中国病理生理杂志2024,Vol.40Issue(1) :126-133.DOI:10.3969/j.issn.1000-4718.2024.01.016

小续命汤对OGD/R诱导HT22细胞损伤后突触可塑性的影响

Effect of Xiaoxuming decoction on OGD/R-induced synaptic plasticity in HT22 cells

王漫漫 兰瑞 张勇 付雪琴 邹旭欢 王玮玮 李泓宇 唐琛 刘双
中国病理生理杂志2024,Vol.40Issue(1) :126-133.DOI:10.3969/j.issn.1000-4718.2024.01.016

小续命汤对OGD/R诱导HT22细胞损伤后突触可塑性的影响

Effect of Xiaoxuming decoction on OGD/R-induced synaptic plasticity in HT22 cells

王漫漫 1兰瑞 2张勇 3付雪琴 4邹旭欢 4王玮玮 4李泓宇 4唐琛 4刘双4
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作者信息

  • 1. 河南中医药大学第一临床医学院, 河南 郑州 450046;河南中医药大学第一附属医院脑病中心,河南 郑州 450099
  • 2. 河南中医药大学第一附属医院脑病中心,河南 郑州 450099
  • 3. 郑州大学第三附属医院中医科, 河南 郑州 450052
  • 4. 河南中医药大学第一临床医学院, 河南 郑州 450046
  • 折叠

摘要

目的:探讨小续命汤(XXMD)对缺血性脑卒中后脑缺血再灌注损伤突触可塑性的影响.方法:体外建立小鼠海马神经元(HT22细胞)氧糖剥夺/复氧(OGD/R)模型,模拟脑缺血性脑卒中后海马神经元缺血再灌注损伤.采用CCK-8法检测HT22细胞活力,筛选XXMD最佳作用浓度.将HT22细胞随机分为对照组、OGD/R组和OGD/R+XXMD组,倒置荧光显微镜观察各组细胞形态变化,ELISA法测定细胞上清液中炎症因子IL-1β、IL-6和TNF-α水平,透射电镜观察细胞超微结构改变,免疫荧光染色检测神经元标志物NeuN及突触相关蛋白NF200和MAP2的荧光强度,Western blot检测各组细胞中NF200和MAP2蛋白表达水平.结果:XXMD在浓度为100 mg/L时细胞活力最强(P<0.05).与对照组相比,模型组细胞变圆皱缩,线粒体呈现肿胀甚至空泡样改变,活力明显下降(P<0.05),IL-1β、IL-6和TNF-α含量升高(P<0.05),NF200和MAP2的荧光强度及蛋白表达水平显著降低(P<0.05).而XXMD可改善细胞形态及超微结构,提升生存率(P<0.05),降低炎症因子水平(P<0.05).免疫荧光染色及Western blot结果显示,与模型组相比,OGD/R+XXMD组可以增强NF200和MAP2平均荧光强度(P<0.05),提升蛋白表达水平(P<0.05).结论:XXMD可以减轻OGD/R诱导的HT22细胞损伤,其机制可能涉及减轻炎症反应和增强突触可塑性.

Abstract

AIM:To explore the protective effect of Xiaoxuming decoction(XXMD)on synaptic plasticity in the context of cerebral ischemia-reperfusion injury following ischemic stroke.METHODS:An oxygen-glucose depriva-tion/reoxygenation(OGD/R)model was employed in vitro using mouse hippocampal neurons(HT22 cells)to simulate ischemia-reperfusion injury.Cell viability was assessed using a CCK-8 assay to determine the optimal XXMD concentra-tion.The HT22 cells were divided into two groups:control and model(OGD/R).Cellular morphological changes were ob-served using an inverted microscope.The levels of IL-1β,IL-6 and TNF-α in the supernatant were quantified by ELISA.Ultrastructural changes were examined by transmission electron microscopy.Immunofluorescence staining was used to de-tect neuron markers NeuN and synaptic proteins NF200 and MAP2.The protein levels of NF200 and MAP2 were analyzed by Western blot.RESULTS:The highest cell survival rate occurred at an XXMD concentration of 100 mg/L(P<0.05).Compared with control group,the cells in model group exhibited round shape and shrinkage,mitochondrial swelling or vacuolization,and a marked decrease in survival rate.There were significant increases in IL-1β,IL-6 and TNF-α levels(P<0.05).Immunofluorescence intensity and protein levels of NeuN,NF200 and MAP2 were notably reduced(P<0.05).Treatment with XXMD improved cell morphology,ultrastructure and survival rate(P<0.05),and decreased in-flammatory factor levels(P<0.05).Compared with model group,the cells in OGD/R+XXMD group showed significantly increased immunofluorescence intensity and protein levels of NeuN,NF200 and MAP2(P<0.05).CONCLUSION:Xiaoxuming decoction may mitigate OGD/R-induced injury,potentially by inhibiting inflammatory responses and enhanc-ing synaptic plasticity.

关键词

小续命汤/氧糖剥夺/HT22细胞/缺血性脑卒中/突触可塑性

Key words

Xiaoxuming decoction/traditional medicinal formula/OGD/R/HT22/ischemic stroke/synap-tic plasticity

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基金项目

国家自然科学基金资助项目(81973618)

国家自然科学基金资助项目(81503422)

河南省自然科学基金资助项目(202300410399)

出版年

2024
中国病理生理杂志
中国病理生理学会

中国病理生理杂志

CSTPCD北大核心
影响因子:1.065
ISSN:1000-4718
参考文献量2
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