七氟醚联合顺铂激活IP_(3)R-1依赖的人神经母细胞瘤细胞凋亡

Sevoflurane combined with cisplatin activates IP_(3)R-1 dependent neuronal apoptosis

目的探讨吸入七氟醚对低浓度顺铂预处理的人神经母细胞瘤(SH-SY5Y)细胞增殖活性的影响,并讨论相关机制。方法建立低浓度顺铂预处理细胞模型,噻唑蓝比色法检测不同浓度梯度七氟醚对SH-SY5Y细胞增殖活性的影响,利用Western blotting检测Caspase-3和LC3Ⅱ蛋白表达,利用钙成像技术分析细胞内钙离子浓度变化。结果低浓度顺铂预处理后,0.6%七氟醚对SH-SY5Y细胞增殖活性未产生影响,但可诱导细胞LC3-Ⅱ表达增加,而1.7%七氟醚显著抑制SH-SY5Y细胞增殖活性(P<0.01),诱导LC3-Ⅱ和Caspase-3表达增加(P<0.01),并且增加胞内钙离子浓度(P<0.01);当部分干扰IP_(3)R-1表达后,1.7%七氟醚诱发的LC3-Ⅱ和Caspase-3表达减少(P<0.05)。结论1.7%七氟醚诱导低浓度顺铂预处理SH-SY5Y增殖活性的降低,可能与IP_(3)R-1介导胞内钙离子浓度变化致细胞凋亡增加以及自噬水平增强有关。

Objective To investigate the effect of inhaled anesthetic sevoflurane on the apoptosis of cisplatin-pretreated human neuroblastoma(SH-SY5Y)cells,and to analyze the possible mechanism.Methods The low-concentration cisplatin pretreated cell model was established.Cell viability was analyzed by thiazole blue colorimetry.The protein expression levels of Caspase-3 and LC3-Ⅱwere detected by Western blotting.Changes in intracellular Ca^(2+) concentration were analyzed by calcium imaging technology.Results After cisplatin pretreatment,0.6%sevoflurane induced an increase in the expression of LC3-Ⅱin SH-SY5Y,while 1.7%sevoflurane significantly inhibited cell viability(P<0.01),with increased expression of LC3-Ⅱand Caspase-3(P<0.01)and intracellular Ca^(2+)concentration(P<0.01).After partially interfered with IP_(3)R-1,the expression of LC3-Ⅱand Caspase-3 induced by 1.7%sevoflurane decreased significantly(P<0.05).Conclusion 1.7%sevoflurane inhibited proliferation of low concentration cisplatin pretreated SH-SY5Y by triggering IP_(3)R-1 mediated intracellular calcium oscillation and apoptosisi,and promoting autophagy.