The Effect Of Inhibition Of Pten Nuclear Translocationin In Hypoxic Ischemic Brain Damage

Part1HIBD animal model and the level of PTENin nuclearObjective: In order to establish the newborn rats hypoxic ischemicbrain damage (HIBD) model, newborn Sprague-Dawley (SD) rats wereused. Whether the tumor suppressor of PTEN in nucleus increased afterhypoxia ischemia was observed.Methods:7-day newborn SD rats were randomly divided into HIBDgroup and control group. SD rats were fixed on the operation panel afterinhalation anesthetic isoflurane, the neck skin was disinfected whit75%alcohol, the left common carotid artery was separated and ligated at theproximal and telecentric parts, then blood vessels were cut off.After cut offthe left common carotid artery ligation, closing the small wound.Take backon their mother side, rest two hour, then HIBD group into the airtight anoxicbox,filling with8%oxygen and92%nitrogen mixture, last2.5hours, takeback litters on the female side.Control group no hypoxic ischemic.Respectively in3h,6h,12h,24h,3d,7d after ligation of blood vessels,separate left and right side of the hippocampus and cortex on the ice, Thenextracting protein to do the western blot experiment, In order to detect thelevels of pten in the nucleoprotein. Results: After hypoxic ischemic brain damage in3h,6h,12h, thelevel of pten in nuclear is high, after24h HIBD group were almost returnedto control levels.Conclusion: When Hypoxia ischemia, the level of pten in nuclearcome from increasing to normal.Learn about the time course is helpful for usto find the best window of treatment time. Part2The polypeptide Tat-K13treatmentof HIBD and animal behavior testObjective: According to the first part of result, detecting the doseeffect relationship of peptide Tat-K13and observe the rats behaviorchange after using the peptide.Method: First of all, inject different doses of Tat-K13peptide (0,2.5,5,10and20mg/kg, i.p.) or Tat-K13R control peptide, give a medicinebefore the hypoxia, and hypoxia3h, take brain tissue after hypoxia12h,carry on the left side of the hippocampus and extracte Nucleoprotein，western blot detect the level of PTEN in nuclear. When dose effectrelationship stability on animal models, respectively, give a medicine athypoxia3h,24h,48h. when4to6weeks,observe the behavior changes.Results: According to the dose effect relationship, determine the finaldosage of10mg/kg, the weight in the newborn rats after HIBD, no statisticaldifferences in the model. High plus maze test, no statistical differences to tellus their emotional memory function have no damage, turn rods and holdingpower experiments detect its movement ability, statistically significant,water maze, avoid dark evading experiments test its spatial memory ability,statistically significant. These behavioral experiment data for our prompt is:Hypoxic brain injury can cause movement and cognitive damage, inhibitpten in nuclear can protect neurons.Conclusion: Inhibition of PTEN nuclear transfer have protective effecton hypoxic ischemic brain damage.