| Objective:Further verify the mice cerebral ischemia and reperfusion injury in experimental animal models of relevance to observe the cerebral ischemia-reperfusion injury in mice at different time points after the intervention given to the effect of mannitol, verification on the cerebral ischemia-reperfusion injury mannitol intervention Best time of the study.Methods:36 healthy Kunming mice, male and female, regardless of body weight 30±5g, were randomly divided into 6 groups, each group 6. A:sham group, B group:ischemia-reperfusion injury group (control group), C group:mannitol immediately after reperfusion in the intervention group, D group:30min after reperfusion mannitol treatment group, E group:Reperfusion 1h after mannitol treatment group, F group:1.5h after reperfusion mannitol treatment group. Dose of mannitol in the intervention group according to 1g/kg given intravenous injection of 20% mannitol injection. Reported in mice with early bilateral carotid artery ligation model of cerebral ischemia, reperfusion in each group after 24h fast craniotomy the brain. Specimens placed in 4% paraformaldehyde solution fixed 24h after the brain tissue obtained after the optic chiasm, conventional paraffin-embedded, coronal slices, slice thickness of 5μm, according to the brain tissue samples need to be addressed, some of the hematoxylin-eosin staining (HE staining), to verify the completion of senior Guo Wenyi previous experimental results, reperfusion injury were observed at different times after intervention on the hippocampal CA1 area of mannitol neuronal survival in cell number, degeneration cell rate, histological and behavioral changes in mice Effects. Another part of the terminal deoxynucleotidyl transferase-mediated dUTP nick end tag of the original index bits (TUNEL) detected, observed on nerve cell apoptosis.Result:HE dyed observation:Sham group seahorses CA1 area layer 4 organizational structure integrity, distinct.5 layers pyramidal cells clearly visible and closely-arranged, cytoplasmic homogeneous red dye, nuclear is big and round and nuclear membrane integrity, nucleoli clear. Cell layer to the heart side the stroma is typical fiber bundle sample structure, orderly rows. Ischemia reperfusion group layer 4 organizational structure variable density, interstitial edema. Even just visible 2-3 layers pyramidal cells, normal cone cell count, obviously, obviously, a lot of reducing degeneration necrosis cells, cells swelling, the cytoplasm, free bubble formed cytoplasm eosinophilic enhancement, the necrotic the nuclei are fusiform or triangle become pyknotic, deep dyed blue purple. Cell edema obvious, a crumb. Cell layer to the heart side interstitial fiber bundle sample structure arrangement messy, interrupt irregular. C mice surviving cells in hippocampus CA1 area was significantly higher than B group (p<0.05), degeneration of cells was significantly lower than the control group, ischemia-reperfusion (p <0.05), but not with the A group Significant difference (p>0.05), D, E, F group and B group showed no significant difference (p>0.05).TUNEL staining: HE stain morphology consistent with the sham group sections occasionally positive cells (apoptosis in normal cells also.) Ischemia-reperfusion injury seen in much larger group of slices of different sizes, different patterns of brown positive cells (apoptotic cells increased). C hippocampal CA1 area of mice apoptosis index (AI) was significantly lower than B group (p<0.05), with the A group was not significantly different (p>0.05), D, E, F hippocampal CA1 area of mice were Shows more positive cells over time gradually increased apoptosis, F group and B group was no significant difference (p>0.05).Conclusion:From the morphological point of view, mannitol early intervention can significantly reduce ischemic reperfusion injury, resumed 30 minutes after infusion of mannitol given intervention, ischemic nerve cell swelling has been formed, a variety of cell damage mechanisms are activated Showed an increase in apoptotic cells, and then given mannitol to the basic treatment is invalid. The experimental results suggest that the rescue of clinical cardiac arrest, heart rate recovery in the autonomous should start at the same time that mannitol dehydration treatment.
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