Effect Of Plasminogen Activator On Brain Tissue In Rats

ObjectiveThe normal brain tissue of Wistar rats was used as the experimental basis,and the usual doses of uPA and rtPA were given to the caudate nucleus for injection.The morphological changes of the brain were observed after 2 hours,4 hours,and 6 hours respectively.The brain tissue edema,blood-brain barrier permeability,and neuronal cell apoptosis were detected.The purpose of this study was to investigate the safety of normal brain tissue using the doses of uPA and rtPA commonly used clinically,and to search for the least impact on normal brain tissue.Dosage and dissolution time.MethodsThis experiment used 108 SPF Wistar rats,adult males,weighing 350 to 400 g.Wistar rats were randomized into groups: uPA group(administered group: I),rtPA group(administered group: II),and the concentration of uPA administered was set at 30,000 U/ml,50,000 U/ml,and 100,000 U/ml.The concentration of ml and rtPA was set to 0.8 mg/ml,1.5 mg/ml,and 2 mg/ml,and each dose group was divided into 2 h group,4 h group,and 6 h group,with 6 mice in each group.Rats in the administration group were injected with 20 ul of uPA or rtPA in the caudate nucleus.The brain tissue was removed and paraffin sections were taken.HE staining was used to observe the effects of fibrinolytic drugs on neuromorphology in rats.Immunohistochemistry was used to detect brain tissue aquaporin-4(AQP-4)and matrix metalloprotein-9(MMP-9)Expression of B-lymphoma-2(Bcl-2)and caspase-3 protein.Content and results1.Effect of Plasminogen Activator on Morphology of Rat BrainBy comparative observation,uPA administration group had less bleeding and more tissue edema,but the degree of edema was significantly improved in the uPA at 550,000 U/ml and 6 hours.Inflammatory cells were infiltrated but not obvious.The bleeding was more common in the rtPA group than in the normal saline group.Although there was less bleeding than the normal saline group,the area of tissue edema was small,but the loosening of tissues was more common.In the rtPA 1.5mg/ml,2h group,there was no hemorrhage and edema in the brain tissue.Light,neuronal cells pyknosis,degeneration than other rtPA administration group.2.Effect of plasminogen activator on the expression of AQP4 and MMP9 protein in rat brainThe expression level of AQP-4 in the brain of uPA-administered rats was analyzed:(1)uPA had different doses,uPA 3 million U/ml and uPA 5 million U/ml were statistically different(P<0.05);(2)uPA had different effects at the same time,the expression of AQP-4 was the same.(3)The interaction between them(P<0.05)was different at different doses and different time of action.The expression of uPA5 million U/ml6hAQP-4 was the lowest,and the expression of uPA3 million U/ml6hAQP-4 was the highest.The expression level of AQP-4 in the brain of rtPA-administered group was analyzed:(1)The different doses of rtPA,the expression of AQP-4 was the same;(2)The different time of rtPA,the expression of AQP-4 was the same;(3)The different dosing concentration and the interaction between them was different(P<0.05),the expression of rtPA2mg/ ml2hAQP-4 was the lowest,and the expression of rtPA2mg/ml6hAQP-4 was the highest.The expression level of MMP-9 in the brain of uPA-administrated rats was analyzed:(1)uPA had different doses,uPA 3 million U/ml and uPA 5 million U/ml were statisticallydifferent(P<0.05);(2)uPA had different effects the time,between 2h and 4h,was statisti cally significant(P<0.05),there was a statistical difference between 4h and 6h(P<0.05);(3)The interaction between different dosing concentrations and different action times(P< 0.05)The expression of MMP-9 was the lowest in uPA at 550,000 U/ml,while the expression of MMP-9 was highest at uPA310,000 U/ml at 6 h.The expression level of MMP-9 in the brain of rats treated with rtPA was analyzed:(1)There was a statistical difference in the concentration of rtPA 0.8mg/ml and rtPA 1.5mg/ml(P<0.05),rtPA 0.8mg/ml Compared with rtPA2mg/ml,there was statistical