摘要
目的:研究发现给予脑缺血再灌注损伤大鼠超短波(ultrashort wave,USW)治疗,可抑制高尔基体应激的重要参与分子——分泌途径衍生钙离子转运ATP酶1(secretory pathway Ca^(2+)-ATPase 1,SPCA1)表达的下降,减少高尔基体等细胞器的破坏和神经元细胞的凋亡,从而减轻脑缺血再灌注损伤。本研究旨在从细胞水平探讨USW对小鼠N2a细胞氧糖剥夺复氧(oxygen-glucose deprivation/reperfusion,OGD/R)损伤及SPCA1表达的影响。方法:将N2a细胞随机分为对照(Con)组、模型(OGD/R)组、USW组。Con组细胞正常培养,OGD/R组给予OGD/R处理,USW组给予OGD/R后用USW进行处理。在倒置光学相差显微镜下观察细胞形态;采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测细胞活性,双染法流式细胞术检测细胞凋亡,蛋白质印迹法检测SPCA1的表达。结果:Con组细胞多数呈梭形,轮廓清晰,贴壁良好;OGD/R组细胞皱缩,轮廓模糊,贴壁性变差,出现较多悬浮死细胞;与OGD/R组比较,USW组细胞形态和贴壁性改善,轮廓更清晰,死细胞减少。与Con组比较,OGD/R组细胞活性降低,凋亡率增高,SPCA1表达水平下调,差异均具有统计学意义(均P<0.001);与OGD/R组比较,USW组细胞活性增加,凋亡率降低,SPCA1表达水平上调,差异均具有统计学意义(P<0.01或P<0.001)。结论:USW可减轻细胞OGD/R后的损伤,其保护作用可能与上调SPCA1的表达有关。
Objective:Application of ultrashort wave(USW)to rats with cerebral ischemia and reperfusion injury could inhibit the decrease of expression of secretory pathway Ca^(2+)-ATPase 1(SPCA1),an important participant in Golgi stress,reduce the damage of Golgi apparatus and the apoptosis of neuronal cells,thereby alleviating cerebral ischemiareperfusion injury.This study aims to investigate the effect of USW on oxygen-glucose deprivation/reperfusion(OGD/R)injury and the expression of SPCA1 at the cellular level.Methods:N2a cells were randomly divided into a control(Con)group,an OGD/R group,and an USW group.The cells in the Con group were cultured without exposure to OGD.The cells in the OGD/R group were treated with OGD/R.The cells in the USW group were treated with USW after OGD/R.Cell morphology was observed under the inverted phasecontrast optical microscope,cell activity was detected by cell counting kit-8(CCK-8),apoptosis was detected by flow cytometry,and SPCA1 expression was detected by Western blotting.Results:Most of the cells in the Con group showed spindle shape with a clear outline and good adhesion.In the OGD/R group,cells were wrinkled,with blurred outline,poor adhesion,and lots of suspended dead cells appeared;compared with the OGD/R group,the cell morphology and adherence were improved,with clearer outlines and fewer dead cells in the USW group.Compared with the Con group,the OGD/R group showed decreased cell activity,increased apoptotic rate,and down-regulating SPCA1 expression with significant differences(all P<0.001);compared with the OGD/R group,the USW group showed increased cell activity,decreased apoptotic rate,and up-regulating SPCA1 expression with significant differences(P<0.01 or P<0.001).Conclusion:USW alleviates the injury of cellular OGD/R,and its protective effect may be related to its up-regulation of SPCA1 expression.
作者
唐金玲
王如蜜
樊永梅
张长杰
孔瑛
TANG Jinling;WANG Rumi;FAN Yongmei;ZHANG Changjie;KONG Ying(Department of Rehabilitation Medicine,Second Xiangya Hospital,Central South University,Changsha 410011,China)
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2023年第5期641-647,共7页
Journal of Central South University ：Medical Science
基金
湖南省自然科学基金(2020JJ5803)。
