| Part Ⅰ Expression and interaction of long-term plasticity-related protein family Plpprs,mitochondrial function-related proteins and zinc ion transporter molecules in penicillin-induced recurrent seizuresObject:To investigate the long-term development,neurobehavioral,cognition and hippocampal morphological damage induced by penicillin in rats with hippocampus and cerebral cortex plasticity-related protein family Plpprs,mitochondrial function-related protein molecules and zinc ion transporter molecule expression changes and interactions.Method:200 male SD(day 21 days,P21)rats were randomly divided into 2 groups:Control group(n=80),modeling group(n=120).Rats in the modeling group were established with penicillin-induced developmental epilepsy model:penicillin was injected intraperitoneally in each rat at a dose of 5 million units/kg,once every other day,and a total of 6 injections were performed(P21-31).According to the inclusion criteria,80 RS groups(n=80)were obtained.The control group rats were intraperitoneally injected with the same dose of saline at the same time as the RS group.Rat body weight was detected every other day.After modeling for 7 days,14 days,21 days,and 28 days,detect the development of neuroreflex,emotion,learning and memory index changes;Neo-Timm’s staining to observe hippocampal mossy fiber germination and amygdala Timm staining;Nissl staining to observe number and morphology of hippocampus and amygdala neurons;Golgi staining to observe neuronal axon length,grading and axon number;Real-Time RT-PCR detection to observe mRNA expression of hippocampal plasticity related protein family Plpprs,zinc ion channel protein ZnTs and mitochondria-related protein;Western-Blot was used to detect the expression of plasticity-related protein families Plpprs and mitochondria-associated proteins in the hippocampus and cerebral cortex.Results:(1)Body weight:There was no statistical difference in weight between the Control group and the RS group at each time point on the first day of modeling.At 7th day(P38,n=20),14th day(P45,n=20)and 21th day(P52,n=20)point after the establishment of penicillin-induced development repeated seizure brain injury model,The weight of the RS group was significantly lower than that of the Control group at the second injection of penicillin(P<0.05).At the 28th day(P59,n=20)time point,the body weight of the RS group at the third injection of penicillin was significantly lower than that of the Control group(P<0.05).When the RS group begins to appear in this low-weight state,it will continue until the day of execution.At 7th day(P38,n=20),14th day(P45,n=20)and 28th day(P52,n=20)point,the average daily weight gain rate of the RS group was significantly lower than that of the Control group(P<0.05).The average daily weight gain rate of the RS group after 21 days of the last penicillin injection was lower than that of the Control group,but it was not statistically significant;(2)Neurodevelopment:In the forearm suspension test,the forebrain suspension reflex time in the RS group at 7 days,14 days,21 days,and 28 days was significantly lower than that in the Control group,and the difference was statistically significant(P<0.05).In the negative geotaxis test,the negative geotaxis reflex time of the RS group at 7 days,14 days,21 days,and 28 days was significantly higher than that of the Control group,and the difference was statistically significant(P<0.05).In the cliff avoidance test,the cliff avoidance reflex time in the RS group at 7 days,21 days,and 28 days was significantly higher than that in the Control group,the difference was statistically significant(P<0.05),and the RS group was at 14 days.The cliff avoidance reflex time was higher than the Control group,but the difference was not statistically significant.In the plane righting test,the plane righting reflection time of the RS group was significantly higher than that of the Control group at the 21-day time point,and the difference was statistically significant(P<0.05).The difference between the other three time points was not statistically significant;(3)Neurobehavioral:In the open field experiment,the number of horizontal crossing grids in the RS group at 7 days,14 days,21 days,and 28 days was significantly lower than that in the Control group,and the difference was statistically significant(P<0.05).The number of beards in RS group was significantly higher than that in Control group at 7 days,21 days and 28 days.The difference was statistically significant(P<0.05).The number of beards in RS group was higher than Control group at 14 days,but the difference was not statistically significant.The number of standing of the hind limbs in the RS group at 7 days,14 days,21 days,and 28 days was significantly lower than that of the Control group,and the difference was statistically significant(P<0.05).