中国药业2025，Vol.34Issue(7)：50-58,9.DOI:10.3969/j.issn.1006-4931.2025.07.011
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
摘要
Abstract
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 10 µmol/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-α in 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.TargetScan database analysis showed that there was a complementary paired response region in the 3'UTR of BDNF mRNA that bound to miR-204-5p.The results of the dual luciferase reporter gene assay confirmed the binding relationship between miR-204-5p and BDNF,and miR-204-5p could negatively regulate BDNF mRNA and protein expression in HT22 cells.BDNF siRNA could reverse the protective effect of Dex on Sev-induced neurotoxicity.Conclusion Sev induces neurotoxicity by reducing HT22 cell viability,increasing LDH release,promoting oxidative damage and inflammatory response,while Dex can reverse these effects,and the protective effect of Dex on Sev-induced neurotoxicity is realized through the miR-204-5p/BDNF/TrkB axis.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Key words
dexmedetomidine/sevoflurane/oxidative stress/hippocampal neurons/neuroinflammation/microRNAs分类
医药卫生引用本文复制引用
张云,康林莉..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[J].中国药业,2025,34(7):50-58,9.基金项目
2020年四川省医学(青年创新)科研课题项目[S20271]. （青年创新）