difference(P<0.05);(2)rtPA had different time of action,MMP-9 expression was the same;(3)The interaction between different doses and different action time(P<0.05),rtPA2 mg/The expression of MMP-9 was lowest in ml6 h,and the expression of MMP-9 was highest in rtPA 1.5mg/ml6 h.3.Effect of plasminogen activator on cell apoptosis in rat brainThe expression of Bcl-2 in the brain of uPA-treated rats was analyzed.The concentration of 1uPA at different doses was the same as that of Bcl-2.There was a statistically significant difference between 2h and 4h at 2uPA(P<0.05).There was a statistical difference between 6h(P<0.05);3The interaction between different doses and different time of action(P<0.05),uPA100000U/ml2hBcl-2 expression was the lowest,uPA510000U/ml6hBcl-2 The highest expression.The expression of Bcl-2 in the brain of rats treated with rtPA was analyzed: 1 There was a statistical difference between rtPA 0.8mg/ml and rtPA 1.5mg/ml(P<0.05),rtPA 1.5mg/ml There was a statistically significant difference between rtPA and 2mg/ml(P<0.05),there was a statistical difference between rtPA 0.8mg/ml and rtPA 2mg/ml(P<0.05);2 rtPA had different time of action,statistically between 2h and 4h.Differences(P<0.05),statistical differences between 4h and 6h(P<0.05),statistical differences between 2h and 6h(P<0.05);3 between different dosing concentrations and different action times The interaction,rtPA1.5mg/ml4hBcl-2 expression was the lowest,rtPA2mg/ ml2hBcl-2 expression was the highest.The expression level of Caspase-3 in the brain of uPA-administered rats was analyzed.There were statistical differences between 1 uPA and different doses of uPA,uPA 3 million U/ml and uPA 5 million U/ml(P<0.05).uPA 3 million U/ml There was a statistically significant difference between uPA and 10 million U/ml(P<0.05);2 uPA had different time of action,and the expression of caspase-3 was the same;3 the interaction between different doses and time of action(P<0.05),uPA3 Million U/ml6hCaspase-3 expression was the lowest,and uPA10 million U/ml6hCaspase-3 was the highest.The expression level of Caspase-3 in the brain of rats treated with rtPA was analyzed: 1 There was statistical difference between rtPA 0.8mg/ml and rtPA 1.5mg/ml(P<0.05),rtPA 0.8mg/ml There was a statistical difference between rtPA and 2mg/ml(P<0.05);there was a statistically significant difference between 2h and 4h in 2rtPA(P<0.05),and there was a statistical difference between 4h and 6h(P<0.05).;3 The interactions between different doses and different time of action(P<0.05),rtPA1.5mg/ml4hCaspase-3 expression was the lowest,rtPA0.8mg/ml2 h expression was the highest expression of caspase-3.4.Comprehensive analysis of immunohistochemical resultsAnalysis of four indicators of comprehensive immunohistochemistry,uPA 550,000 U/ml uPA group,the smallest impact on brain tissue;rtPA administration group,rtPA2mg/ ml group on the brain tissue minimal impact;the same effect time 4h,uPA5 The concentration of two drugs,Wan U/ml and rtPA 2 mg/ml,had the least influence on the brain tissue.The results of immunohistochemical analysis of uPA5 million U/ml4 h group and rtPA2mg/ml4 h group showed that the expression of AQP-4 in the uPA 5 million U/ml 4h group was approximately the same as that in the rtPA 2mg/ml 4h group,but the expression of MMP9 and Caspase-3 in the uPA 5 million U/ml 4h group was significant.Lower than rtPA2mg/ml4 h group,Bcl-2 expression was significantly higher than rtPA2mg/ml4 h group,so uPA510000U/ml4 h had minimal impact on brain tissue.Conclusion1.In the uPA-administered group,the uPA 5 million U/ml group had the least influence on the brain tissue;in the rtPA-administered group,the rtPA 2 mg/ml group had the smallest influence on the brain tissue.2.At the same action time of 4 h,the concentration of two drugs,uPA 50 000 U/ml and rtPA 2 mg/ml,had the least influence on the brain tissue.3.Compared with rtPA2mg/ml4 h,uPA550,000 U/ml4 h had little influence on brain tissue,and the drug had a low price and had a higher promotion value.