| ObjectivesNeural tube defects(NTD) are a common congenital malformation in humans, which have affected on the human quality and are the most severe desease of birth defects,which comprises a series of defects such as anencephalia,encephalocoele, myeloschisis and so on because of no and delay closure of the neuropore in the progress of developing embryo.Apoptosis is as much a part of embryonal development as is cell proliferation and differentiation.This cell suicide is controlled by cell genes involved in induction or prevention of programmed cell death.During embryogenesis apoptosis implicates cell elimination,which is necessary for fashioning of the body and moulding of the tissues. An exaggerated or a defective apoptosis can perturb the normal development and morphogenesis of individual organisms and organs and cause various developmental abnormalities.Caspase-3 is an active cell death protease involved in the execution phase of apoptosis.Its null mutant mice exhibits a remarkablely NTD that has been attributed in part to a failure of apoptosis of neurons.Apoptosis can rise in Neural tube defects rat,which is raised by some aticles.At present,there are many accesses to explain the mechanism about apoptosis,among which the caspase protease family plays critical rlies in the apoptosis through death receptors.This access can sequentially activate caspase-4,caspase-7,caspase-8, caspase-3,caspase-6 and eventually induces cell apoptosis.It is found that caspase-3 knocked-out mouse tend to dead early,which tells us that caspase-3 plays the important role in neuronal apoptosis.Mitogen-activated protein kinase(mitogen-activated protein kinase,MAPK) are eukaryotic signal transduction network in one of the important ways.Has been found that there are a number of parallel MAPK signal transduction pathway,different extracellular stimulation can activate different MAPK signaling pathway,many growth factors,cytokines,G protein-coupled receptor,stress signals and mitogen-all can activate MAPK signal transduction pathway.MAPK signal transduction pathway in cell proliferation,differentiation and apoptosis plays a key role.Five kinds of mammals at different MAPK signal transduction pathway,c-Jun N-terminal kinase(c-Jun NH2-terminal kinase,JNK) and P38MAPK signal transduction pathway in inflammation and apoptosis,such as stress response play an important role.On this basis,this thesis aims to explore the neural tube defects in fetal rat spinal cord tissue cell apoptosis,while analysis of apoptosis gene Caspase-3 and JNK relations.In this study,12,15,17 and 20 of embryonic malformations in rats as the research object,the establishment of neural tube defects in rat animal model,using HE staining,immunohistochemical staining method,Western-blot method to observe embryonic 12,15,17 and 20 abnormalities in rats nerve tissue cells caspase-3 expression and distribution,and further to the caspase-3 expression and MAPK phosphorylation JNK family relations were discussed.Method1.Rat model of neural tube defects and animal production sub1)retinoic acid dissolved in olive oil(40mg/ml),to a dose of 135mg/kg administered by stomach tube into the 10-day pregnant rats produced Wister rat model of neural tube defects,equivalent to the olive oil control group.2) Experimental sub-groups:the control group and the Spina Bifida Unit 12 days in accordance with the pregnancy,15 days,17 days and 20 days divided into four subgroups.2.Spina bifida malformation group fetal rat spinal cord tissue analysis of apoptosis and apoptosis signaling pathway in the caspase-3 and MAPK family,the relationship between JNK1)the application of immunohistochemical methods caspase-3 in the control group, fetal malformation group in the rat spinal cord tissue distribution and expression.2)the application of Western blot used to detect abnormal group and the control group fetal rat spinal cord tissue MAPK phosphorylated JNK family situation. Result1.First,rat model of neural tube defectsAt 10 days pregnant rats given retinoic acid(135mg/kg body weight), progesterone 12 days,15 days,17 days,20 days fetal rat out section can be found encephalocele,spina bifida,no tail,anal atresia and other deformity.2.12-day rat embryo,15 days,17 days,20 days in control group and the Spina Bifida Group of spinal cord nerve tissue malformation caspase-3 expression analysisImmunofluorescent staining method used can be found in embryos of 15 days, 17 days and 20 days of dominant malformation spina bifida rat spinal cord tissue caspase-3 positive cells than the control group,fluorescence intensity of the control group.Embryos within 15 days,17 days,20 days of dominant malformation spina bifida rat spinal cord tissue caspase-3 positive cells were more than 12 days of embryonic malformation spina bifida dominant group3.12-day rat embryo,15 days,17 days,20 days in control group and the group of spina bifida malformation of the spinal cord nerve tissue JNK phosphorylation of MAPK family situationWestern-blot method with a different group Detected fetal rat spinal cord nerve tissue in the MAPK family,JNK phosphorylation,results showed that the 12-day embryo,15 days,17 days and 20 days malformation spina bifida rat spinal cord nerve tissue higher than the level of JNK phosphorylation.Spina bifida malformation rat spinal cord 15 days embryo nerve tissue levels of JNK phosphorylation peaked.Conclusion1.12,15,17,20 days embryo dominant spina bifida fetal rat spinal cord tissue caspase-3 positive cells in the number of 15 days beginning from the embryo increased, suggesting that the peak phase of cell apoptosis.2.overt spina bifida fetal rat spinal cord nerve tissue cells,JNK phosphorylation of MAPK family embryos at 15 days to reach the peak,while overt spina bifida fetal rat spinal cord tissue caspase-3 positive cells in the number of 15 days beginning from the embryo increased,suggesting that apoptosis caspase-3 protein expression increased with the MAPK family,JNK signaling pathway related to apoptosis of abnormal cells that participate in the retinoic acid-induced spina bifida fetal rat dominant formation process,and this anomaly may be associated with apoptosis MAPK family,JNK apoptotic signal transduction pathway related.
|
PDF Full Text Request