(4)Learning and memory:In the new object recognition experiment,the cognitive index of RS group at 14 days,21 days and 28 days was significantly lower than that of Control group,the difference was statistically significant(P<0.05);the cognitive index of the RS group at 7 days was lower than that of the Control group,but the difference was not statistically significant;(5)Neo-Timm’s staining:1.Hippocampus:The number of Timm particles stained in the CA3 and DG areas of the RS group at 21 days and 28 days was significantly higher than that of the Control group,the difference was statistically significant(P<0.05);There were no significant differences between the RS group and the Control group in the CA3 and DG areas at 7 and 14 days.2.amygdala:The coloration density of the amygdala in the RS group was significantly higher than that in the Control group at 21 days and 28 days,and the difference was statistically significant(P<0.05).There was no statistically significant difference in the color density of the amygdala between the RS group and the Control group at 7 and 14 days.(6)Nissl staining:In the RS group,the number of neurons in the hippocampal CA1 area,CA3 area and amygdala area at 7 days,14 days,21 days and 28 days was significantly lower than that of the Control group(P<0.05).The structure of the element is disordered,and the cytoplasmic Nissl staining is uneven.(7)Real-Time RT-PCR:①.plasticity-related protein family Plpprs:Penicillin treatment significantly increased the expression of Plppr3,Plppr4 and Plppr5 mRNA in SD rats at 28 days compared with Control group,the difference was statistically significant(P<0.05);the expression of Plppr3,Plppr4,and Plppr5 mRNA in RS group at other time points was not statistically significant compared with Control group.There were no statistical differences in the expression of Plppr1 and Plppr2 mRNA of the RS group at 7 days,14 days,21 days,and 28 days compared with the Control group.②.ZnTs gene:Penicillin treatment significantly increased the expression of ZnTl mRNA in the RS group at 28 days compared with the Control group,the difference was statistically significant(P<0.05).The expression of ZnT1 mRNA in the RS group was higher than that in the Control group at 14 days,but it was not statistically significant.There was no statistical difference in ZnT1 mRNA expression between the RS group and the control group at 7 days and 21 days.The expression of ZnT3 mRNA in RS group was significantly higher than that in Control group at 21 days and 28 days,the difference was statistically significant(P<0.05).There was no statistical difference in ZnT3 mRNA expression between the RS group and the Control group at 7 days and 14 days.There was no statistical difference between ZnT6 and ZnT7 mRNA expression in RS group at 7 days,14 days,21 days,and 28 days.③.Mitochondrial function-related gene:Penicillin treatment significantly increased the expression of Bnip3L and BAD mRNA in SD rats at 28 days compared with Control group,the difference was statistically significant(P<0.05).The expression of Bnip3L and BAD mRNA in RS group at 7 days,14 days and 21 days was higher than that in Control group,but it was not statistically significant.The expression of hypoxia-inducible factor HIF-1α and mTOR mRNA in RS group was significantly higher than that in Control group at 14 and 28 days,the difference was statistically significant(P<0.05).The expression of hypoxia-inducible factor HIF-la and mTOR mRNA in RS group was higher than that in Control group at 7 and 21 days,but it was not statistically significant.The expressions of HDAC1,FOUNDC1 and PARKIN mRNA in RS group were significantly higher than those in Control group at 7 days,14 days,21 days and 28 days,and the difference was statistically significant(P<0.05).The expression of sesn2 mRNA in RS group was significantly higher than that in Control group at 21 and 28 days,the difference was statistically significant(P<0.05).The expression of Sesn2 mRNA in the RS group was higher than that in the Control group at 7 and 14 days,but it was not statistically significant.There were no statistical differences in the expression of Bnip3,UCP2,P53,NF-κB,DRP1,E21F,Endog and Pink1 mRNA in the RS group at 7 days,14 days,21 days,and 28 days.