| Objective: Observing the effects of huwentoxin -I(HWTX-I) on activity of SOD, CAT in brain and the content change of MDA, on morphological change by Nissl staining of hippocampus CA1 pyramidal neurons of global cerebral ischemia reperfusion rats, on the mitochondria morphological change in the hippocampus neurons, on the expression of apoptosis correlation factor Bax, bcl-2 mRNA and protein, in order to research HWTX-I's mechanism of relieving global cerebral ischemia reperfusion damage , so as to offer new effective treatment to cranicerebral injury in clinic.Methods: 88 grown-up male SD rats, which were randomly divided into 5 groups: the sham-operation group(n=24), the physiological saline group(n=24), the HWTX-I 0.5μg/kg group(n=8), the HWTX-I 1.0ug/kg group(n=24),the HWTX-I 2.0μg/kg group(n=8). 8 rats of each group were detected activity of SOD, CAT in brain and the content change of MDA, 8 rats of the sham-operation group , the physiological saline group , the HWTX-I 1.0μg/kg group were used to observe morphological change by Nissl staining of hippocampus CA1 pyramidal neurons of global cerebral ischemia reperfusion rats and the expression of apoptosis correlation factor Bax, bcl-2 protein, other 8 rats of the sham-operation group, the physiological saline group, the HWTX-I 1.0μg/kg group were used to the mitochondria morphological change in the hippocampus neurons and the expression of apoptosis correlation factor Bax, bcl-2 mRNA. All rats (except for the sham-operation group) were used in the global cerebral ischemia model combined with the method to playing anaesthesic tube into intrathecal space on the study. Results:(1)1. compared with the physiological saline group, the brain homogenate's activity of SOD of three administration groups risen with the rising of dose, the difference have no remarkable statisticalsignificance. 2. compared with the physiological saline group, the brain homogenate's content of MDA of three administration groups declined with the rising of dose, the difference have remarkable statistical significance between the physiological saline group and 2.0ug/kg group(P<0.05). 3. compared with the physiological saline group, the brain homogenate's activity of CAT of three administration groups risen with the rising of dose, the difference have remarkable statistical significance between the physiological saline group and 2.0ug/kg group(P<0.05). (2) by Nissl staining the sham-operation group hippocampus CA1 pyramidal neurons' several layers pyramidal neurons has been observed, their arrangement is tidy and tightness;the HWTX-I l.Oug/kg group's hippocampus pyramidal neurons sparsely distributed slightly;the physiological saline group's pyramidal neurons' arrangement is loose and disorder, hierarchy have no integrity, pyramidal neurons are distributed sparsely.(3) by electron microscope the sham-operation group hippocampus neurons' mitochondrial crista is clear to observe, the breaking of mitochondrial crista aren't seen;the HWTX-I l.Oug/kg group hippocampus neurons' breaking of mitochondrial crista aren't obvious;the physiological saline group hippocampus neurons' the mitochondrial crista are difficult to observe, the cavity are easy to observe.(4) By immunohistochemistry staining the sham-operation group's expression of Bax positive neurons have not almost been seen in hippocampus CA1 subfield;the HWTX-I l.Oug/kg group's the expression of Bax positive neurons have been seen in hippocampus CA1 subfield. the positive neurons are distributed sparsely;the physiological saline group's expression of Bax positive neurons have been seen in hippocampus CA1 subfield, positive neurons are distributed densely.(5) By immunohistochemistry staining the sham-operation group's expression of a few bcl-2 positive neurons have been seen in hippocampus CA1 subfield. the physiological saline group: theexpression of positive neurons have been seen in hippocampus CA1 subfield, the positive neurons are distributed sparsely, the HWTX-I l.Oug/kg group: the expression of positive neurons have been seen in hippocampus CA1 subfield, positive neurons are distributed densely.(6) The expression of Bax mRNA in the rats' hippocampus of HWTX-I l.Oug/kg group is less than the expression in the physiological saline group by RT-PCR. Although the difference have no remarkable statistical significance between the HWTX-I 1 .Oug/kg group and the physiological saline group, the expression of Bax mRNA in the HWTX-I l.Oug/kg group appear downtrend than the physiological saline group.(7) The expression of bcl-2 mRNA in the rats' hippocampus of HWTX-I l.Oug/kg group is more than the expression in the physiological saline group. Although the difference have no remarkable statistical significance between the HWTX-I 1 .Oug/kg group and the physiological saline group, the expression of bcl-2 mRNA in the HWTX-I 1 .Oug/kg group appear upward trend than the physiological saline group.Conclusion:1. The administration of HWTX-I in intrathecal space can obviously relieve the damage of free radical in global cerebral ischemic rats' brain homogenate.2. The administration of HWTX-I in intrathecal space can maintain mitochondria normal configuration of hippocampus neurons in the global cerebral ischemic reperfusion rats, stabilize the hippocampus CA1 subfield pyramidal neurons' distribution, protected hippocampus pyramidal neurons in rats.3. The administration of HWTX-I in intrathecal space decreased the expression of apoptosis factor-Bax mRNA and protein in the hippocampus of global cerebral ischemic reperfusion rats, increase the expression of anti-apoptosis factor—bcl-2 mRNA and protein in the hippocampus of global cerebral ischemic reperfusion rats, has obviously neuroprotective effects on rats hippocampus neuronsdamage induced by global cerebral ischemic reperfusion.
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