The Effect Of Astragalus Injection On Neuron Apoptosis And Expression Of JNK3in Hippocampus Of Cerebral Ischemia Reperfusion Rats

Ischemic cerebrovascular disease is a common and serious ailment inclinic, and reperfusion injury is one of its significant mechanisms, moreover, itbecomes an important factor which restricts curative effect of ischemiccerebrovascular disease, so it is an urgent task in medical profession that probeinto pathophysiological mechanism of cerebral ischemia-reperfusion andenhance prevention and therapy. In the development process of ischemiccerebrovascular disease, apoptosis is a mechanism of neuronal damage, andthe expression of apoptosis-related gene c-Jun N terminal kinase3is relativeto neuron apoptosis. Astragalus injection is commonly used to cure cerebralvessel diseases, which can inhibit cell apoptosis, but by which signaltransduction executes it has not been illustrated. We can probe into themolecular mechanism which astragalus injection protects brain throughinvestigate the effect of astragalus injection on neuron apoptosis andexpression of JNK3after cerebral ischemia reperfusion in hippocampus ofrats.The first partThe effect of different doses of astragalus injection on neuron apoptosisand expression of caspase-3in hippocampus of rats after cerebral ischemiareperfusionAim:To investigate the effect of different doses of astragalus injection onneuron apoptosis and expression of caspase-3in hippocampus of rats aftercerebral ischemia reperfusion. Methods:Select healthy male SD rats randomly divided into3groups: the shamoperation group, model group and astragalus injection group. The astragalusinjection group was then subdivided into5subsets, according to the astragalusinjection doses of2ml/kg,4ml/kg,6ml/kg,8ml/kg and10ml/kg. Thefour-vessel occlusion method is used to built rat model of cerebral ischemiareperfusion. After transient global cerebral ischemia (30min) and reperfusion24h, method of HE staining is used to observe morphological changes inhippocampal CA1zone, TUNEL staining is used to measure the neuroneapoptosis in hippocampus and Western blot is used to measure the expressionof caspase-3protein.Result:HE staining showed that: the neurons in hippocampal CA1zone of rats inmodel group appeared significantly pathological changes, irregulararrangement, coagulation necrosis and obvious deletion. Compared withmodel group, pathological changes of the neurons in astragalus injection4ml/kg,6ml/kg,8ml/kg and10ml/kg group had significant improved, and theeffect of large doses is more obvious. TUNEL staining and western blotting:Compared with the sham operation group, the apoptosis index and expressionof caspase-3protein increased in the model group (p<0.05). Compared withthe model group, the apoptosis index and expression of caspase-3proteindecresed in the astragalus4ml/kg,6ml/kg,8ml/kg and10ml/kg group(p<0.05), but there is no difference in astragalus2ml/kg group (p>0.05).Compared with the4ml/kg group, the apoptosis index and expression ofcaspase-3protein decresed in the astragalus6ml/kg,8ml/kg and10ml/kggroup (p<0.05), but there is no difference between the later three group(p>0.05).Conclusion:1. four-vessel occlusion method can successfully built cerebral ischemiareperfusion rat model, and astragalus injection can improve pathologicalchanges in hippocampal CA1zone of cerebral ischemia reperfusion rats.2. Astragalus injection can decrease neuron apoptosis and expression of caspase-3protein, moreover that relate to the dose of the astragalus injection.All of the5different doses of astragalus injection, the6ml/kg is the best one.The second partThe effect of astragalus injection on the expression of JNK3inhippocampus of cerebral ischemia reperfusion ratsAim:To investigate the effect of astragalus injection on the expression of JNK3in hippocampus of cerebral ischemia reperfusion rats.Methods:Select healthy male SD rats randomly divided into4groups: the shamoperation group, model group, astragalus injection group and originalastragalus injection group. Model group, astragalus injection group andoriginal astragalus injection group after transient global cerebral ischemia(30min), was then subdivided into7subsets, according to reperfusion times of0h,0.5h,2h,6h,24h,72h and120h. Methods of immunohistochemistry andWestern blot were used to measure the expression of JNK3protein, real-timePCR were used to measure the expression of JNK3mRNA.Results:Compared with the sham operation group, the expression of JNK3proteinand mRNA in hippocampus very time points increased obviously in modelgroup except120h (p<0.05); compared with the model group, the expressionof JNK3protein and mRNA in hippocampus very time points decreasedobviously in astragalus injection group besides120h (p<0.05), however,theexpression of JNK3protein and mRNA in hippocampus very time points hadno obvious change in original astragalus injection group (p>0.05).Conclusion:1. The expression of JNK3protein and mRNA increased obviously inrat’s hippocampal neurons after cerebral ischemia reperfusion, this accordedwith hippocampal neurons apoptosis. It is indicate that JNK3probablyparticipate in rat hippocampal neurons injury after cerebral ischemia reperfusion.2. Astragalus injection can inhibit the expression of JNK3mRNA inhippocampus of cerebral ischemia and reperfusion rats, moreover, it caninhibit the expression of JNK3protein, accordingly inhibits hippocampalneuron apoptosis.Conclusions1. Astragalus injection can decrease neuron apoptosis and expression ofcaspase-3protein, moreover that related to the dose of the astragalus injection.All of the5different doses of astragalus injection, the6ml/kg is the best one.2. Astragalus injection can inhibit the expression of JNK3mRNA andprotein in hippocampus of cerebral ischemia and reperfusion rats, accordinglyinhibits hippocampal neuron apoptosis.