(8)Correlation analysis:Correlation analysis of hippocampal plasticity related family protein Plpprs and mitochondrial function-related protein mRNA expression,the expression of plasticity-related family Plpprl was significantly negatively correlated with P53,SENSN2 and PARKIN mRNA expression,and was statistically significant(P<0.05)and there was a significant positive correlation with E21F mRNA expression,and it was statistically significant(P<0.05).There was a significant negative correlation between the plasticity-related family protein Plppr2 and UCP2 mRNA expression,and it was statistically significant(P<0.05).The plasticity-related family protein Plppr3 was significantly negatively correlated with E21F mRNA expression,and was statistically significant(P<0.05).It was significantly positively correlated with Bnip3L,mTOR,HD AC 1,BAD and FUNDC1 mRNA expression,and was statistically significant(P<0.05).There was a significant positive correlation between the plasticity-related family protein Plppr4 and Bnip3L mRNA expression,and it was statistically significant(P<0.05).The plasticity-related family protein Plppr5 was significantly negatively correlated with PINK1 mRNA expression,and was statistically significant(P<0.05).It was positively correlated with Bnip3L and HIF1α mRNA expression,and was statistically significant(P<0.05).(9)Western Blot:①.Plasticity related protein Plpprs:In hippocampus,the expressions of hippocampal plasticity-related proteins Plpprl,Plppr4 and Plppr5 in RS group were significantly higher than those in Control group at 7 days,14 days,21 days and 28 days,and the difference was statistically significant(P<0.05).Cortex,Plpprl:The expression of Plpprl protein in the cerebral cortex of the RS group was significantly lower than that of the Control group at 14 days(P<0.05).The expression of Plpprl protein in the cerebral cortex of the RS group at 21 and 28 days was higher than that of the Control group,and the difference was statistically significant(P<0.05).There was no statistical difference in the expression of Plpprl protein in the cerebral cortex between the RS group and the control group at 7 days.Plppr4:The expression of Plppr4 protein in the cerebral cortex was significantly lower in the RS group than in the control group at 7 and 14 days,the difference was statistically significant(P<0.05);the RS group was associated with cerebral cortical plasticity at 28 days.The expression of Plppr4 in the cerebral cortex of the RS group was significantly higher than that of the Control group at 28 days,and the difference was statistically significant(P<0.05).P1ppr5:The expression of Plppr5 protein in the cerebral cortex of the RS group was significantly higher than that of the Control group at 7 days,14 days,21 days and 28 days,and the difference was statistically significant(P<0.05).②.Mitochondrial function-related proteins:HIF-1 a:The expression of HIF-1 a in hippocampus of RS group was significantly higher than that of Control group at 7 days,14 days,21 days and 28 days,the difference was statistically significant(P<0.05);The expression of HIF-1 a in the cerebral cortex of the RS group was significantly higher than that of the Control group at 7 days,14 days and 28 days,and the difference was statistically significant(P<0.05).The expression of HIF-1αprotein in the cerebral cortex in the RS group was higher than that in the Control group at 21 days,but it was not statistically significant.PGC-1α:The expression of PGC-1α protein in hippocampus of RS group was significantly lower than that of Control group at 7 days,21 days and 28 days,the difference was statistically significant(P<0.05).The expression of PGC-1α protein in hippocampus of RS group was lower than that of Control group at 14 days,but it was not statistically significant.The expression of PGC-1α protein in the cerebral cortex of the RS group was significantly lower than that of the Control group at 7 days,14 days,21 days and 28 days,and the difference was statistically significant(P<0.05).Sirt3:The expression of Sirt3 protein in hippocampus of RS group was lower than that of Control group at 14 and 28 days,the difference was statistically significant(P<0.05).There was no statistical difference in the expression of Sirt3 protein in hippocampus between RS group and control group at 7 and 21 days.The expression of Sirt3 protein in the cerebral cortex of RS group was significantly lower than that of Control group at 7 days,14 days and 28 days,the difference was statistically significant(P<0.05).The expression of Sirt3 in the cerebral cortex of the RS group was lower than that of the Control group at 21 days,but it was not statistically significant.Prohibitin:The expression of Prohibitin in the hippocampus of the RS group was significantly lower than that of the Control group at 14 days,21 days and 28 days,and the difference was statistically