收稿日期: 2023-04-09
网络出版日期: 2024-03-08
基金资助
广西自然科学基金面上项目(2020GXNSFAA297161);第七批全国老中医专家学术经验继承工作(国中医药人教函〔2022〕76号);广西中医临床优秀人才项目(桂中医药科教发［2022］14号);刘泰广西名中医传承工作室建设项目(桂中医药科教发［2021］6号)
The mechanism of target regulating of miR⁃421 to Menin/Caspase⁃3 pathway for depression
Received date: 2023-04-09
Online published: 2024-03-08
目的 探讨miR-421影响抑郁症发生发展的作用机制。 方法 采取单次腹腔注射脂多糖（LPS）方法建立抑郁大鼠模型，采用糖水偏好度测试和旷场实验进行抑郁行为检测。通过miRNA微阵列芯片和RT-PCR分析miR-421在抑郁大鼠海马组织中的表达量，运用TargetScan数据库和mi RDB数据库进行预测miR-421的靶基因，采用双荧光素酶报告基因实验观察其与靶基因的结合情况，观察过表达和抑制miR-421对靶基因的影响，随后过表达和抑制靶基因，观察其对下游因子的影响，最终探究miR-421影响抑郁症的相关机制。 结果 miRNA微阵列芯片和RT‐PCR检测表明miR-421在抑郁大鼠海马组织中呈高表达（P < 0.001），抑制miR-421的表达可显著恢复抑郁大鼠的体重和运动能力（P < 0.001）。TargetScan数据库预测得到Menin与miR-421存在结合靶点，双荧光素酶报告基因实验表明Menin与miR-421具有相互作用；当miR-421过表达时，Menin表达量会下调（P < 0.001），相反，当抑制miR-421表达时，Menin表达量会上调（P < 0.001）。qPCR检测提示，Menin下游因子Caspase-3、NF-κB在抑郁大鼠模型海马组织中的表达显著提高（P < 0.001），IL-1β在抑郁大鼠模型海马组织中的表达明显提高（P < 0.01），当抑制Menin表达时，Caspase-3、NF-κB、IL-1β表达量会升高（P < 0.001），当过表达Menin时，Caspase-3、NF-κB、IL-1β表达量则降低（P < 0.001）。 结论 抑制miR-421表达可升高Menin表达，降低Caspase-3含量，减少神经炎症反应，从而改善抑郁症状。
刘永辉 , 谭庆晶 , 陈清 , 韦理萍 , 杨俊威 , 杨侃 , 高玉广 . miR-421靶向调控Menin/Caspase-3影响抑郁症的机制[J]. 实用医学杂志, 2024 , 40(4) : 453 -459 . DOI: 10.3969/j.issn.1006-5725.2024.04.003
Objective To explore the mechanism of miR-421 affecting the occurrence and development of depression. Methods A depressive rat model was established by single intraperitoneal injection of lipopolysaccharide （LPS）， and depressive behavior was detected by glucose preference test and open-field test. miRNA microarray chips and RT-PCR were used to analyze the expression level of miR-421 in hippocampus of the depressed rats. TargetScan database and mi RDB database were used to predict the target genes of miR-421. Dual luciferase reporter gene assay was used to observe the binding of miR-421 to the target genes. The impact of over-expression and inhibition of miR-421 on target genes was observed， then the influence of over-expression and inhibition of target genes on downstream factors was observed， and the related mechanism of miR-421 on depression was explored. Results miRNA microarray chips and RT-PCR assay showed that miR-421 was highly expressed in the hippocampus of the depressed rats （P < 0.001）， Inhibition of miR-421 expression could significantly restore the body weight and exercise ability of the depressed rats （P < 0.001）. Binding targets of Menin and miR-421 were predicted by TargetScan database， and interaction between Menin and miR-421 was demonstrated by dual-luciferase reporter gene assay. Menin expression was down-regulated while miR-421 was overexpressed （P < 0.001）， whereas it was up-regulated as miR-421 was inhibited （P < 0.001）. qPCR indicated that expressions of Caspase-3 and NF-κB in the hippocampus of the depressed rats was significantly increased （P < 0.001）， and IL-1β expression in the hippocampus was significantly increased （P < 0.01）. When the expression of Menin was inhibited， the expressions of Caspase-3， NF-κB and IL-1β were increased （P < 0.001）， while the expressions of Caspase-3， NF-κB and IL-1β were decreased when Menin was overexpressed （P < 0.001）. Conclusions Inhibition of miR-421 expression can increase Menin expression， decrease Caspase-3 content， and reduce neuroinflammatory response， thereby improving depressive symptoms.
Key words： depression; miR-421; menin; caspase 3; animal experiments; mechanism
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