| Objective:Based on TMT relative quantitative proteomics technology,to detect differentially expressed proteins in the hippocampus of rats with and without cognitive impairment in DM,and perform biological information on the differentially expressed proteins To analyze the differential expression of DM cognitive impairment and its relationship with disease mechanism.Methods:106 male Wistar rats were randomly divided into a control group（n=10）and a DM model group（n=96）.In the DM model group,60mg·kg-1of streptozotocin（STZ）was injected intraperitoneally to establish a DM rat model.The glucose oxidase method was used to measure the fasting blood glucose for 72h.The fasting blood glucose value was≥16.7 mmol·L-1.Modeling standards:Dynamically monitor fasting blood glucose in rats with a blood glucose meter at 0d（before modeling）and at 7d,20d,35d,60d,and 80d after modeling;Respectively,at 0w（Before modeling）,the body weight and water consumption of rats were monitored dynamically from 1 to 12 weeks after modeling;Morris water maze was used to detect the learning and memory abilities of rats at 84 days after the modeling,and the learning performance of the rats was individually evaluated to identify DM cognitive and non-cognitive impairment rats.TMT relative quantitative proteomics technology was used to detect differentially expressed proteins in the hippocampus of each group of rats,and GO functional annotation and KEGG pathway annotation were used to analyze the differentially expressed proteins for bioinformatics.Results:Compared with the control group:the fasting blood glucose of the DM model group was significantly increased at 72h,7d,20d,35d,60d,and 80d（p<0.01）,and the body weight began to decrease significantly at 1w（p<0.01）.And continued to 12w,drinking water began to increase significantly at 1w（p<0.01）and continued to 12w.In the Morris water maze experiment,compared with the control group,the swimming speed of the DM model group had no significant difference（p>0.05）,and the escape latency of the 5th to 9th training sessions was significantly increased（p<0.01,p<0.05）,the number of crossing the platform decreased significantly（p<0.01）.Compared with the first training:the learning performance of the rats in the control group was significantly improved at the beginning of the sixth training（p<0.05）,while the learning performance of the rats in the DM model group did not significantly improve during the entire training process（p>0.05）.The cognitive and non-cognitive impairment learning and memory evaluation results of rats in each group showed that the incidence of cognitive impairment in the control group was zero;the incidence of cognitive impairment in the DM model group was 46.9%.This experiment was based on TMT relative quantitative proteomics technology to detect hippocampal tissue proteins in each group of rats.The results showed that 53 differentially expressed proteins were detected in the hippocampal tissue of the DM non-cognitive impairment group and the control group,and the expression was up-regulated.There are 34 differential proteins and 19 differentially down-regulated proteins.A total of 6 differentially expressed proteins were detected in the hippocampus of DM cognitive and non-cognitive damaged rats.Among them,proteins whose expression was up-regulated were:Rab,member of Ras oncogene family like 2,flavin containing monooxygenase1,calcyphosine-like,histone cluster 1 H1 family member a,2,4-dienoyl-Co A reductase 1 and enolase 3.GO function enrichment analysis of differentially expressed proteins in hippocampus of DM cognitive and non-cognitive damaged rats shows that biological processes mainly involve:photoreceptor cell morphogenesis,pyridine nucleotide metabolic process,nicotinamide nucleotide metabolic process,pyridine-containing compound metabolic process,oxidoreduction coenzyme metabolic process,NADPH oxidation,organic acid metabolic process and toxin metabolic process.The enrichment analysis of differentially expressed protein KEGG pathway in hippocampus of DM cognitive and non-cognitive rats showed that the metabolic pathway mainly involves:Drug metabolism-cytochrome P450,Glycolysis/Gluconeogenesis,RNA degradation and HIF-1 signaling pathway.Among them,the drug metabolism-cytochrome P450 metabolic pathway appeared in two pairs of comparison groups.Conclusion:Based on TMT relative quantitative proteomics technology,six differentially expressed proteins were detected and screened in the hippocampus of DM cognitive and non-cognitive impairment rats,among which Rab,member of Ras oncogene family like 2,calcyphosine-like,histone cluster 1 H1 family member a,2,4-dienoyl-Co A reductase 1 and enolase 3 may be protein markers for early diagnosis of DM cognitive impairment.Biological processes involved in differentially expressed proteins suggest that DM cognitive impairment may have abnormal redox reaction processes in hippocampal tissue cells,causing abnormal cell energy metabolism and impairing cognitive function.The metabolic pathways involved in differentially expressed proteins suggest that abnormalities in the glycolysis/gluconeogenesis pathway and HIF-1 signaling pathway may exist in the hippocampus of DM cognitively impaired rats,inhibit the tricarboxylic acid cycle,and cause abnormal energy metabolism in the hippocampus,resulting in The occurrence of DM cognitive impairment. |
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