significant(P<0.05).There was no statistical difference in the expression of Prohibitin in the hippocampus between the RS group and the Control group at the 7-day time point.The expression of Prohibitin in the cerebral cortex of the RS group was significantly lower than that of the Control group at 7 days,14 days,21 days and 28 days,and the difference was statistically significant(P<0.05).Conclusion:Long-term development,neurobehavior,cognition and hippocampal morphological damage caused by penicillin repeated seizures may be related to the expression and interaction of hippocampal and cerebral cortex plasticity-related protein molecules,mitochondrial function-related protein molecules and zinc ion transporter molecules.Part Ⅱ Ketone diet intervention for long-term plasticity-related protein molecules and mitochondrial function-related protein molecule expressionObject:To investigate the role of hippocampus and cerebral cortex plasticity-associated protein molecules,mitochondrial function-related protein molecules and zinc ion transporter molecules in neuroprotection of ketogenic diet during developmental convulsive brain injury.Method:100 SD rats(postnatal days 21,P21)were randomly divided into 2 groups:Control group(n=40)and modeling group(n=60).Rats in the modeling group developed a penicillin-induced developmental epilepsy model:penicillin was injected intraperitoneally in each rat at a dose of 5 million units/kg/time,once every other day,and a total of 6 injections were performed(P21-31).According to the inclusion criteria,40 moded group(n=40)were obtained.The control rats were intraperitoneally injected with the same dose of physiological saline at the same time as the model group treatment.After the modeling was completed,the control group was further divided into Control group(n=20)and KD group(n=20),and the modeling components were RS group(n=20)and RS+KD group(n=20).The day after the modeling was completed(P32),the KD group and the RS+KD group were given a ketogenic diet for 28 days(P32-P59),and the control group and the RS group continued to consume the normal rat pellets.All rats did not restrict drinking water and the body weight of the rats was detected every other day.By monitoring the development of neuroreflex,emotion,learning and memory index changes;Neo-Timm’s staining to observe hippocampal mossy fiber germination and amygdala Timm staining;Nissl staining to observe the number and morphology of hippocampal and amygdala neurons;Golgi staining to observe neuron axons Length,grading and number of axons;Real-Time RT-PCR detection to observe mRNA expression of hippocampal plasticity related protein family Plpprs,zinc ion channel protein ZnTs and mitochondria-associated protein;Western-Blot was used to detect the expression of plasticity related protein family Plpprs and mitochondria-associated protein expression in hippocampus and cerebral cortex;EMSA was used to detect the transcriptional activity of hypoxia-inducible factor HIF-1α in hippocampus and cerebral cortex.Results:(1)Body weight:The overall difference in the weight of the four groups during the experiment was statistically significant.There was no significant difference in weight between the four groups of P21-P23.The weight of the four groups began to differ from P25.The weight of RS group and RS+KD group was significantly lower than that of Control group,the difference was statistically significant(P<0.05);The weight of the KD group was significantly lower than that of the Control group from P35,and the difference was statistically significant.This difference continued to P59.The weight of RS+KD group was significantly lower than that of RS group from P39,the difference was statistically significant(P<0.05),and the difference continued to P59.In the comparison of the four groups of daily weight gain rates,the daily average weight gain rate of the KD group,the RS group and the RS+KD group was significantly lower than that of the Control group,the difference was statistically significant(P<0.05),KD group and RS+KD group was significantly lower than the RS group,and the difference was statistically significant(P<0.05).