| Objective: To explore methods to injury time of brain;Using the method of free fall impact to made brain injury model,Comparison ?-APP and PSD95 in the value of brain injury time and degree of brain injury.Methods: According to the location map of brain in rats,Using brain locator locate2.5mm after the rat coronary seam and next to the midline of the skull 2 mm,zone of M2,Using the dental drill to open a 5mm diameter circular window for this point as the center,keep the rat dura intact.From the different height of the set,Using the free fall impact dura mater of the rats,Make different degree of brain injury models,production control as the same time,Namely a circular windows open only the corresponding areas,not impact.After the injury time of 2 h,6 h,12 h,24 h,3 d,7 d,12 d and 14 d,behead method for the death of brain injury model in rats.Take out Brain tissue after using rongeur stripping,Put in 10% neutral formaldehyde fixed liquid 24 h.Then make out to wax afer organize dehydration,paraffin and embedding block,slicing mechanism into tissue section with immunohistochemical technique of target protein staining,Under a microscope photos made into pictures,Then microscopic observation the expression of ?-APP and PSD95 in brain tissue injury and the control group in corresponding brain tissue area.Again with the aid of computer image processing technology to calculate IOD(integral optical density)of ?-APP and PSD95,then analysis of experimental data with the correlation of the degree of brain injury and braininjury form time.According to the regression analysis of the two experiment indicators,distinguish the value of these two indicators in the inference degree of brain injury and brain damage form time.Results: The experimental data using SPSS17.0 software for statistical analysis,1)?-APP analysis results:(1)?-APP IOD value of normal group and 2h,6h,12 h of control group comparison displays P>0.05;IOD value of normal group and 24 h of control group comparison displays P<0.05;IOD value of normal group and 3d,7d,12 d,14d of control group comparison displays P >0.05;The expression of ?-APP in the control group,Within 24 h reaches its peak value after injury,And then gradually decline to normal group level,expression of ?-APP values at different times and in the linear regression quadratic fitting R=0.537;(2)IOD value of normal group and 2h,6h,12 h of the mild injury group comparison displays P>0.05;IOD value of normal group and 24 h,3d of the mild injury group comparison displays P < 0.05;IOD value of normal group and 7d,12 d,14d of the mild injury group comparison displays P>0.05;The expression of ?-APP in the mild injury group,begin to gradually rise in 6h after injury,3days reaches its peak value after injury,And then gradually decline,Basically back to normal group level at the 14 dth after injury,the expression trend like a bell curve,expression of ?-APP values at different times after mild injury and in the linear regression quadratic fitting R=0.320,the linear regression cubic fitting R=0.784;(3)IOD value of normal group and 2h of the moderate injury group comparison displays P>0.05;IOD value of normal group and 6h,12 h,24h,3d of the moderate injury group comparison displays P < 0.05;IOD value of normal group and7 d,12d,14 d of the moderate injury group comparison displays P < 0.05;The expression of ?-APP in the moderate injury group,begin to rise in 2h after injury,3days reaches its peak value after injury,Basically back to slightly higher than the normal group level at the seventh days after injury,the expression trend like mildinjury group,expression of ?-APP values at different times after moderate injury and the linear regression quadratic fitting R=0.361,the linear regression cubic fitting R=0.739(2h-7d);(4)IOD value of normal group and 2h,6h,2h,24 h,3d,7d of the severe injury group comparison displays P?0.05;IOD value of normal group and 12 d,14d of the severe injury group comparison displays P ?0.05;the entire expression trend like mild injury group,the expression trend like mild injury and moderate group;linear regression quadratic fitting R=0.454,the linear regression cubic fitting R=0.820;(5)Comparing different degree of brain injury under the same time:In 2h group,control compared with mild and moderate damage displays P?0.05,control compared with severe damage displays P?0.05;In 6h group,to compare with each other displays P?0.05;In 12 h,24h group,to compare with each other displays P?0.05;control compared moderate severe damage displays P?0.05,Other displays P? 0.05;In 3d group,control compared with the other three groups displays P ?0.05;In 7d group,ontrol compared with mild and moderate damage displays P?0.05;In 7d,12 d group,Comparison between groups displays P?0.05;2)PSD95analysis results:(1)Compare normal group with 2h,6h,24 h,3d,7d,12 d,14d of the control group shows P?0.05,Comparison of normal group and 12 h of the control group shows P?0.05;Expression of PSD95 values at different times and regression quadratic fitting R=0.676;(2)Compare normal group with 6h,12 h,24h,7d of mild injury group shows P?0.05,Compare with other time periods shows P?0.05;The expression of PSD95 in the mild injury group,12 h reaches peak value after injury,and then back to the normal group level,set to rise again,reaches peak value again in the seventh day,back to normal group level again;Expression of PSD95 values at different times periods and regression quadratic fitting R=0.604(2h-3d),the linear regression cubic fitting R=0.998(2h-3d),egression quadratic fitting R=1.000(3d-14d);(3)Compare normal group with 2h,6h,12 h,24h,7d of moderate injury groupshows P?0.05,Compare with other time periods shows P?0.05;the expression trend like mild injury group,reaches peak value again in 12 h and the seventh day,peak is higher than mild group;Expression of PSD95 values at different times periods and regression quadratic fitting R=0.732,the linear regression cubic fitting R=0.963;regression quadratic fitting R=0.814(3d-14d),the linear regression cubic fitting R=0.998(3d-14d);(4)Compare normal group with 2h,6h,12 h,24h,7d,14 d of severe injury group shows P?0.05,Comparison of control group and 12 h of severe injury group shows P?0.05;he expression trend like the same above;Expression of PSD95 values at different times periods and regression quadratic fitting R=0.630(2h-3d),the quadratic regression equation,the linear regression cubic fitting R=0.972(2h-3d);regression quadratic fitting R=0.813(3d-14d),the linear regression cubic fitting R=1.000(3d-14d);(5)Comparing different degree of brain injury under the same time :In 2h group,control compared with mild damage shows P ? 0.05,control compared with moderate and severe damage shows P?0.05;In 6h,12 h,24h group,ontrol compared with each other group shows P ? 0.05;In 3d group,to compare with each other displays P?0.05;In 7d group,ontrol compared with each other group shows P?0.05;In 12 d,14d group,Comparison between groups displays P?0.05.Conclusion: the expression of ?-APP in brain tissue and its damage time there is a certain regression curve relationship,the expression quantity is evident peak value,To infer the brain damage form time to provide some reference.Different degree of brain injury in the same injury time also exist certain differences,Provide certain reference to infer that degree of brain injury.The expression of PSD95 P peaked at 12 h after injury,Three days after injury down to normal group level,7d after injury,peak appeared gain,And 14 d after injury to level slightly higher than the normal group,between expression and injury time have correlation,the inference of brain damage formationtime has a certain value,Different degree of brain injury in the same injury time also exist some differences,Provide a value for refer that degree of brain injury. |
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