| Objective: applying focal ischemia-reperfusion model of SD rat,to observe the change of expression of Interleukin-1β(IL-1β)and transforming growth factor-β1(TGF-β1)and the nerve growth factor on every time rats after focal cerebral ischemical reperfusion injury in Sprague-Dawley(SD)rat,to investigate the brain protecting mechanism of nerve growth factor and to offer the theory of curing ischemic cerebrovascular disease.Method: 55 healthy adult SD rats, female and male unlimited,the weight 200-250g,sweep class ,feed environment temperature 21℃,take a meal water freely.They were randomly divided into a false surgical operation group (n=5)(Group1);Matched group(n=30),according to ischemic 2h reperfusion 2h,6h,12h,24h,48h,72h is divided into 6 sets(respectively is 2,3,4,5,6 and 7 groups ),each 5;The nerve growth factor group (n=10)(Group 8) and matched group (n=10)(Group 9)in physiology brine. According to Nagasawa report, to anaesthetize animal with 10% chloral hydrate(0.35ml/100g belly cavitieses inject),take to lie on the back one fixed, go right side MCAO surgical operation in the surgical operation on the stage, the neck slices exact center ,separating a right side common carotid artery, external carotid artery and internal carotid artery,ligature the carotid artery near heart and external carotid 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 implement reperfusion. After the animal revivals, observe its posture and behavior, judge the MCAO model to be successful. After the false surgery group inserts the nylon line to extract immediately, old step with surgery group. After the ischemia reperfusion model animals to be sober, the left forelimb flexure adduction rat is the research object. The 8th group of rats after reperfusion 2h and 24h were intramuscular injection NGF (NGF 500BU/0.1ml/100g), the 9th group separately pours into the same volume by the similar method the aseptic physiological saline. Separately observes several groups of animals following targets: Zea Long 5 mark system standard grades carries on each group of rats nerve function appraisal; to examine group TGF-β1,IL-1βexpression after the HE dyeing and the immunity histochemistry in the rats brain.Results: (1) the nerve function grades:2-9 group grade are2.000±0.7071,2.200±0.4472,2.400±0.5477,2.600±0.5477,2.800±0.4472,3.000±0.0000,1.400±0.6253,2.300±0.3650,2.200±0.6903,3.100±0.0038,The nerve growth factor group (8th group )with correspondingly to ischemia- reperfusion of the salt water(9th group )to compare, the nerve function barrier obviously reduces.(P<0.01).(2)After the ischemia-reperfusion, in the big mouse brain the IL-1βexpression quantity increase, and reperfuse to 2 hours to reach the peak again, later will gradually decrease.TGF-β1 reaches the peak in 24h groups, later will reduce. (P<0.01). With ischemia- reperfusion, to compare, in their respectively peak time, the nerve growth factor (intervention),may cause in the brain IL-1βdecrease and TGF-β1 expression increase.(p<0.01)Conclusion: The intramuscular injection nerve growth factor has the obvious protective function to ischemia-reperfusion rats, IL-1βdecreases and TGF-β1increases possibly is one of its mechanisms. |
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