(2)Neurodevelopment:In the forelimb suspension test,the fore limb suspension reflex time in the RS group was significantly lower than that in the Control group,the difference was statistically significant(P<0.05);the RS+KD group forelimb suspension reflex time was significantly higher than RS The difference was statistically significant(P<0.05);there was no significant difference from the Control group,which tends to be normal.In the negative geotaxis test,the negative geotaxis reflex time of the RS group was significantly higher than that of the Control group,and the difference was statistically significant(P<0.05).The negative geotaxis reflex time of the RS+KD group was significantly lower than that of the RS group.It was statistically significant(P<0.05).In the cliff avoidance test,the cliffa voidance reflex time of RS group was significantly higher than that of Control group,the difference was statistically significant(P<0.05),and the avoidance reflex time of RS+KD group was significantly lower than that of RS group.The difference was statistically significant(P<0.05).In the planar righting test,the plane righting reflection time of the RS group was higher than that of the Control group,but the difference was not statistically significant;(3)Learning and memory:In the new object recognition experiment,the cognitive index of RS group was significantly lower than that of Control group,the difference was statistically significant(P<0.05);the cognitive index of RS+KD group was significantly higher than that of RS group,the difference was statistical Academic significance(P<0.05);no significant difference with Control group,tending to normal;(4)Blood glucose and blood ketone levels:The blood glucose levels in the KD group and the RS+KD group were significantly lower than those in the Control group and the RS group(P<0.05).There was no significant difference in blood glucose levels between the RS group and the Control group.The blood ketone levels in the KD group and the RS+KD group were significantly higher than those in the Control group and the RS group(P<0.05).There was no significant difference in blood ketone levels between the RS group and the Control group.(5)Neo-Timm’s staining:The analysis of hippocampus in Neo-Timm’s staining showed that the number of Timm particles stained in CA3 and DG areas of RS group was significantly higher than that of Control group and KD group,the difference was statistically significant(P<0.05);The number of stained Timm particles in the CA3 and DG areas of the RS+KD group was significantly lower than that of the RS group,and the difference was statistically significant(P<0.05).There was no significant difference between the Control group and the KD group.By analyzing the amygdala area in Neo-Timm’s staining,the optical density of Timm staining in RS group and RS+KD group was significantly higher than that in Control group and KD group,the difference was statistically significant(P<0.05);The optical density in RS+KD group was significantly lower than that of the RS group,and the difference was statistically significant(P<0.05).(6)Nissl staining:In the Nissl staining of the hippocampus,the number of neurons in the hippocampal CA1 area and CA3 area was significantly lower than that in the Control group and the KD group,the difference was statistically significant(P<0.05),and the neuron structure was disordered.The cytoplasmic Nissl staining was uneven;the number of neurons in the hippocampal CA1 area and CA3 area of RS+KD group was significantly higher than that of RS group,the difference was statistically significant(P<0.05).The number of neurons in the amygdala of the RS group was significantly lower than that of the Control group and the KD group in the Nissl staining of the amygdala.The difference was statistically significant(P<0.05),and the neuronal structure was disordered,and the cytoplasmic Nissl staining was not uniformity;the number of neurons in the amygdala group was significantly higher in the RS+KD group than in the RS group(P<0.05).(7)Golgi staining:This paper used Golgi staining and Sholl analysis to analyze the complexity of cerebral cortical neurons.It was found that the KD group,RS group and RS+KD group had significantly lower neuron protuberances than the Control group,and the difference was statistically significant(P<0.05),the branch level of neurons in the RS+KD group was significantly higher than that in the RS group,and the difference was statistically significant(P<0.05).In the analysis of the length of cerebral cortical neurons,we found that the length of neurons in the RS group was significantly lower than that in the Control group,the difference was statistically significant(P<0.05),and the length of neurons in the RS+KD group was higher than that in the RS group.But the difference was not statistically significant.In the analysis of the number of spins on the dendrites of cerebral cortical neurons,we found that the number of spinous in the RS group was significantly lower than that in the Control group,the diffrerence was statistically significant(P<0.05),and the ketogenic diet could significantly improve this.The number of spins in the RS+KD group was significantly higher than that in the RS group,and the difference was statistically significant(P<0.05).