The Role Of NMDA Receptor Activation In The Dystrophy Of The Lung Of Fetal Rats Induced By Intrauterine Hypoxia

ChapterⅠThe effect on fetal lung development in intrauterine hypoxiaObjective The establishment of intrauterine hypoxia model, observation of intrauterine hypoxia on the impact of development in fetal rat,focusing on fetal rat lungMethods 1.Preparation of animal model and experimental To get the model of intrauterine hypoxia rats by decreasing the fraction of inspired oxygen of pregnant rats to 10.5±1.0%,hypoxia lasted eight hours a day.Experimental grouping as follows:air-control group, hypoxic two- day group,hypoxic six-day group and hypoxic 10-day group.Get the fetal rats by opening the uterus on E21.2.General condition results To record pregnant rat weight,fetal rat weight, number of fetal rats,survival rate,fetal heart(right ventricle/left ventricle+septum)(RV/LV + S) and so on.3.Histopathological examination of placental To reserve placental of air-control group and all hypoxia group,together with two cases of umbilical cord and blood vessels,the selection criteria is the largest and the smallest weight group which carried out the relevant checks pathology.4.Lung tissue results To wet the fetal rat lung weight,check the left part of fetal lung for HE staining,radial alveolar count(RAC),the pulmonary artery diameter, wall thickness,wall thickness/pulmonary arterial diameter(WT%),the total area of vascular wall area,wall area/total area(WA%).5. Statistical analysis Test data indicated that in mean±standard,deviation of that group to compare the use of more than one-way ANOVA(one-way ANOVA) and SNK-q test.P＜0.05 considered statistically significant.Results 1.The survival rate of fetal rat As the hypoxic time lasting to reduce the survival rate of animals.2.Fetal rat condition Fetal rat weights of all hypoxic group were significantly lower than the air-control group(p＜0.01),as the hypoxic time lasting to reduce the Fetal rat weight.Right ventricle/left ventricle+septum(RV/LV + S) were significantly higher than those air control group(p＜0.01),not as the hypoxic time lasting to increase the RV/LV+S.3.Fetal lung condition Fetal lung weight of hypoxic 2-day group is decreased significantly lower than the air control group(p＜0.01),as the hypoxic time lasting to reduce the Fetal lung weight.Fetal lung /fetal rat weight of hypoxic 2-day group is decreased significantly lower than the air control group(p＜0.01), as the hypoxic time lasting to increase the fetal lung/fetal rat weight.4. Placental tissue HE staining Air-control group has normal structure of placental villi;all hypoxic group and control group are not different with structure of placental villi,umbilical artery of all hypoxic group and control group has no difference.5.Fetal lung tissue HE staining Air-control group alveolar structure is regular,arrangement orderly and distribution of alveolar septal uniformity,no significant exudative in alveolar space.As the hypoxic time lasting,alveolar structure is not ruley, arrangement is not orderly and distribution of alveolar septal is not uniformity,significant exudative in alveolar space.Air-control group,the pulmonary artery wall is thinner,lumen is larger.The hypoxia 10-day group,the pulmonary artery wall become thicker and lumen become narrower.6.Radial alveolar count(RAC) RAC of hypoxic 2-day group is decreased significantly lower than the air control group(p＜0.01), as the hypoxic time lasting to reduce the RAC.7.Pulmonary vascular morphometry As the hypoxic time lasting pulmonary vascular morphometry changes,wall area/total area(WA%),wall thickness(um),wall thickness/pulmonary arterial diameter(WT%) are increased in hypoxic 10-day group(P＜0.01)Conclusion1.Successfully established rat model of intrauterine hypoxia.2.Intrauterine hypoxia results in fetal rat growth restriction,fetal pulmonary hypoplasia;and as the time of intrauterine hypoxia lasting, fetal growth restriction is severity.ChapterⅡMK-801 on the dystrophy of fetal rat lung development induced by intrauterine hypoxia of different daysObjective:determine expression of NMDARmRNA under nomal and intrauterine hypoxia,Observe NMDA receptor antagonist(MK-801)'s role of fetal rat development under intrauterine hypoxia.Methods:1.Preparation of animal model and experimental division.Animal model as the first chapter.Experimental grouping as follows:air-control group,air+MK-801 group,hypoxia-control group, hypoxia+MK-801 group.2.NMDARmRNA expression in fetal lung tissue To extract part of lung tissue's RNA including the air-control group,and hypoxia- control group,detect the NMDA1 and 2A,B,C,D receptor mRNA in Real Time-PCR assay(finished by Shanghai shanjin company).3.Generally condiction results As the first chapter.4. Histopathological examination of placental As the first chapter.5. Lung tissue results As the first chapter.6.Statistical analysis Test data indicated that in mean±standard,deviation of that group to compare the use of more than one-way ANOVA(one-way ANOVA) and SNK-q test.P＜0.05 considered statistically significant.Results:1.NMDAR expression in fetal lung tissue of the air-control group,the main NMDAR's expression in fetal rat lung tissue is NMDAR2D;as the hypoxia time lasting,the expression of the NMDAR's expression in fetal lung tissue increased.In hypoxic six-day guope the expression is peak(p＜0.01),in which main expression of the NMDAR2D;In hypoxic ten-day guope the expression is decreased,no difference with the air-control group.2.The survival rate of fetal rat MK-801 increases the survival rate of all hypoxic group(p＜0.01).3. Fetal rat condiction Adding MK-801,reduced the hypoxia-induced fetal rat weight loss(p＜0.01).The air + MK-801 group compared with the air-control group,fetal rat weight is lighter than the latter(p＜0.01).Adding MK-801,reduced the hypoxia-induced fetal rat RV/LV+S gain(p＜0.01). The air + MK-801 group compared with the air-control group,fetal rat RV/LV+S is higher than the latter(p＜0.01).4.Fetal lung tissue HE staining The hypoxia+MK-801 group compared with the hypoxia group,the number of alveolar are more,alveolar structure is regularer, arrangement is orderly,distribution of alveolar septal is uniformity,cell's exudation reduced in alveolar chamber.The air+MK-801 group compared with the air-control group,the number of alveolar are less, alveolar structure is irregularer,arrangement is not orderly,distribution of alveolar septal is not uniformity,cell's exudation increased in alveolar chamber.The hypoxia+MK-801 group compared with the hypoxia group,pulmonary artery does not change obviously.The air+MK-801 group compared with the air-control group,no change in the pulmonary artery.5.Radial alveolar count(RAC) Adding MK-801,reduced the hypoxia-induced fetal rat RAC loss(p＜0.01).The air+MK-801 group compared with the air-control group,RAC is less than the latter(p＜0.01). 6.Pulmonary vascular morphometry The hypoxia+MK-801 group compared with the hypoxia group,has an amendment in pulmonary vascular morphometry.The hypoxia ten-day+MK-801 group compared with the hypoxia ten-day group,the wall area/vascular area(WA%), wall thickness(um),wall thickness/pulmonary arterial artery(WT%) are decreased(p＜0.05).The air + MK-801 group compared with the air-control group,has no difference in the pulmonary vascular morphometry.Conclusion1.The first observation the expression of NMDAR in lung development of fetal rat under normal and intrauterine hypoxia 2.The first MK-801 display a protective effect on development of fetal rat under intrauterine hypoxia,focusing on MK -801' s protective effects of fetal lung tissue development3.MK-801 has toxic effects on development of fetal rat alone used...