用于脑缺血小鼠海马SOD1表达的改进灌注固定法

常规灌注固定法多用于兔和大鼠等较大动物,并存在一些不足。改进了灌注固定法流程、灌注溶液的配方、流速、用量以及灌注装置,将其用于在显微操作下制备的缺血再灌注C57BL/6N小鼠模型,并对其海马进行H.E染色和免疫组织化学SOD1基因表达。结果显示,改进的灌注固定法使组织切片结构更加清晰,海马免疫阳性神经元定位于胞浆。缺血再灌注组(24hI/R)海马神经元SOD1表达比假手术对照组(sham-o)减少,而高压氧治疗组(24hHBO)SOD1表达有所恢复。表明改进的灌注固定法用于缺血再灌注C57BL/6N小鼠海马SOD1基因表达效果良好,结果可靠。实验结果提示,高压氧的治疗机制之一可能是通过增加SOD1基因表达而实现的。

Application of Modified Perfusion Fixation in the Study on the Expression of Copper/zinc Superoxide Dismutase in Mouse Hippocampus after Transient Forebrain Ischemia

Routine perfusion fixation has been used in rabbits or rats,but this method has some shortage.An improved perfusion fixation method was used in C57 BL/6N mouse ischemia-perfusion model.Samples were observed with H.E staining and immunohistochemistry staining to study the expression of copper/zinc superoxide dismutase(SOD1) in hippocampus of C57 BL/6N mouse after transient forebrain ischemia.Results showed that the tissue structure become clearer when the modified perfusion fixation method was used:masculine neurons of hippocampus were located in the cytoplasm.The SOD1 expression level of the ischemia-reperfusion(24 h I/R) group was lower than that of sham-operated(sham-o) control,but that of hyperbaric oxygen treatment(24 h HBO) group almost recovered.These results illustrated that the modified perfusion fixation was good for immunohistochemistry in C57 BL/6N mouse hippocampus after transient forebrain ischemia-reperfusion.The results also indicated that one of the mechanisms of HBO therapy may be achieved by the increase of SOD1expression level.