| Background and PurposeChildren with congenital neurogenic bladder mainly due neural tube malformation underwent surgery after nerve damage caused. Further, since the poor regenerative capacity of the nervous system, in the treatment of a medical face enormous challenges. Since neurogenic bladder traditional treatment less effective, as well as neuronal damage can not be reversed in this pathogenesis. Currently many animal models of spinal cord injury, but there are many drawbacks. Thus, the treatment of neurogenic bladder lack of animal models. Also, because the treatment of neurogenic bladder can not rely solely on clinical research exploring treatment. Therefore, a suitable animal model of neurogenic bladder related more important. Because children with neurogenic bladder caused mainly due to neural tube defects, so we need to find a reasonable animal model of neural tube defects. According to research reported in Accutane, cyclophosphamide, methotrexate, valproate was induced in mice, rats spinal cord injury. However, mice, rats, are not conducive to further treatment of neurogenic bladder study, therefore, to find a beneficial therapy neurogenic bladder in animal models become imperative. In recent years, stem cell transplantation research to the medical problems brought some hope. According to relevant reports spinal cord NSE, AchE, NFP, s-100expression after spinal cord injury with neurogenic bladder is formed with a certain relationship, we can determine the above indicators spinal cord injury.We explore the spinal cord injury with neurogenic bladder animal relationships, seeking the closest child pathogenesis animal models of spinal cord injury, and to find a reasonable animal model of neurogenic bladder production methods. Animal models by detecting spinal cord AchE, s-100, NSE, NFP expression, to understand their relationship with spinal cord injury, spinal cord injury and thus understanding of the drug led to the formation of neurogenic bladder relationship.Object and MethodOur preliminary experiments with SD rats, select New Zealand pregnant rabbits as experimental animals.SD rat animal model:select the newly identified pregnant SD rats were randomly divided into35groups of cyclophosphamide, methotrexate group, retinoic acid group, sodium valproate group and control group. Each experimental group completed in accordance with the preparation of reagents in rats after pregnancy ninth day every day16:00to give the corresponding drugs. Normal control group received normal saline injection.New Zealand pregnant rabbits animal model:Choose OK pregnant New Zealand rabbits were randomly divided into the cyclophosphamide group, sodium valproate group, methotrexate group, retinoic acid group and control group. Each experimental group completed in accordance with the preparation of reagents at16:00every day since the fifth day after pregnancy to give the corresponding drugs. Normal control group received normal saline injection.Immunohistochemical observation of fetal spinal cord:normal childbirth survival after checking fetal rabbit, check the appearance of the embryo to determine whether there is the appearance of visible deformities and record the results. Abortion, abortion pregnant rabbits given timely recording time, collecting abortion fetal rabbit, check whether the appearance of visible deformities. In addition, for pregnant rabbits given anatomical aborted temporarily give frozen, to be removed after the reunification of specimen collection complete spinal cord HE staining and immunohistochemistry. Given continued feeding pups survived the death of pups take spinal fixed, paraffin-embedded sections for further immunohistochemical observation spinal cord acetylcholinesterase (AchE), S-100protein, neuron-specific enolase (NSE) and nerve silk protein (NFP) levels.Data processingSPSS17.0statistical software to complete the data processing. Experimental data to±s, Kolmogorov-Smirnov normality test, the data is normally distributed, multi-group variance (ANOVA) was used to compare the analysis and Bonferroni posthoc analysis, t-test was used to compare the group; data is non-positive state distribution, rank sum test (H test). P<0.05was considered significant meaning.Experimental ResultsSD rat animal model:each experimental group were pups lumbosacral skin defect. The group of cyclophosphamide and methotrexate group were pregnant rats miscarriage, abortion rate were39.13%and16.67%, appearance no abortion pups lumbosacral skin defects. Experimental group, cyclophosphamide and retinoic acid group teratogenic relatively high rates of13.04%and14.81%, respectively. Each experimental group in the cyclophosphamide group, the the methotrexate group of pups mortality high of60.87%and54.17%, respectively. The control group did not appear Rat Abortion, the of farrowing mice were no lumbosacral skin defects. We are poor the experimental pups after all rats anatomy of spinal cord integrity.New Zealand rabbit animal model:cyclophosphamide, methotrexate, retinoids group have pups lumbosacral skin defects. Cyclophosphamide group pups total of21, the mortality rate of76.19%, teratogenic rate of9.52%; the methotrexate group of19total pups, the mortality rate of57.89%, teratogenic rate of10.53%; BingXu sodium group communist pups21, the mortality rate of47.62%, teratogenic rate of0. Communist pups retinoic acid group24, the mortality rate of45.83%, teratogenic rate of12.5%. Fetal rat saline control group all survived no significant lumbosacral skin defects.Immunohistochemistry showed that:the average gray integrated optical density value of fetal rabbit spinal cord AchE (acetylcholinesterase), s-100protein, NSE (nerve-specific enolase), NFP (anti-neurofilament protein) positive (G) with saline as compared to the control group was statistically significant (P<0.05). We found acetylcholinesterase, neuron-specific enolase G compared with the control group decreased (p<0.5) was statistically significant; s-100protein and neurofilament protein is elevated (p<0.5), statistics significance. Cyclophosphamide group, valproate group, methotrexate group and retinoic acid group group variance (ANOVA) analysis was no significant difference (P>0.05).Experimental Conclusion1. Cyclophosphamide, sodium valproate, methotrexate, and all-trans retinoic acid in the rat spinal cord injury in animal models of spinal cord inhibition. Including cyclophosphamide, all-trans retinoic acid inhibition of rat spinal cord and methotrexate compared with sodium valproate is relatively obvious, but the abortion rate is higher, lower teratogenicity.2. Cyclophosphamide, sodium valproate, methotrexate, and all-trans retinoic acid on rabbits have a certain degree of spinal cord injury, but showed no appearance of deformities. New Zealand pregnant rabbit’s abortion rate is higher, relatively high mortality of pups, teratogenic rate is low. Among four groups cyclophosphamide, all-trans retinoic acid on rabbits and valproate compared with methotrexate has better spinal cord injury.3. The experimental group Zealand rabbit spinal cord acetylcholinesterase, s-100protein, neuron-specific enolase, neurofilament protein compared with the normal group had some changes. AchE and NSE in animal models have a certain degree of reduction, and s-100protein and neurofilament protein was increased to some extent, there is a certain relationship with spinal cord injury. |
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