摘要
目的探讨MAGI2-AS3/miR-374b-5p/HOXB7轴在脓毒症相关脑损伤(SE)发病中的作用。方法通过盲肠结扎穿孔(CLP)手术在成年C57BL/6 J雄性小鼠中诱导脓毒症。随机选择CLP小鼠用针对MAGI2-AS3的短干扰RNA(siRNA)或miR-374b-5p模拟物及相关对照转染。在术后8~12 d通过Morris水迷宫试验检测小鼠的空间学习能力和记忆能力。然后检测CLP小鼠的海马神经元凋亡指标。体外建立LPS诱导海马神经元损伤模型,分别用si-MAGI2-AS3或miR-374b-5p模拟物转染神经元,以确定海马神经元的增殖抑制和凋亡。使用双荧光素酶报告基因测定评估MAGI2-AS3、miR-374b-5p、HOXB7之间的相互作用。结果CLP小鼠在CLP手术后第3、7和14天显示MAGI2-AS3水平逐渐增加和miR-374b-5p水平逐渐降低,并伴有认知功能受损。MAGI2-AS3沉默和miR-374b-5p升高提高了CLP小鼠的空间学习能力和记忆能力,抑制了神经元凋亡。MAGI2-AS3沉默和miR-374b-5p升高增加了LPS处理的海马神经元的活力并减少了凋亡。MAGI2-AS3特异性结合miR-374b-5p,而miR-374b-5p靶向HOXB7。结论MAGI2-AS3沉默可能通过上调miR-374b-5p和下调HOXB7来抑制CLP小鼠海马神经元的凋亡。
Objective To explore the role of MAGI2-AS3/miR-374b-5p/HOXB7 axis in the pathogenesis of sepsis-associated encephalopathy(SE).Methods Sepsis was induced in adult C57BL/6J male mice by cecal ligation and perforation(CLP)surgery.Randomly selected CLP mice were transfected with short interfering RNAs(siRNAs)against MAGI2-AS3,or with miR-374b-5p mimics and related controls.Cognitive behavior was tested8 ~ 12 days after the surgery using a Morris water maze.Hippocampal neuronal apoptosis indicators in CLP mice were then detected.The hippocampal neuron damage model was induced by LPS.Next,the neurons were,respectively,transfected with si-MAGI2-AS3 or miR-374b-5p mimics to determine the proliferation inhibition and apoptosis of hippocampal neurons.The interactions between MAGI2-AS3,miR-374b-5p and HOXB7 were assessed using dual luciferase reporter gene assay.Results CLP mice showed progressive increases in MAGI2-AS3 levels and progressive decreases in miR-374b-5p levels on the 3rd,7th,and 14th after CLP surgery,accompanied by impaired cognitive function.MAGI2-AS3 silencing and miR-374b-5p elevation improved spatial learning ability and memory ability,and depressed neuronal apoptosis in CLP mice.MAGI2-AS3 silencing and miR-374b-5p elevation increased viability and limited apoptosis of LPS-treated hippocampal neurons.MAGI2-AS3 specifically bound to miR-374b-5p,and miR-374b-5p targeted HOXB7.Conclusion MAGI2-AS3 silencing inhibits the apoptosis of hippocampal neurons in CLP mice by up-regulating miR-374b-5p and down-regulating HOXB7.
作者
黄巍
张庆祥
罗丹
冯彰凤
HUANG Wei;ZHANG Qingxiang;LUO Dan;FENG Zhangfeng(Department of Critical Care Medicine,Guilin Peo-ple′s Hospital,Guilin 541002,China)
出处
《实用医学杂志》
CAS
北大核心
2022年第7期815-820,共6页
The Journal of Practical Medicine
基金
桂林市科学与技术开发计划项目(编号:20190218-7-10)
2020年广西壮族自治区医药卫生科研课题(编号:20200037)。