(8)Real-Time RT-PCR:①.Plasticity-related protein Plpprs gene:RS group Plppr3,Plppr4,Plppr5 mRNA expression was significantly higher than that of Control group,the difference was statistically significant(P<0.05);The expression of Plppr3,Plppr4 and Plppr5 mRNA in RS+KD group was significantly lower than that in RS group,the difference was statistically significant(P<0.05).The expression of Plpprl and Plppr2 mRNA in RS group was higher than that in Control group,but it was not statistically significant.There was no statistical difference in the expression of the plasticity-related protein Plpprl mRNA in the RS+KD group compared with the Control group;②.ZnTs:The expression of ZnT1 mRNA in RS group was significantly higher than that in Control group(P<0.05).The expression of ZnTl mRNA in RS+KD group was significantly lower than that in RS group(P<0.05).There was no statistical difference between the RS+KD group and the control group,which was normal.③.Mitochondrial function-related genes:The expressions of Bnip3L,HIF-1α,HDAC1,mTOR and Parkin mRNA in RS group were significantly higher than those in Control group,the difference was statistically significant(P<0.05).The expressions of Bnip3L,HIF-la,HDAC1,mTOR and Parkin mRNA in RS+KD group were significantly lower than those in RS group,the difference was statistically significant(P<0.05).There was no statistical difference between the RS+KD group Bnip3L,HIF-la,HDAC1,mTOR and Parkin mRNA expression compared with the Control group.There was no statistical difference between the RS group Bnip3,UCP2,P53,NF-κB,DRP1,E21F,Endog and Pinkl mRNA expression compared with the Control group.(9)Western Blot:①.Plpprl:In hippocampus,the expression of Plpprl in hippocampus of RS group was significantly higher than that of Control group(P<0.05).The expression of Plpprl protein in hippocampus of RS+KD group was significantly lower than that of RS group.The difference was statistically significant(P<0.05);there was no statistically significant difference between the expression of Plpprl protein in hippocampus of RS+KD group and Control group.In the cerebral cortex,the expression of Plpprl protein in the cerebral cortex of RS group was significantly higher than that in Control group(P<0.05).The expression of Plpprl protein in cerebral cortex of RS+KD group was significantly lower than that in RS group.The difference was statistically significant(P<0.05);there was no statistically significant difference between the expression of Plpprl protein in the cerebral cortex of RS+KD group and Control group.②.Plppr5:In hippocampus,the expression of Plppr5 protein in hippocampus of KD group,RS group and RS+KD group was significantly higher than that of Control group(P<0.05).The expression of Plppr5 protein in hippocampus of RS+KD group was better than RS.The group was significantly reduced,and the difference was statistically significant(P<0.05).In the cerebral cortex,the expression of Plppr5 protein in the RS group and RS+KD group was significantly higher than that in the Control group(P<0.05).The expression of Plppr5 protein in the RS+KD group was significantly lower than that in the RS group.Statistically significant(P<0.05).③.HIF-la:In hippocampus,the expression of HIF-la in hippocampus of KD group,RS group and RS+KD group was significantly higher than that of Control group,the difference was statistically significant(P<0.05).The expression of HIF-1α in hippocampus of RS+KD group was significantly lower than that of RS group,and the difference was statistically significant(P<0.05).In the cerebral cortex,the expression of HIF-la protein in RS group was significantly higher than that in Control group(P<0.05).The expression of HIF-1αprotein in cerebral cortex of RS+KD group was significantly lower than that in RS group(P<0.05),there was no statistical difference in the expression of HIF-la protein in the cerebral cortex between the RS+KD group and the control group.④.PGC-1α:In hippocampus,the expression of PGC-1α protein in RS group was significantly lower than that in Control group(P<0.05).The expression of PGC-1α protein in hippocampus of KD group and RS+KD group was significantly higher than that of Control group and RS group(P<0.05).In the cerebral cortex tissues,the expression of PGC-1αprotein in KD group,RS group and RS+KD group was significantly lower than that in Control group,the difference was statistically significant(P<0.05).The expression of PGC-1α protein in the cerebral cortex of RS+KD group was significantly higher than that in RS group(P<0.05).