| Objective : Intracranial hemorrhage(ICH)is a common disease in neonatal period infants,which is associated with its anatomical physiology characteristic and variety of perinatal high-risk factors.Germinal matrix-intraventricular hemorrhage(GM-IVH)is the most common type of intracranial hemorrhage.The gestational age is smaller,the incidence of GM-IVH is higher.And it is one of main reasons for affenting the Timely rescue,leading premature to die and even getting a poor prognosis.In recent years,due to the progress of medical technology,especially the improvement of equipments and medical care of the neonatal intensive care unit,the incidence of ICH has fallen to about 2.21%,but the incidence of gestational age < 34 is still as high as 24.1%.Grade ? or ? GM-IVH can cause complications like Periventricular Leukomalacia and Periventricular Hemorrhagic infarction.So far there is no effective treatments.The hippocampus,as an important part of the limbic system,is closely related to the ventricular system.Its Main function is to study and cognize.It is the Potential injury place of GM-IVH.Lesions in the hippocampus is closely related to the variety of nerve mental illness.At present,most of the researchs are still focusing on white matter damage,few people observe whether GM-IVH cause damage to the hippocampus.This purpose of this study wants to explore two questions.Do GM-IVH will affects the expression level of apoptosis-related protein in the hippocampus? Do treatment with deferoxamine will affect the expression level of apoptosis-related protein in the hippocampus?Methods:1 Premature rabbits were born by cesarean section and given incubated,milk powder feeding after born,Then the premature rabbits were randomlydivided into three groups,control group,model group and treatment group,each group had twelve premature rabbits.With reference to the method of intra-peritoneal injection which induces GM-IVH.Model preparation method was as follows:the premature rabbits in GM-IVH group and treatment group were administered 50% glycerol(10ml/kg)at 3 hour of age.After injection of glycerol 2 hours,the treatment group were administered deferoxamine(50mg/ml)for total dose of 100 mg/(kg·d),two times within 24 h.The rest of the two groups were given injection of the same dose of saline.After cerebral ultrasound screening at 24 hour of age,to get rid of the rabitts without GM-IVH in model group and treatment group,the rabbits combine with spontaneous GM-IVH in normal group were also been dropped.2 All the premature rabbits were administered with 2% pentobarbital sodium(40mg/kg),anesthetized and beheaded at the age of 48 hours.Separating the hippocampus on the ice in a very short period of time.One part was immersed in 4% paraformaldehyde solution for the immunohistochemical experiment,the other part was putted in-80? refrigerator for the use of western blot,3 To find the changes between neurons in hippocampus CA1 region under the microscope.To observe the expression of TNF-a and caspase 3protein by immunohistochemical staining.To assess the levels of protein expression of TNF-a and caspase 3 by Western Blot detection.4 Statistical analysis SPSS 17.0 statistics software and Microsoft Excel2003 were used to carry on statistics processing.If the data conforms to normal distribution,which was performed with mean ± standard deviation.If the data are homogeneity of variance,the comparison between each group were performed with LSD.If the data do not meet the homogeneity of variance,use rank sum.P-value<0.05(double side)had statistical significance.Results:1 We performed a cesarean section to deliver the pups prematurely at embryonic day(E)E29(full-term,E32).The head ultrasound screening and specimens showed that the models of GM-IVH was successfully reproducedin model group and treatment group.2 By HE staining,we found the differents between the neurons in each group of hippocampus CA1 region.The neurons in control group were arranged neatly,their structure was clear.The shape of the neurons was vertebral shape.The cell nucleus were big and round,their nucleolus were obvious.Intercellular space of model group was expanded.Cell volume of neurons in model group was smaller.The nucleus of neurons in model group were stained deeper.Some cellular structure were even collapsed.The Nerve cell number in treatment group was between control group and model group.The cellular morphology in treatment group was basic within the normal range.3 The neurons were rich,and there were a few positive cells in hippocampus CA1 region of control group.Compared with control group,there were a large number of positive cells in hippocampus CA1 region of model group,the cytoplasm and nucleus were dyed brown or brown,the difference reached statistical significance.Compared with model group,the number of positive cells were Little,the dyeing was shallow,the difference reached statistical significance.4 The expression of TNF-? protein and caspase-3 protein was little in control group.Compared with the control group,the expression of TNF-?protein and caspase-3 protein was higher in model group after GM-IVH,difference was statistically significant.Compared with the model group,the expression of TNF-? protein and caspase-3 protein was less in treatment group after treatment with DFX,difference was statistically significant.Conclusions:1 After GM-IVH,The number of neurons in hippocampus CA1 region was reduced,the cellular structure was damaged.It reveal that GM-IVH can cause damage to the hippocampus.2 The expression of TNF-? protein and caspase-3 protein was high in model group after GM-IVH,the difference reached statistical significance.It reveal that apoptosis had participated in the process of damage.3 After treatment with DFX,the expression of TNF-? protein and caspase-3 protein was less in treatment group compared with the model group,the difference reached statistical significance.It indicated that DFX has the nerve protective effect. |
PDF Full Text Request