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

作　　者：王漫漫 兰瑞[2] 张勇[3] 付雪琴[1] 邹旭欢 王玮玮 李泓宇 唐琛 刘双 WANG Manman;LAN Rui;ZHANG Yong;FU Xueqin;ZOU Xuhuan;WANG Weiwei;LI Hongyu;TANG Chen;LIU Shuang(Department of Traditional Chinese Medicine,Fisrt Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450046,China;Encephalopathy Center,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099,China;The Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]河南中医药大学第一临床医学院,河南郑州450046 [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

出　　处：《中国病理生理杂志》2024年第1期126-133,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81973618,No.81503422);河南省自然科学基金资助项目(No.202300410399)。

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Neu N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。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 deprivation/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 concentration.The HT22 cells were divided into two groups:control and model(OGD/R).Cellular morphological changes were observed using an inverted microscope.The levels of IL-1β,IL-6 and TNF-αin the supernatant were quantified by ELISA.Ultrastructural changes were examined by transmission electron microscopy.Immunofluorescence staining was used to detect neuron markers Neu N 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-αlevels(P<0.05).Immunofluorescence intensity and protein levels of Neu N,NF200 and MAP2 were notably reduced(P<0.05).Treatment with XXMD improved cell morphology,ultrastructure and survival rate(P<0.05),and decreased inflammatory 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 Neu N,NF200 and MAP2(P<0.05).CONCLUSION:Xiaoxuming decoction may mitigate OGD/R-induced injury,potentially by inhibiting inflammatory responses and enhancing 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 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]