【摘要】：目的高血糖能夠加重腦缺血再灌注損傷，但具體的途徑和分子機(jī)制尚不清楚，本研究以高血糖局灶性腦缺血再灌注損傷大鼠作為模型，探討高血糖加重腦缺血再灌注損傷中神經(jīng)元及星形膠質(zhì)細(xì)胞氧化損傷的作用。 方法將雄性SD大鼠隨機(jī)化分為正常血糖腦缺血再灌注組（簡稱正常血糖組）、糖尿病高血糖腦缺血再灌注組（簡稱糖尿病組）以及假手術(shù)對照組（簡稱假手術(shù)組），采用鏈脲佐菌素（Streptozotocin,STZ,55mg/kg）制備Ⅰ型糖尿病大鼠模型，糖尿病高血糖形成后3d內(nèi)實(shí)施MCAO缺血30min，再灌注1d，3d，7d，14d。分別進(jìn)行組織學(xué)、8羥基脫氧鳥苷（8-hydroxy-2’deoxyguanosine，8-OHdG）免疫組織化學(xué)、NeuN和8-OHdG免疫熒光雙標(biāo)記及GFAP和8-OHdG免疫熒光雙標(biāo)記技術(shù)，對比觀察各組神經(jīng)元及星形膠質(zhì)細(xì)胞的損傷及其與8-OHdG的關(guān)系。 結(jié)果：組織學(xué)顯示，正常血糖組再灌注1d腦組織出現(xiàn)水腫，糖尿病組較正常血糖組有所加重，出現(xiàn)較多的固縮神經(jīng)元，再灌注3d腦組織水腫進(jìn)一步加重，固縮神經(jīng)元進(jìn)一步增多,可見多數(shù)細(xì)胞溶解壞死，存留泡沫樣結(jié)構(gòu)，同樣，糖尿病組表現(xiàn)更為嚴(yán)重；再灌注7d正常血糖組神經(jīng)元固縮和腦水腫明顯減輕，糖尿病組仍可見少數(shù)神經(jīng)元固縮和腦水腫；再灌注14d正常血糖組神經(jīng)元固縮和腦水腫消失，膠質(zhì)細(xì)胞增加，糖尿病組可見輕度腦水腫。免疫組化顯示，再灌注3d，正常血糖組及糖尿病組8-OHdG陽性細(xì)胞數(shù)量均明顯增加，高血糖組8-OHdG陽性細(xì)胞數(shù)量明顯高于正常血糖組（P0.05），再灌注7d和14d8-OHdG免疫陽性細(xì)胞明顯減少，但仍多于假手術(shù)組（P0.05）；免疫熒光雙標(biāo)記提示，，再灌注3d時(shí)，正常血糖組及糖尿病組8-OHdG陽性的神經(jīng)元均達(dá)高峰，且糖尿病組的8-OHdG陽性神經(jīng)元明顯高于正常血糖組（P0.05），再灌注7d和14d時(shí)，8-OHdG陽性的神經(jīng)元有所減少；且正常血糖組再灌注3d8-OHdG陽性的星形膠質(zhì)細(xì)胞開始增加，再灌注7d時(shí)達(dá)高峰，糖尿病組較正常血糖組雙標(biāo)陽性細(xì)胞明顯增加（P0.05），再灌注14d雙標(biāo)陽性細(xì)胞數(shù)有所下降,但仍多于假手術(shù)組。 結(jié)論：（1）糖尿病高血糖可以加重腦缺血再灌注損傷，使局灶性腦缺血梗死區(qū)及其周圍腦組織水腫加重，梗死區(qū)擴(kuò)大，神經(jīng)元變性和凋亡增加，恢復(fù)時(shí)間延長；（2）腦缺血再灌注引起中樞神經(jīng)系統(tǒng)氧化損傷，局灶性腦缺血30min，再灌注1天氧化損傷明顯增加，再灌注3天時(shí)達(dá)到高峰，再灌注7天氧化損傷明顯減少；糖尿病高血糖能夠明顯加重再灌注1天和3天時(shí)的氧化損傷，再灌注7天氧化損傷減輕的幅度明顯大于正常血糖組；（3）局灶性腦缺血30min，再灌注1天和3天氧化損傷明顯增加，氧化損傷主要發(fā)生在神經(jīng)元；糖尿病高血糖能夠明顯加重神經(jīng)元的氧化損傷，星形膠質(zhì)細(xì)胞氧化損傷較輕微。
[Abstract]:Objective hyperglycemia can aggravate cerebral ischemia-reperfusion injury, but the specific pathway and molecular mechanism are not clear. In this study, focal cerebral ischemia-reperfusion injury induced by hyperglycemia in rats was used as model. To investigate the effect of hyperglycemia on oxidative injury of neurons and astrocytes in cerebral ischemia reperfusion injury. Methods male Sprague-Dawley rats were randomly divided into normal blood glucose cerebral ischemia-reperfusion group, diabetic hyperglycemic cerebral ischemia reperfusion group and sham operation control group. The rat model of type 1 diabetes mellitus was established by using streptozotocin (STZ) 55 mg / kg. Diabetic hyperglycemia was performed within 3 days after MCAO ischemia for 30 minutes and reperfusion for 1 day for 3 days and 7 days for 14 days. Immunohistochemical staining of 8-hydroxy-2-deoxyguanosine 8-OHdG (Neun and 8-OHdG) and double labeling of GFAP and 8-OHdG were performed respectively. The damage of neurons and astrocytes and their relationship with 8-OHdG were observed. Results: histologically, there were edema in brain tissue of normal blood glucose group on the 1st day after reperfusion, more pyknotic neurons appeared in diabetes group than in normal blood glucose group, and the edema of brain tissue was further aggravated at 3 days after reperfusion. The pyknotic neurons increased further, most of the cells dissolved and necrotized, and