七氟烷下调LSD1表达抑制胚胎干细胞的自我更新

Sevoflurane inhibits embryonic stem cell self-renewal via downregulating expression of lysine-specific demethylase 1

目的:研究七氟烷通过下调赖氨酸去甲基化酶1(lysine-specific demethylase 1,LSD1)的表达影响小鼠胚胎干细胞的自我更新,导致细胞干性下调。方法:将E14小鼠胚胎干细胞随机分为4组(n=6):空白对照组、对照组+LSD1过表达组、七氟烷组和七氟烷+LSD1过表达组。前两组给予含有5%CO_2和21%O_2的混合气体,后两组给予4.1%七氟烷,均处理6 h。实时定量PCR及Western印迹法检测LSD1的表达水平,实时定量PCR检测干性基因Oct4、Sox2、Nanog的表达水平。结果 :与对照组相比,七氟烷组在干预后小鼠胚胎干细胞出现提前分化现象,LSD1及干性基因(Oct4、Sox2、Nanog)表达显著下降(P<0.05)。LSD1过表达逆转七氟烷造成的小鼠胚胎干细胞LSD1及干性基因表达下调。结论:七氟烷可能通过下调小鼠胚胎干细胞LSD1的表达,抑制胚胎干细胞的自我更新,从而影响小鼠胚胎干细胞的正常增殖与分化。

Objective To investigate whether sevoflurane inhibits mouse embryonic stem cells self-renewal and decreases cell stemness via downregulating the expression of lysine-specific demethylase 1（LSD1）. Methods The mouse embryonic stem cells E14 were randomized into four groups（n=6）. The mouse embryonic stem cells in both control group and control＋pre-LSD1 group were treated with 5%CO_2 and 21%O_2 for 6 h. 4.1% sevoflurane was given to the mouse embryonic stem cells for 6 h in both sevoflurane group and sevoflurane ＋pre-LSD1 group. The cells in both pre-LSD1 groups had LSD1 overexpression. Real time PCR and Western blot assay were used to detect the expression of LSD1. The expression level of stemness markers Oct4, Sox2, Nanog was also detected by real time PCR. Results The mouse embryonic stem cells were differentiated in advance and the expression level of LSD1 and stemness markers（Oct4, Sox2, Nanog） reduced significantly（P〈0.05） in sevoflurane group after intervention of sevoflurane when compared with the cells in control group.Overexpression of LSD1 reversed the sevoflurane-induced downregulation of LSD1 and the expression of stemness markers.Conclusions Sevoflurane might inhibit mouse embryonic stem cells self-renewal and differentiation via downregulating the expression of LSD1.