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Astrocyte Changes Involved in Cerebral Ischemia and Reperfusion Injury under Hyperglycemia in Rats

Author: MaXiuPing
Tutor: ZhangJianZhong；JingLi
School: Ningxia Medical University
Course: Pathology and pathophysiology
Keywords: Brain Ischemia / reperfusion Astrocytes Apoptosis Extracellular signal-regulated kinase 1/2 Cysteine ??aspartate - specific proteolytic enzyme -3
CLC: R741
Type: Master's thesis
Year: 2011
Downloads: 37
Quote: 0
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Abstract


The purpose of high blood sugar can aggravate cerebral ischemia reperfusion neuronal injury, astrocytes unclear. In this study, a high blood sugar whole brain ischemia reperfusion in rats as a model to explore the high blood sugar cerebral ischemia reperfusion injury in astrocytes role and molecular mechanisms. Methods Male SD rats were randomly divided into sham control group (sham group): diabetic hyperglycemia, normal blood sugar cerebral ischemia reperfusion group (referred to as the normal glucose group), the diabetic hyperglycemia cerebral ischemia reperfusion group (the high glycemic group). The streptozotocin (Streptozotocin, STZ) Preparation of type I diabetes rat model, by bilateral carotid artery occlusion and femoral artery phlebotomy, the establishment of global cerebral ischemia 15min, reperfusion 3h, 6h model. Histopathological observation, in situ end deoxynucleotidyl transferase mediated deoxyuridine triphosphate (dUTP) labeling method (TUNEL), immunofluorescence double labeling, Western blot method (Western blot) technology, respectively, contrast observed astrocyte apoptosis and phosphorylation of MEK1 / 2, phosphorylated ERK1 / 2 and Cleaved Caspase-3 protein expression. The results of histological examination showed the hyperglycemia group cerebral ischemia reperfusion 6h, swelling of neurons, significant cerebral edema. TUNEL double labeling in the sham group found no positive nerve cells; hyperglycemia group cerebral ischemia reperfusion 3h hippocampal CA1 region of TUNEL-positive nerve cells is significantly higher than normal blood glucose group (P lt; 0.05). Were double-labeled by GFAP and TUNEL fluorescence immunoassay visible in normal glucose group cerebral ischemia reperfusion 3h and 6h CA1 and cingulate cortex occasionally TUNEL-positive astrocytes, hyperglycemia group brain missing blood reperfusion 6h, hippocampal CA1 region of TUNEL-positive astrocytes significantly more than normal blood glucose group (P lt; 0.05). Cleaved Caspase-3 immune markers visible in the hyperglycemia group and normal glucose group, hippocampal CA1 and cingulate cortex visible positive astrocytes, the hyperglycemia group cerebral ischemia and reperfusion 3h and 6h, Cleaved Caspase-3 positive The astrocytes quantity than normal blood glucose group (P lt; 0.05). Western Blot analysis, the hyperglycemia group cerebral ischemia reperfusion 3h and 6h of cleaved caspase-3 relative gray values ??increased in the hippocampus compared with normal glucose group was statistically significant (P lt; 0.05). Astrocyte GFAP protein expression can be seen, compared with normal blood sugar, high blood sugar group can be seen in the hippocampal CA1 region and the cingulate cortex astrocyte cell bodies larger processes and thick. P-MEK1 / 2 immune double labeling visible, visible in normal glucose tolerance and hyperglycemia group P-MEK1 / 2 positive astrocytes; hyperglycemia group ischemia-reperfusion 3h, 6h positive glial cells in the hippocampal CA1 region than normal blood glucose group (P lt; 0.05); the hyperglycemia groups cingulate cortex positive astrocytes in ischemia-reperfusion 3h significant difference (P lt; 0.05) compared with normal glucose tolerance. Western Blot display, hyperglycemia group of brain ischemia and reperfusion 3h and 6h hippocampus P-MEK1 / 2 relative gray values ??(respectively 2.087 ± 0.147,1.780 ± 0.091) was significantly higher than normal blood glucose group (P lt; 0.05) . Phosphorylated ERK1 / 2 was not found in the sham group positive cells in the hippocampal CA1 and cingulate cortex hyperglycemia group cerebral ischemia and reperfusion a 3h-positive astrocytes significantly more than normal blood glucose group (P lt; 0.05). Western Blot analysis showed that the hyperglycemia group cerebral ischemia reperfusion 3h and 6h hippocampus P-ERK1 / 2 relative gray value is higher than normal blood glucose group (P lt; 0.05). Conclusion diabetes high blood sugar condition, cerebral ischemia and reperfusion three hours to be able to increase apoptosis in experimental rat hippocampal CA1 neurons. The hyperglycemia brain ischemia and reperfusion can cause astrocyte activation and increased apoptosis in experimental rat hippocampal CA1 and cingulate cortex. Apoptosis astrocytes increased late in neurons, may be related to the biological function of astrocytes. Hyperglycemia cerebral ischemia and reperfusion can be further raised the activation of ERK1 / 2 signaling pathway, hyperglycemia cerebral ischemia-reperfusion injury hyperphosphorylation of ERK1 / 2 signaling pathway may be mediated astrocyte withered death and activation.

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