the foam structure remained. Similarly, the diabetic group showed more serious symptoms, while the normal blood glucose group on the 7th day after reperfusion significantly alleviated the pyknosis and brain edema. There were still a few neurons pyknosis and brain edema in the diabetic group, while the neuron pyknosis and brain edema disappeared in the normal blood glucose group on the 14th day after reperfusion, the glial cells increased, and mild brain edema was observed in the diabetic group. Immunohistochemistry showed that the number of 8-OHdG positive cells in normal blood glucose group and diabetic group was significantly increased on the 3rd day after reperfusion, and the number of 8-OHdG positive cells in hyperglycemia group was significantly higher than that in normal blood glucose group (P0.05), and the number of 14d8-OHdG immunoreactive cells and 14d8-OHdG immunoreactive cells in hyperglycemia group decreased significantly at 7 days after reperfusion. But it was still more than sham operation group (P0.05), immunofluorescence double labeling showed that the 8-OHdG positive neurons in normal blood glucose group and diabetes group reached the peak at 3 days after reperfusion. The 8-OHdG positive neurons in the diabetic group were significantly higher than those in the normal blood glucose group (P0.05), and the number of 8-OHdG positive neurons decreased on the 7th and 14th days after reperfusion, and the number of 3d8-OHdG positive astrocytes in the normal blood glucose group began to increase and reached the peak at the 7th day after reperfusion. The number of double labeled positive cells in diabetes group was significantly higher than that in normal blood glucose group (P0.05), and the number of double labeled positive cells decreased on the 14th day after reperfusion, but still more than that in sham operation group. Conclusion: (1) Diabetic hyperglycemia can aggravate cerebral ischemia-reperfusion injury, aggravate edema, enlarge infarct area, increase neuronal degeneration and apoptosis, and prolong recovery time; (2) oxidative injury of central nervous system was induced by cerebral ischemia-reperfusion. After 30 min of focal cerebral ischemia, the oxidative injury increased significantly on 1 day after reperfusion, reached the peak at 3 days after reperfusion, and decreased significantly on 7 days after reperfusion. Diabetic hyperglycemia could significantly aggravate oxidative injury on 1 and 3 days after reperfusion, and the extent of oxidative damage at 7 days after reperfusion was significantly higher than that in normal blood glucose group, (3) oxidative damage was significantly increased at 30 min after focal cerebral ischemia, 1 day and 3 days after reperfusion. The oxidative damage occurred mainly in neurons, and diabetic hyperglycemia significantly aggravated the oxidative damage of neurons, while the oxidative damage of astrocytes was slight.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743.3

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