| Objectives: To all trans retinoic acid(all- trans retinoic acid, ATRA) induction of neural tube defects of rat model was constructed as the research object, explore the neural tube malformation, neural tube defects, NTDs in the process of the development of H3K27me3(H3 histone three methylation status of lysine27)and folate metabolism related genes Adcy2 and expression changes, understand the effect of folic acid intervention in this process, in the process of the occurrence of neural tube defects prevention effect of folic acid interfere, for the prevention of neural tube defects(neural tube defects, NTDs) provide reliable theoretical basis for early.Methods: 1.The establishment of animal model: The 24 healthy and maturation of female SD rats were divided into three groups at random; 1.1 morphology observation: in E15 d cutting palace extraction, observe the morphological characteristics of fetal rats, record the number of each type of fetal rats, statistics and analysis of incidence of malformation fetal rats. 1.2 electron microscope: from each neural tube, after electron embedding under transmission electron microscope and photographed. 1.3 pathology observation: take each embryos HE staining, and under a microscope the pathological changes.2. H3K27me3 expression of detection: The amount of H3K27me3 expression in fetal rats with immunohistochemical technique method and Western blot technique to detect expression of three groups of H3K27me3 in embryo, and carries on the qualitative and quantitative analysis.3. Determination of Acdy2 expression: The QRT- PCR technology for each Adcy2 gene expression in fetal rats quantity measurement analysis.Results: 1. Produce animal models and observations: 1.1 Statistic analysis showed malformation fetal rats detected in normal group, the experimental group the incidence of neural tube defects is 84%, intervention group the incidence of abnormal folic acid is 41.1%; Experimental deformity rate and higher than those in normal groups, statistically significant difference(P < 0.01), deformity rate increased folic acid intervention group than the normal group, but compared with the experimental group the deformity rate dropped significantly, statistically significant difference(P < 0.05).1.2 Electron Microscope Results: The normal group of neural tube group, its nucleus is complete, a round or ovoid. The mitochondria is clearly visible; Experimental neural tube that is incomplete, the deformation is serious, mitochondria shrinkage, even dissolve; Folic acid intervention group neural tube nucleus deformation, shrinkage phenomenon, although there are also compared with the experimental group, but the conditions have corresponding improvement, intercellular space returns to normal, a small number of mitochondria shrinkage, dissolved.1.3 HE staining results: Normal group of normal fetal rats can see its neural tube development, form a normal spinal cord. Dominant spina bifida of fetal rats in the experimental group, the spinal cord to change complex, spinal dorsal skin interrupt discontinuous, spinal cord damage disorder structure, spinal cord with spina bifida bulging outward, accompanied by inflammatory cells invasion. Folic acid spina bifida in intervention group is less, koala deformity, folic acid intervention group compared with the experimental group development have improved, but still poorer than the normal group development.2. Each group H3K27me3 expression quantity of detection: Immunohistochemical results showed that in the normal way of H3K27me3 expression quantity is low, it is strong positive expression in experimental group, folic acid, amount of its expression in intervention group was slightly lower than that of the experimental group, but did not show on a business trip of the opposite sex, and compared with normal group, the difference is obvious, with statistical significance(P < 0.05). Western blot test results showed that the experimental group in the expression of H3K27me3 folic acid is higher than the normal group and intervention group, the difference was statistically significant(P < 0.05), folic acid quantity between the two groups, intervention group expression differences apparent, with statistical significance(P < 0.05). Immunohistochemical results: low amount of H3K27me3 expression in normal way, it is strong positive expression in experimental group, folic acid, its expression in intervention group was slightly lower than that of the experimental group, but did not show on a business trip of the opposite sex, and compared with normal group, the difference is obvious, with statistical significance(P < 0.05)3. Each group Adcy2 gene QRT- PCR detection results showed that in experimental group increased, compared with normal group and intervention group folic acid, statistically significant difference(P < 0.05); In the amount of folic acid in the interention group expression between the two groups, statistically significant difference(P < 0.05).Conclusion: 1. Folic acid intervention can effectively reduce the occurrence of neural tube defects rate;2. In NTDs rats, the H3K27me3 expression level increases, the expression of folate metabolism related genes Adcy2 rise too, folic acid intervention can effectively reduce the expression of the two. |
PDF Full Text Request