⑤.Sirt3:In hippocampus,the expression of Sirt3 protein in RS group was significantly lower than that in Control group and KD group(P<0.05).The expression of Sirt3 protein in hippocampus of RS+KD group was significantly higher than that in RS group(P<0.05);there was no statistical difference between the expression of Sirt3 protein in hippocampus of RS+KD group compared with Control group.In the cerebral cortex tissue,the expression of Sirt3 protein in the cerebral cortex of RS group was significantly lower than that of Control group and KD group(P<0.05).The expression of Sirt3 protein in the cerebral cortex of RS+KD group was significantly higher than that of RS group(P<0.05).⑥.p-AMPK:In hippocampus,the phosphorylation level of AMPK protein in hippocampus of RS group was significantly higher than that of Control group,the difference was statistically significant(P<0.05).The phosphorylation level of AMPK protein in hippocampus of RS+KD group was significantly lower than that of RS group(P<0.05).There was no statistically significant difference between the phosphorylation level of hippocampal AMPK protein in RS+KD group and Control group.In the cerebral cortex tissues,the phosphorylation level of AMPK protein in the cerebral cortex of KD group and RS group was significantly higher than that of Control group,the difference was statistically significant(P<0.05).The phosphorylation level of AMPK protein in cerebral cortex of RS+KD group was significantly lower than that of RS group(P<0.05).The phosphorylation level of AMPK protein in cerebral cortex of RS+KD group was not statistically significantly different from that of Control group.⑦.Bnip3L:In hippocampus,the expression of Bnip3L protein in hippocampal of RS group was significantly higher than that of Control group(P<0.05).The expression of Bnip3L protein in hippocampus of RS+KD group was significantly lower than that of RS group(P<0.05).There was no statistically significant difference between the expression of Bnip3L protein in hippocampus of RS+KD group and Control group.In the cerebral cortex,the expression of Bnip3L protein in the cerebral cortex of RS group was significantly higher than that in Control group(P<0.05).The expression of Bnip3L protein in cerebral cortex of RS+KD group was significantly lower than that in RS group(P<0.05).There was no statistically significant difference in the expression of Bnip3L protein in the cerebral cortex between the RS+KD group and the Control group.⑧.Prohibitin:In hippocampus,the expression of Prohibitin in hippocampus of KD group and RS group was significantly lower than that of Control group(P<0.05).The expression of Prohibitin in hippocampus of RS+KD group was significantly higher than that of RS group(P<0.05).There was no statistically significant difference between the expression of Prohibitin in the hippocampus of RS+KD group and Control group.In the cerebral cortex tissues,the expression of Prohibitin in the cerebral cortex of the KD group and the RS group was significantly lower than that of the Control group(P<0.05).The expression of Prohibitin in the cerebral cortex of the RS+KD group was significantly higher than that of the RS group.The significance of learning(P<0.05);there was no statistically significant difference between the expression of Prohibitin in the cerebral cortex of RS+K1D group and Control group.(10)EMSA:The transcriptional activities of hippocampus and cortex HIF-1α in KD group,RS group and RS+KD group were significantly higher than those in Control group,the difference was statistically significant(P<0.05).Conclusion:The neuroprotective effect of ketogenic diet on developmental convulsive brain injury may be achieved by regulating hippocampal and cerebral cortex plasticity-associated protein molecules,mitochondrial function-related protein molecules and zinc ion transporter molecules.Part III Protective effect of ketone bodies on glutamate toxicity in vitro cell model and mechanism of Plppr5 neuroprotectionObject:To investigate the protective effect of ketone bodies on glutamate toxicity in vitro and the mechanism of Plppr5 neuroprotection.Method:Quarter 1:The mouse hippocampal neuron cell line HT22 was divided into four groups according to the treatment:Control group,P-HB group,Glutamate group and Glutamate+β-HB group.The definition and treatment of each group were as follows:Control group:blank control group,cells were not treated;β-HB group:ketone body β-HB(β-hydroxybutyric acid,pH 7.4)treated cells for 24 h(5mmol/L);Glutamate group:glutamate treated cells for 24h(5mmol/L,pH7.4);Glutamate+β-HB group:glutamate(5mmol/L)and β-HB(5mm... |
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