Objectives: Application of ultrashort wave (USW) to rats with cerebral ischemia and reperfusion injury could inhibit the decrease of expression of secretory pathway Ca2+-ATPase 1 (SPCA1), an important participant in Golgi stress, reduce the damage of Golgi apparatus and the apoptosis of neuronal cells, thereby alleviating cerebral ischemia-reperfusion 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 phase-contrast 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).
Conclusions: USW alleviates the injury of cellular OGD/R, and its protective effect may be related to its up-regulation of SPCA1 expression.
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 Ca2+-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。.
Keywords: Golgi apparatus stress; apoptosis; oxygen-glucose deprivation/reperfusion; secretory pathway Ca 2+-ATPase 1; ultrashort wave.