Effect Of Nerve Growth Factor On Expression Of P-PKB And Fas-L Gene Protein In Rat After Cerebral Ischemial Reperfusion

Objective:Adopting focal cerebral ischemia and reperfusion model of Sprague-Dawley rat ( SD rat ) to observe injury of the neural function,expression of p-PKB(phosphoryla -tion of Protein Kinase B) and Fas-L gene protein on every time after reperfusion of local cerebral ischemia and to observe the protective function of NGF in the brain of rat and further to investigate the brain protecting mechanism of NGF and to offer the theoretical basis for clinical therapy of ischemia cerebrovascular disease.Method:55 healthy adult SD rats unlimited sex take the meal and water freely and at the same time they must be provided with follow condition: their weight controled between 200-250g, cleanly class, environment temperature of 21 centigrade degree.They were randonly divided into false surgical operation group(n=5)(Group 1),matched groups(n=30) which according to ischemic time is divided into 2h group,6h group,12h group,24h group,48h group,72h group(5 rats in each group) ,the NGF groups which includ 8f group (after reperfusing 6h injecting NGF)and 8p group (after reperfusing 72h injecting NGF) (5 SD rats in each group), saline groups which include 9f group(after reperfusing 6h injecting saline) and 9p group(after reperfusing 72h injecting saline)(respectively 5 SD rats). According to Nagasawa report, to anaesthetize animal wuth 10% chloral hydrate(0.35ml/100g belly cavitieses inject),take to lie on the back on the stage,the neck slices exact center,separating a right side common carotid artery near heart and external caroid artery branch start,slice to shear 1 in the carotid artery branch start,head carry to dip to have a pure paint of diameter for 0.20mm fishing thread carries to insert toward internal carotid artery,continuing to push forward until feel slight resistance,then about 18 millimeters,hint to have already got to brain in middle cerebral artery start department.After 2 hours slowly promote the fishing thread then to inplement reperfusion.After the animal revivals observes its posture and the behavior to judge the MCAO modle weather it is successful.After the false sergery group inserts the nylon line to extract immediately and other steps are same with surgery group.After the ischemia-reperfusion model animals to be sober,the right forelimb flexure adduction rat is the research object.The 8th group of rats after reperfusing 5 minutes were intramuscular injection NGF(500BU/0.1ml/100g),the 9th groups seperately pour into the same volume of saline in the same way of 8th group .At last ,observing the following targets of several groups animals:zea long 5 mark system standard grades is carried to each group of rats for nerve function appraisal and examining the expression of Fas-L and p-PKB protein after the HE dyeing and the immunity histochemistry in the rats brain.Result:1,The nerve function grade:2-9group grade are2.000±0.6032,2.200±0.4112,2.400±0.6880,2.600±0.3348,2.800±0.4621,3.000±0.5864, 1.400±0.6253(8f),2.300±0.3650(8p),2.200±0.6903(9f),3.100±0.0038(9p),.Being respectively compared with normal saline group:9f and 9p group ,disfunction of NGF group(8f group and 8p group are obviously alleviated.(p<0.01)2,In the SD rats'brain the p-PKB protein has a low-level expression normally and increased after ischemia-reperfusion,peaked at 72h after reperfusion.The NGFcan increase the expression of the p-PKB protein at 72h.(p<0.01). Fas-L protein has a low-level expression normally and increased after ischemia reperfusion,peaked at 6h after reperfusion.The NGFcan reduce the expression of the Fas-L protein at 6h.(p<0.01).Results above show that NGF being given in the ischemia early time can increase the expression of p-PKB protein reduce the expression of Fas-L protein.(p<0.01).Conclusion:The NGF in the way of intramuscular injection has obviously protective function for ischemia-reperfusion Rat. Increasing p-PKB and decreasing Fas-L gene protein may be one of its mechanisms.