目的·探索多次七氟烷处理对原代少突胶质细胞增殖和分化的影响。方法·提取出生当日大鼠皮层的少突胶质前体细胞（oligodendrocyte precursor cell，OPC）并进行体外培养。细胞分为对照组和七氟烷处理组。为了模拟临床使用七氟烷的情况，将七氟烷组细胞使用3%七氟烷连续处理3 d，每日1次，每次处理2 h。OPC分化成熟为少突胶质细胞后，使用免疫荧光染色和蛋白质印迹法（Western blotting）检测髓鞘碱性蛋白（myelin basic protein，MBP）和髓鞘关联糖蛋白（myelin-associated glycoprotein，MAG）的表达情况。采用细胞增殖实验（BrdU、Ki67染色）、细胞存活率实验（CCK8）检测七氟烷对OPC增殖能力和少突胶质细胞存活率的影响。采用Western blotting检测半胱氨酸蛋白酶-3（caspase-3）的蛋白含量。使用慢病毒转染技术，在OPC内过表达YTH N⁶-甲基腺苷RNA结合蛋白1（YTH N⁶-methyladenosine RNA binding protein F1，YTHDF1），后采用CCK8检测细胞增殖和存活情况。结果·免疫荧光结果提示，反复暴露于七氟烷会导致表达成熟髓鞘表面标志物（MBP、MAG）的原代少突胶质细胞数量减少；Western blotting结果表明，多次七氟烷处理导致原代OPC中caspase-3表达上调；CCK8结果表明，随着七氟烷处理次数的增加，原代OPC的存活率下降；然而，BrdU、Ki67染色结果显示，原代OPC在七氟烷处理后增殖能力增强。此外，过表达YTHDF1可以部分改善多次七氟烷处理而导致的原代OPC存活率下降（均P<0.05）。结论·多次七氟烷处理损伤原代少突胶质细胞的成髓鞘能力和存活率，表现为部分原代OPC凋亡；同时七氟烷处理代偿性提高了存活的原代OPC的增殖能力。

Objective ·To investigate the effects of multiple sevoflurane exposures on the proliferation and differentiation of primary oligodendrocytes. Methods ·Oligodendrocyte precursor cells (OPCs) were extracted from the cortex of rats on the day of birth and cultured in vitro. The cells were divided into control and sevoflurane groups. To simulate the clinical situation of sevoflurane exposure, cells in the sevoflurane group were exposed to 3% sevoflurane for 3 consecutive days, 2 h for each time. After the OPCs were differentiated and matured, immunofluorescence staining and Western blotting were used to detect the expression of myelin basic protein (MBP) and the myelin-associated glycoprotein (MAG). Cell proliferation assays (BrdU and Ki67) and a cell viability assay (CCK8) were used to detect the effects of sevoflurane on the proliferation ability of OPCs and the survival rate of oligodendrocytes. Western blotting was used to detect the protein content of caspase-3. Lentiviral transfection technology was used to overexpress YTH N⁶-methyladenosine RNA binding protein F1 (YTHDF1) in OPCs, and then CCK8 was used to detect cell proliferation and survival. Results ·Immunofluorescence results showed that multiple sevoflurane exposures led to a decrease in the number of primary oligodendrocytes expressing mature myelin surface markers (MBP, MAG); Western blotting results showed that these exposures led to upregulation of caspase-3 expression in primary OPCs; CCK8 results showed that the survival rate of primary OPCs decreased with the increase in the number of sevoflurane treatments; however, BrdU and Ki67 staining results showed that the proliferation ability of primary OPCs was enhanced after sevoflurane exposure. In addition, overexpression of YTHDF1 could partially improve the decreased survival rate of primary OPCs caused by multiple sevoflurane exposures (all P<0.05). Conclusion ·Multiple sevoflurane exposures impair the myelinating ability and survival rate of primary oligodendrocytes, manifested by apoptosis of some primary OPCs. In contrast, sevoflurane exposure compensatorily increases the proliferation ability of surviving primary OPCs.