| Objective:To investigate the possible mechanisms of the long-term impact of nervous system in young rats fowllowing intrauterine environment changes, vlidation the differences level of transcription and protein expression on study-related signal pathway, and provide theoretical and experimental basis of the effect of fetal origins of adult disease on the neurodevelopment.Method:On the basis of successfully established malnutrition and preeclampsia animal models,8-week-old offspring of the brain tissue in either two groups of maternal low-protein malnutrition and preeclampsia were both sampled as well as the conctrol group. The location, protein expression and mRNA of NMDA receptors in NMDAR1, NMDAR2B, dopamine receptors in DR1, DR2 and glucocorticoid receptors was determined by immunohistochemisty, Western Blot and Real-time PCR.Results:(1) The results of Immunohistochemical showed that, compared whith the control group, there was decreased level expression as well as the number and distribution of NMDA receptors in NMDAR1, NMDAR2B, dopamine receptors in DR1, DR2 in the low-protein malnutrition group, while there was no difference of glucocorticoid receptors in both groups; compared whith the control group, there was decreased level expression as well as the number and distribution of NMDA receptors in NMDAR1, dopamine receptors in DR1, but increased level expression of glucocorticoid receptors in the preeclampsia group, and there was slightly increased expression of dopamine receptors in DR2 with partial negative expression, while there was no difference of NMDA receptors in NMDAR2B in both groups. (2) Western Blot results suggested the protein expression of NMDA receptors in NMDAR1, NMDAR2B of the low-protein malnutrition group was significantly lower than that of the control group (p<0.05), while there was no significant difference of dopamine receptors in DR1, DR2 and glucocorticoid receptors protein expression levels in both groups. Western Blot results suggested the protein expression of glucocorticoid receptors of preeclampsia group was significantly higher than that of the control group (p<0.05), while there was no significant difference of NMDA receptors in NMDAR1, NMDAR2B, dopamine receptors in DR1, DR2 protein expression levels in both groups. (3) Real-time PCR results suggested the expression of NMDA receptors in NMDAR2B mRNA of the low-protein malnutrition group was significantly lower than that of the control group (p<0.05), while there was no significant difference of NMDA receptors in NMDAR1, dopamine receptors in DR1, DR2 and glucocorticoid receptors mRNA expression levels in both groups. Real-time PCR results also suggested the expression of glucocorticoid receptors mRNA of preeclampsia group was significantly higher than that of the control group (p<0.05), while there were no significant difference of NMDA receptors in NMDAR1, NMDAR2B, dopamine receptors in DR1, DR2 mRNA expression levels in both groups.Conclusion:Intrauterine environment changes lead to the reduction in NMDA receptors and dopamine receptors, while the increased level of glucocorticoid receptors. It may be the mechanism of nervous system disorders in their offspring during adolescence period.Objective:To investigate the mechanism of the impact of fetal origins of adult disease in neurodevelopment and profile the methylation alterations of CpG islands in brain gene of 8-week-old offspring rats fowllowing intrauterine enviornment changes in epigenetic level study, in order to understand and screen the impact of methylation modification.Study design:On the basis of successfully established maternal low-protein malnutrition animal models,8-week-old offspring of the brain tissue in either two groups of maternal low-protein malnutrition and normal were both sampled. Using rat Roche-NimbleGen CpG promoter chips, screening the difference in CpG island methylation of genes, the different genes related with maternal low-protein malnutrition were analyzed by Gene Ontology (GO) and KEGG pathway. Results:(1) The differences in genetic screening are apparently. A total of 18541 genes in the CpG island methylation screened 1181 different genes, with 542 (45.85%) genes hypermethylated and with 639 (54.15%) genes demethylated, while with 1018 (50.70%) genes hypermethylated and with 990 (49.30%) genes demethylated in promoter area. (2) The GO analysis showed:In CpG island there was a category related to memory (p=0.0029), three categories related to neurodevelopment, which were forebrain development (p=5.58E-06), midbrain development (p=0.0032) and neuron maturation (p=0.0342), as well as a category related to neuropeptide signaling pathway (p=0.0047) occurring in the demethylated genes of the experimental group. Both hypermethylation (12 categories,22.64%) and demethylation (19 categories, 13.77%) in the experimental group involved in development and morphogenesis, which sharing a great range of methylated genes. And in promoter area, there were two categories related to neurodevelopment, which were central nerve system development (p=0.0252) and brain development (p=0.0426), and a category related to neuropeptide signaling pathway (p=0.0042) as well as a category related to regulation of neurogenesis (p=0.0336) occurring in the demethylated genes of the experimental group. (3) The result of KEGG searching showed:There were 5 methylation different pathways in CpG island, including neuroactive ligand-receptor interaction (p=0.0028), while 4 methylation different pathways in promoter area, in which neuroactive ligand-receptor interaction, calcium signalling pathway and ECM-recptor interaction highly correlated (p<0.05).Conclusion:The epigenetic statue has changed during pregnancy stage of embryonic development caused by intrauterine environment changes, with a range of key genes related to the changes in CpG island methylation. The differences of methylation levels may lead to the mechanism of long-term nervous system caused by intrauterine envionment changes.
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