/propidium iodide (PI) staining by flow cytometry, and the expression of GABAAR, anti-apoptotic protein Bcl-2, pro-apoptotic protein Bax and Caspase-3 by western blotting.ResultsAfter exposing to sevoflurane for 0 h, 3 h, 6 h, and 12 h with 1MAC, we found that cell viability obviously decreased and cytotoxicity increased in time-dependent way. And Annexin V/PI staining indicated increased apoptosis ratio by flow cytometry. The protein level of GABAAreceptor, pro-apoptotic protein Bax and apoptosis protein Caspase-3 increased; while anti-apoptotic protein Bcl-2 decreased. And bicuculline could reverse all detrimental results caused by sevoflurane.ConclusionSevoflurane can inhibit the central nervous system by activating GABAA, resulting in apoptosis of neural stem cells, thus leading to the NSCs degeneration.
登录
查看更多内容
影响因子：
8.8
作者：
Ebert, TJ;Robinson, BJ;Pichotta, PJ
通讯作者：
Pichotta, PJ
七氟醚对培养神经干细胞自我更新能力和分化的影响
影响因子：
4.4
作者：
Nie, Huang;Peng, Zhengwu;Xiong, Lize
通讯作者：
Xiong, Lize
DOI：
10.1053/j.jvca.2012.04.018
发表时间：
2012-10-01
影响因子：
2.8
作者：
Landoni, Giovanni;Rodseth, Reitze N.;Zangrillo, Alberto
通讯作者：
Zangrillo, Alberto
DOI：
10.1053/j.jvca.2009.01.022
发表时间：
2009-10-01
影响因子：
2.8
作者：
Bignami, Elena;Biondi-Zoccai, Giuseppe;Zangrillo, Alberto
通讯作者：
Zangrillo, Alberto
影响因子：
8.8
作者：
Stucke, AG;Zuperku, EJ;Stuth, EAE
通讯作者：
Stuth, EAE