七氟烷对胶质瘤U87细胞侵袭能力的影响及机制研究

Effect of Sevoflurane on Invasion Ability of Glioma U87 Cells and Its Mechanism

目的探讨七氟烷对U87细胞侵袭能力的影响及机制。方法于2022年9月—2023年6月,采用徐州医科大学麻醉学实验室提供的U87细胞培养至对数生长期。Transwell实验取对数生长期细胞分成8份,然后随机分为对照(Con)组和七氟烷(Sev)组,每组4份进行平行实验。培养的细胞置于麻醉箱内,七氟烷组通载体气体和1.4%七氟烷4 h,对照组只通载体气体,观察七氟烷对U87细胞侵袭能力的影响;免疫印迹实验取对数生长期细胞分成12份,然后随机分为对照组,七氟烷2 h组和七氟烷4 h组,每组4份进行平行实验,观察七氟烷对pERK1/2、ERK1/2表达水平的影响;细胞免疫荧光化学实验取对数生长期细胞分成8份,然后随机分为对照组和七氟烷组(4 h),每组4份进行平行实验,检测七氟烷对pERK1/2、Nrf2在细胞分布的影响。结果与对照组相比,七氟烷组U87细胞穿过微孔膜的数目减少[Con组(100.000±0.000)%,Sev组(34.180±3.342)%],差异有统计学意义(t=16.697,P<0.05)。与对照组相比,七氟烷组pERK1/2的灰度值下降[Con组(100.000±0.000)%,Sev2 h组(0.196±0.028)%,Sev4 h组(0.073±0.025)%],差异有统计学意义(F=546.886,P<0.05),且核内pERK1/2[Con组(21.903±1.776)%,Sev组(8.412±0.367)%]及核内Nrf2表达水平均显著下降[Con组(18.083±0.788)%,Sev组(12.315±1.604)%],差异有统计学意义(t=7.438、3.228,P均<0.05)。结论七氟烷通过抑制ERK1/2-Nrf2信号通路抑制人胶质细胞瘤U87侵袭能力。

Objective To explore the impact of sevoflurane on U87 cells migration.Methods U87 cells,which pro-vided by anesthesiology Laboratory of Xuzhou Medical University were cultured to logarithmic growth stage and then randomly random grouping for all experiments.In Transwell experiment,cells were divided into 8 parts,and then ran-domly divided into control(Con)and sevoflurane(Sev)groups(with 4 parts in each group for parallel experiments)to detect the impact of sevoflurane on the migration between September 2022 and June 2023.Cultured cells were placed in an anaesthetic chamber 1.4%sevoflurane with carrier gas in the sevoflurane group and carrier gas only in the con-trol group for 4 h.The Transwel assay was used to evaluate the migration of U87 cells;In WB experiment,cells were divided into 12 parts,and then randomly divided into control group,sevoflurane 2 h group and sevoflurane 4 h group(with 4 parts in each group for parallel experiments)to evaluate the level of pERK1/2 and ERK1/2 expression;In im-munofluorescence experiment,cells at logarithmic growth stage were divided into 8 parts,and then randomly divided into control and sevoflurane groups(4 h)(with 4 parts in each group for parallel experiments)to detect pERK1/2 and Nrf2 cell distribution.Results Compared with the control group,the number of U87 cells passing through the micropo-rous membrane in sevoflurane group decreased[Con group(100.000±0.000)%,Sev group(34.180±3.342)%],and the difference was statistically significant(t=16.697,P<0.05).Compared with the control group,the gray value of pERK1/2 in sevoflurane group decreased[Con group(100.000±0.000)%,Sev2 h group(0.196±0.028)%,Sev4 h group(0.073±0.025)%],and the difference was statistically significant(F=546.886,P<0.05).The expression levels of pERK1/2[Con group(21.903±1.776)%,Sev group(8.412±0.367)%]and Nrf2 were significantly decreased[Con group(18.083±0.788)%,Sev group(12.315±1.604)%],the differences were statistically significant(t=7.438,3.228,both P<0.05).Conclusion Sevoflurane inhibits the invasion of human glioma U87 by inhibiting ERK1/2-Nrf2 signal pathway.