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Effects of Lithium Chloride on Neuronal Apoptosis and Expression of Glycogen Synthase Kinase (GSK)-3beta of Hippocampal CA1 Region after Transient Focal Cerebral Ischemia/reperfusion in Rats

Author: WangXinHe
Tutor: QianYanNing
School: Nanjing Medical University
Course: Anesthesiology
Keywords: Cerebral ischemia Apoptosis Lithium chloride GSK-3β Middle cerebral artery occlusion
CLC: R743.3
Type: Master's thesis
Year: 2007
Downloads: 151
Quote: 0
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Background: Recent studies have found that lithium has a protective effect on nerve cells, but its mechanism is not very clear. Our previous study found that the neuroprotective effects of lithium and its possible reduction of p53 protein and increased bcl-2, HSP70 proteins and activation of Akt related. Akt activation inhibit GSK-3β, and the role of GSK-3β promote apoptosis, thus inhibiting the activity of GSK-3β may be neuroprotective effect of lithium, an important part of the mechanism. In vitro studies have found that lithium inhibits GSK-3β activity, but in vivo studies of lithium relationship with GSK-3β rarely reported. Objective: In the middle cerebral artery occlusion and reperfusion (MCAO / R) after the application of lithium chloride, (a) rat transient focal cerebral ischemia and reperfusion in hippocampal CA1 region of neuronal apoptosis and GSK-3β content and activity changes; (2) the effects of different doses of lithium chloride on transient focal cerebral ischemia and reperfusion in rat hippocampal CA1 neuronal apoptosis and GSK-3β content and activity. MATERIALS AND METHODS: Using nylon thread cerebral artery embolism caused by middle cerebral artery occlusion (MCAO), 90 分钟 suture removal reperfusion prepare short focal cerebral ischemia and reperfusion model. 126 SD rats were randomly divided into normal control group (NO group, n = 6), sham group (SH group, n = 24), ischemia-reperfusion group (IR group, n = 24), lithium chloride 1mmol / Kg group (Lil group, n = 24), lithium chloride 2mmol/Kg group (Li2 group, n = 24), lithium chloride 3mmol/Kg group (Li3 group, n = 24), in addition to the normal control group (NO group) 6 rats, the rest of the group at different time points (MCAO6 hours, MCAO1 days, MCAO3 days, MCAO7 days), each group was randomly divided into four subgroups, each subgroup of six rats. Normal control group (NO group) no treatment, each group of lithium chloride immediately after reperfusion (ie, 90 minutes after MCAO), respectively, corresponding to the dose given intraperitoneal injection of lithium chloride, sham group and ischemia-reperfusion group with an equal volume of saline by intraperitoneal injection of lithium chloride was replaced after every 24 hours once, until death. Hoechst33258 dye staining apoptotic neuronal apoptosis in hippocampal CA1 region changes, indirect immunofluorescence staining hippocampal CA1 GSK-3β content and distribution changes in indirect immunofluorescence staining hippocampal CA1 inactivation of GSK-3β Form p-GSK-3β (Ser9) content and distribution changes. All data were expressed as mean ± standard deviation ((?) ± s) said to Stata8.0 statistical software for univariate analysis of variance, P <0.05 was considered statistically significant. Results: (1) Hoechst33258 staining: NO SH groups and subgroups of rat hippocampal CA1 no right or occasional apoptotic cells, no significant difference between the two groups. SH groups with the same point in time as compared to ischemia-reperfusion group (IR) showed obvious apoptosis (P <0.01), the peak of apoptosis occurs in MCAO1 days (with MCAO6 hours, MCAO3 days, MCAO7 days compared P <0.01). With IR group compared to the same point in time, the number of apoptotic cells lithium chloride group was significantly reduced (P <0.01), but more than SH group (P <0.01). (2) GSK-3β immunofluorescence staining: each rat hippocampal CA1 GSK-3β positive reaction particles was no significant difference between the number (P> 0.05). Normal group (NO group) and sham operation group (SH group) GSK-3β positive reaction particles mainly in the cytoplasm, nucleus rarely; IR lithium chloride group and each group has a large number of GSK-3β distributed in the nucleus, the same point in time lithium chloride IR group and between groups of GSK-3β in the intracellular distribution of no significant difference. (3) p-GSK-3β (Ser9) Immunofluorescence staining: NO group and SH group only in the cytoplasm of a small amount of p-GSK-3β (Ser9). SH groups with the same point in time compared to the IR group p-GSK-3β (Ser9) increased significantly more pronounced increase in the nucleus (P <0.01). The same point in time with IR group after application of lithium chloride, p-GSK-3β (Ser9) increased more significantly (P <0.01), the greater the dose of lithium chloride, p-GSK-3β (Ser9) increased more significantly (P < 0.01 or P <0.05). Conclusion: rat MCAO and reperfusion, hippocampal CA1 neuronal apoptosis, a large number of GSK-3β transferred from the cytoplasm to the nucleus. Lithium chloride dose-dependent reduction of hippocampal CA1 neuronal apoptosis; increased hippocampal CA1 p-GSK-3β (Ser9) expression, inhibition of GSK-3β activity. This may be lithium chloride anti-neuronal apoptosis mechanisms.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Cerebrovascular disease > Acute cerebrovascular disease ( stroke)
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