miR-214对异丙酚麻醉大鼠术后神经元损伤的保护作用  

Protective effect of miR-214 on neuronal injury in propofol-anesthetized thoracotomy rats

作　　者：杨继梅 卢文江 周娇洁 YANG Jimei;LU Wenjiang;ZHOU Jiaojie(Department of Anesthesiology,Leshan People’s Hospital,Leshan 614000,China)

机构地区：[1]乐山市人民医院麻醉科,四川乐山614000

出　　处：《中国比较医学杂志》2022年第9期39-46,81,共9页Chinese Journal of Comparative Medicine

基　　金：四川省卫生适宜技术推广项目(19SYJS56)。

摘　　要：目的 探讨miR-214对异丙酚麻醉诱导的神经元损伤的保护作用及生物学机制。方法 7日龄SD雄性大鼠随机分为4组(每组15只):生理盐水组(NS)、异丙酚麻醉开腹探查组(模型)、miR-NC组和miR-214组。除NS组外,其余组别建立异丙酚麻醉开腹探查模型。miR-NC组和miR-214组分别在麻醉前将miR-NC-agomir或miR-214-agomir注射到海马内。采用免疫荧光法检测海马mTORC1的表达,TUNEL染色检测海马组织细胞凋亡,RT-qPCR分析海马组织中miR-214表达。分别将miR-214抑制剂和mTORC1抑制剂转染入HT22海马神经元细胞,然后暴露于异丙酚。采用流式细胞术分析细胞的存活情况和Western blot分析TEFB、C-caspase3蛋白表达。结果 与NS组相比,模型组大鼠海马中miR-214明显下调(P<0.05),并且海马神经细胞凋亡显著增加(P<0.05)。与模型组相比,miR-214组海马神经元凋亡减弱(P<0.05)。生物信息学预测证明mTORC1和miR-214之间存在特异性结合位点。免疫荧光检测显示,与NS组比较,模型组诱导海马神经组织中mTORC1表达增加(P<0.05),miR-214治疗显著降低了mTORC1表达(P<0.05)。此外,与NC对照组相比,si-mTORC1转染导致暴露于异丙酚的HT22海马神经元细胞的凋亡率显著降低,并且TFEB表达显著增加(P<0.01),以及C-caspase3降低(P<0.05)。而miR-214抑制剂转染显著逆转了si-mTORC1的保护作用以及诱导的TFEB、C-caspase3蛋白表达的变化(P<0.05)。结论 miR-214可能通过mTORC1-TFEB通路减轻异丙酚神经毒性,提高神经元的存活率。Objective To investigate the protective effect and biological mechanism of miR-214 on neuronal injury induced by propofol anesthesia,and elucidate the underlying mechanisms of this process.Methods Seven-day-old male Sprague-Dawley rats were randomly divided into four groups(15 rats per group):normal saline(NS),propofol anesthesia thoracotomy exploration(model),miR-NC and miR-214.With the exception of the NS group,all groups underwent establishment of the propofol anesthesia thoracotomy exploration model.In miR-NC and miR-214 groups,miR-NC-agomir or miR-214-agomir,respectively,were injected into hippocampus before anesthesia.Expression of mTORC1 in the hippocampus was detected by immunofluorescence,apoptosis was detected by TUNEL staining,and expression of miR-214 in hippocampus was analyzed by RT-qPCR.HT22 hippocampal neurons were transfected with an miR-214 inhibitor and mTORC1 inhibitor,and then exposed to propofol.Flow cytometry was used to analyze cell survival,whereas Western blot was used to analyze protein expression of TEFB and C-caspase3.Results Compared with the NS group,miR-214 in the hippocampus of the model group was significantly downregulated(P<0.05) and apoptosis of hippocampal neurons was significantly increased(P<0.05).Compared with the model group,apoptosis of hippocampal neurons in the miR-214 group was decreased(P<0.05).Bioinformatics prediction indicated the presence of a specific binding site between mTORC1 and miR-214.Compared with the NS group,expression of mTORC1 was increased in the model group(P<0.05),and miR-214 treatment significantly reduced expression of mTORC1(P<0.05).In addition,compared with the NC group,si-mTORC1 transfection significantly reduced the apoptosis rate of HT22 hippocampal neurons exposed to propofol(P<0.05),increased TFEB expression(P<0.01),and decreased cleaved caspase 3(P<0.05).miR-214 inhibitor transfection significantly reversed the protective effect of si-mTORC1 and changes of TFEB and C-caspase3 protein expression induced by si-mTORC1(P<0.05).Conclusions miR

关 键 词：miR-214 异丙酚 开腹探查 大鼠 神经元 mTORC1-TFEB通路 

分 类 号：R-33[医药卫生]