Activation Of The Death Receptor Apoptosis Pathway Which Fas Molecule Started And Intervention Of HWTX-I In Hippocampus Of Global Cerebral Ischemia Reperfusion Rats

Objective:To observe the effects of HWTX-I on morphological changes of nissl staining hippocampus CA1 pyramidal neurons of global cerebral ischemia reperfusion rats by optical microscope, and on morphological changes of mitochondria of the hippocampus CA1 pyramidal neurons by electron microscope, and on mRNA and protein expressions of Fas apoptosis pathway correlation factors, by RT-PCR and Immunohistochemical Assay, respectively, in order to investigate the correlation of HWTX-I and the dead signal transduction pathway which started by Fas molecule. To study the effectiveness and possible molecular mechanism of HWTX-I to avoid hippocampus pyramidal neurons from cerebral ischemic reperfusion injury, to aim to provide academic and experimental evidences for developing a new neuroprotective drug.Methods:48 rats were randomly divided into three groups: sham-operated group, nonadministered group and administered group, with 16 rats in each group. 8 rats of each group were used to observe nissl staining morphological changes of hippocampus CA1 pyramidal neurons of global cerebral ischemia reperfusion rats by optical microscope, and morphological changes of mitochondria of the hippocampus CA1 pyramidal neurons by electron microscope ,and protein expressions of Fas apoptosis correlation factors by Immunohistochemical Assay. The other 8 rats of each group were used to observe mRNA expression of Fas apoptosis pathway correlation factors by RT-PCR. All rats (except for sham-operation group)were used to construct "4-vessel occlusion" global cerebral ischemia reperfusion injury model combined with tube placement in subarachnoid space after proper improvement.Results:1. By nissl staining, multilayer pyramidal neurons that ranged orderly and densely were clearly observed, in sham-operated group; However pyramidal neurons, in nonadministered group, ranged confusedly and sparsely with incomplete layers;In administered group, arrangement of multilayer pyramidal neurons sparse but orderly distributing were observed.2. By electron microscope, mitochondrial cristae that were clear, without breakage, were observed, in sham-operated group;In nonadministered group, mitochondrial cristae were unclear, and vacuolization occurred;In administered group,clear mitochondrial cristae without obvious breakage, were observed.3. There were almost no protein expressions of Fas apoptosis pathway correlation factors, in sham-operated group;In nonadministered group, stronger protein expressions of Fas apoptosis pathway correlation factors and morepositive cells occurred, than in other groups. In administered group, positive cells were sparse.4. The statistical results indicated that average positive unit values of Fas, FasL, FADD and Caspase-3 in nonadministered group were the highest of three groups, followed by administered group and sham-operated group respectively;The protein expressions of Fas in nonadministered group and administered group were very significantly different from that in sham-operated group (P＜0.01),respectively, and there were significant differences between nonadministered group and administered group (P