七氟烷预处理对大鼠骨髓间充质干细胞在缺氧复氧环境下的作用和机制初探

目的:研究七氟烷预处理对大鼠骨髓间充质干细胞（MSCs）在缺氧复氧环境下的作用和机制初探。方法:分离培养SD大鼠骨髓间充质干细胞，通过流式细胞仪和诱导分化培养鉴定其免疫表型和分化潜能。细胞随机分为3组:组1空白对照组；组2缺氧复氧组（缺氧24 h，复氧24 h）；组3七氟烷预处理组（2%七氟烷预处理2 h后，缺氧24 h，复氧24 h）。处理后收集各组细胞，用流式细胞仪测定其凋亡率，线粒体膜电位变化及细胞周期。Western blot测定细胞外调节蛋白激酶（ERK）的蛋白表达。结果:与组2相比，组3 MSCs凋亡率明显下降，处于S期细胞增多，细胞增殖能力增强。Western blot结果显示:与组2相比，组3磷酸化ERK蛋白表达上升。结论:七氟烷预处理能提高缺氧复氧环境下骨髓间充质干细胞存活率，降低MSCs凋亡数量，并能增强MSCs的增殖能力，这一机制与七氟烷促进MSCs细胞p-ERK蛋白表达相关。

The effects and mechanism involved in sevoflurane pretreatment on rat bone marrow mesenchymal stem cells under hypoxia-reoxygenation environment

Objective:To investigate the effects and mechanism involved in sevoflurane pretreatment on rat bone marrow mesenchymal stem cells (MSCs) under hypoxia-reoxygenation environment.Methods:MSCs were isolated from the SD rat compact bone and then cultured.The immunophenotype and differentiation potential of MSCs were identified by flow cytometry and inductive culture.MSCs were randomly divided into three groups:Control group 1 (cells were cultured in a normoxia),the hypoxia-reoxygenation group 2 (the MSCs were exposed to 24 h hypoxia,followed by 24 h reoxygenation),sevoflurane preconditioning group 3 (cells were exposed to 2 h of 2% sevoflurane,followed by 24 h hypoxia-reoxygenation).MSCs were collected after reoxygenation.The apoptosis,cell cycle and JC-1 was determined by flow cytometry.p-ERK was detected by Western blot.Results:Compared with group 2,the apoptosis rate of group 3 cells decreased significantly.The RNA content of the S-phase was significantly increased and the ability of cell proliferation was enhanced.Western blot results showed that the expression of p-ERK protein in group 3 was higher than that in group 2.Conclusion:Sevoflurane pretreatment can increase the survival rate of bone marrow mesenchymal stem cells under hypoxia and reoxygenation,reduce the apoptosis rate,and enhance the proliferation of MSCs.This mechanism is related to the promotion of p-ERK protein expression in MSCs cells by sevoflurane.