高血糖对大鼠局灶性脑缺血再灌注时小胶质细胞活化的影响

发布时间：2019-02-13 05:21

【摘要】：目的观察高血糖对大鼠局灶性脑缺血再灌注时小胶质细胞活化的影响。方法健康雄性SD大鼠80只,随机分为正常血糖假手术组(NG sham组,n=5)、高血糖假手术组(HG sham组,n=5)、正常血糖脑缺血再灌注组(NG手术组,n=35)和高血糖脑缺血再灌注组(HG手术组,n=35)。采用链脲佐菌素(STZ)腹腔注射法制作SD大鼠1型糖尿病模型,大脑中动脉阻塞法(MCAO)建立大鼠脑缺血再灌注模型,应用小胶质细胞特异性标志蛋白Iba-1免疫组化标记小胶质细胞,观察NG组和HG组大脑中动脉阻塞30min,再灌注0.5、3、6h以及1、3、7、14d大鼠(每时间点5只)室周带和尾状核区小胶质细胞的变化;采用Iba-1和核增殖抗原(PCNA)免疫荧光双标法检测小胶质细胞的增殖变化。结果脑缺血再灌注后,可见大鼠脑组织明显水肿,呈网格状,HE染色变淡,神经元肿胀,胞质空泡状,胞核固缩,并可见炎细胞浸润,HE染色结果显示HG手术组与NG手术组比较脑损伤更为明显。脑缺血再灌注后第3天,免疫组化染色可见梗死周边区、梨状皮质和躯体感觉皮质的小胶质细胞明显活化,于第7天达到峰值,且其活化状态可以持续到再灌注第14天。免疫荧光双标染色结果显示脑缺血再灌注后小胶质细胞数量增加与其增殖有关,其增殖程度同样是在缺血再灌注后第3天增加,第7天时达高峰。与NG手术组比较,HG手术组小胶质细胞的活化及增殖较弱(P0.05),但均明显高于各自的假手术组(P0.05)。结论高血糖导致的缺血后脑组织小胶质细胞活化、增殖抑制可能参与了高血糖加重缺血性脑损伤的过程。
[Abstract]:Objective to observe the effect of hyperglycemia on the activation of microglia during focal cerebral ischemia reperfusion in rats. Methods 80 healthy male SD rats were randomly divided into normal blood glucose sham-operation group (NG sham group, n = 5), hyperglycemia sham operation group (HG sham group, n = 5), normal blood glucose cerebral ischemia reperfusion group (NG operation group), hyperglycemia sham operation group (HG sham group, n = 5), and normal blood glucose cerebral ischemia reperfusion group (NG operation group). NM35) and hyperglycemic cerebral ischemia-reperfusion group (HG operation group, nong35). The model of type 1 diabetes in SD rats was established by intraperitoneal injection of streptozotocin (STZ) and middle cerebral artery occlusion (MCAO). Microglial cells were labeled with microglia specific marker protein Iba-1. The middle cerebral artery occlusion was observed for 30 min in NG group and HG group. Changes of microglia in periventricular zone and caudate nucleus of rats (5 rats per time point) at 7d 14; Iba-1 and (PCNA) were used to detect the proliferation of microglia. Results after cerebral ischemia-reperfusion, the brain tissue of rats showed obvious edema, reticular shape, light staining of HE, swelling of neurons, vacuolation of cytoplasm, pyknosis of nucleus and infiltration of inflammatory cells. HE staining showed that brain injury was more obvious in HG group than in NG group. On the 3rd day after cerebral ischemia-reperfusion, the microglia in the infarct area, piriform cortex and somatosensory cortex were obviously activated, and reached the peak on the 7th day, and the activation state could continue until the 14th day of reperfusion. The results of double immunofluorescence staining showed that the increase of the number of microglia after cerebral ischemia-reperfusion was related to the proliferation of microglia. The proliferative degree of microglia increased on the 3rd day after ischemia-reperfusion and reached its peak on the 7th day. The activation and proliferation of microglia in HG group were weaker than those in NG group (P0.05), but they were significantly higher than those in sham-operated group (P0.05). Conclusion the activation of microglia and the inhibition of proliferation may be involved in the process of hyperglycemia exacerbating ischemic brain injury.
【作者单位】：宁夏医科大学基础医学院病理学系;宁夏医科大学法医司法鉴定中心;
【基金】：宁夏医科大学校级课题(XY2017002);宁夏医科大学基础医学西部一流学科建设项目~~
【分类号】：R587.2;R741

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