右美托咪定通过miR-204-5p/BDNF/TrkB轴抑制七氟醚麻醉诱导HT22细胞损伤的作用机制研究  

Mechanism of Dexmedetomidine Inhibiting Sevoflurane Anesthesia-Induced HT22 Cell Damage via miR-204-5p/BDNF/TrkB Axis

作　　者：张云 康林莉 ZHANG Yun;KANG Linli(Pangang Group General Hospital,Panzhihua,Sichuan,China 617000)

机构地区：[1]攀钢集团总医院,四川攀枝花617000

出　　处：《中国药业》2025年第7期50-58,共9页China Pharmaceuticals

基　　金：2020年四川省医学(青年创新)科研课题项目[S20271]。

摘　　要：目的探讨右美托咪定(Dex)对七氟醚(Sev)诱导的小鼠HT22海马神经元细胞氧化损伤和神经炎症的影响,以及miR-204-5p/脑源性神经营养因子(BDNF)/酪氨酸激酶受体B(TrkB)轴可能的调控机制。方法将HT22细胞用10μmol/L Dex预处理2 h后,用3.4%Sev处理6 h。将miR-204-5p mimic,miR-204-5p inhibitor,BDNF siRNA或相应阴性对照分别转染至HT22细胞,再进行Dex和/或Sev处理。采用CCK-8法检测细胞活力,分别采用乳酸脱氢酶(LDH)细胞毒性试剂盒、丙二醛(MDA)检测试剂盒、白细胞介素6(IL-6)和肿瘤坏死因子-α(TNF-α)酶联免疫吸附试验(ELISA)法检测试剂盒检测神经毒性、细胞损伤程度、神经炎症,采用反转录定量聚合酶链反应(RT-qPCR)法分析相关基因表达,采用免疫印迹(Western blot)法分析相关蛋白表达。结果Sev能降低HT22细胞的活力,并增加HT22细胞中LDH,MDA,IL-6,TNF-α的含量,且呈剂量依赖性(P<0.05)。Dex能消除Sev对HT22细胞活力、神经毒性、氧化损伤程度和炎性反应的影响。Sev处理使HT22细胞中miR-204-5p的表达水平显著上升(P<0.05),而BDNF信使核糖核酸(mRNA)和蛋白表达水平均显著下降(P<0.05),且p-TrkB/TrkB水平显著下降(P<0.05);与Sev组相比,Dex+Sev组的miR-204-5p表达水平显著下降(P<0.05),而BDNF mRNA和蛋白表达水平均显著上升(P<0.05),且p-TrkB/TrkB水平显著上升(P<0.05)。miR-204-5p inhibitor能消除Sev诱导的神经毒性,而miR-204-5p mimic能逆转Dex对Sev诱导的神经毒性的保护作用。经TargetScan数据库分析显示,BDNF mRNA的3'非翻译区域中有与miR-204-5p结合的互补配对应答区域;双荧光素酶报告基因实验结果证实了miR-204-5p与BDNF的结合关系,且miR-204-5p能负调控HT22细胞中的BDNF mRNA和蛋白表达。BDNF siRNA能逆转Dex对Sev诱导的神经毒性的保护作用。结论Sev通过降低HT22细胞活力、增加LDH释放、促进氧化损伤和炎性反应来诱导神经毒性,而Dex能逆转这些作用,且Dex对Sev诱Objective To investigate the effect of dexmedetomidine(Dex)on sevoflurane(Sev)-induced oxidative damage and neuroinflammation in mouse HT22 hippocampal neuronal cells,and the possible regulatory mechanisms based on miR-204-5p/brain-derived neurotrophic factor(BDNF)/tyrosine receptor kinase B(TrkB)axis.Methods HT22 cells were pre-treated with 10µmol/L Dex for 2 h,and then treated with 3.4%Sev processing for 6 h.The miR-204-5p mimic,miR-204-5p inhibitor,BDNF siRNA or corresponding negative control were transfected into HT22 cells,and then were treated with Dex and/or Sev.CCK-8 method was used to detect cell viability,and lactate dehydrogenase(LDH)cytotoxicity assay kit,malondialdehyde(MDA)assay kit,interleukin-6(IL-6)enzyme-linked immunosorbent assay(ELISA)assay kit,and tumor necrosis factor-α(TNF-α)ELISA assay kit were used to detect neurotoxicity,cellular damage,and neuroinflammation,respectively.Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to analyze related gene expression,and Western blot was used to analyze related protein expression.Results Sev could reduce the viability of HT22 cells and increase the levels of LDH,MDA,IL-6,and TNF-αin HT22 cells in a dose-dependent manner(P<0.05).Dex could eliminate the effects of Sev on HT22 cell viability,neurotoxicity,degree of oxidative damage,and inflammatory response.Sev significantly up-regulated the expression level of miR-204-5p(P<0.05),significantly down-regulated the expression levels of BDNF messenger RNA(mRNA)and protein(P<0.05),and significantly reduced the level of p-TrkB/TrkB in HT22 cells(P<0.05).Compared with those in the Sev group,the expression level of miR-204-5p significantly down-regulated(P<0.05),the expression levels of BDNF mRNA and protein significantly up-regulated(P<0.05),and the p-TrkB/TrkB levels significantly increased in the Dex+Sev group(P<0.05).miR-204-5p inhibitor could eliminate Sev-induced neurotoxicity,while miR-204-5p mimic could reverse the protective effect of Dex on Sev-induced neurotoxicity.Tar

关 键 词：右美托咪定 七氟醚 氧化应激 海马神经元 神经炎症 微小核糖核酸 

分 类 号：R965[医药卫生—药理学] R971[医药卫生—药学]