宫内窘迫大鼠脑组织促红细胞生成素及其受体的表达及意义

发布时间：2018-10-15 20:41

【摘要】：背景研究资料显示宫内窘迫会对大脑、肾脏、心脏、电解质、胃肠、代谢、肺脏、肝脏、视网膜造成不同程度的损害，其中损伤后影响最深远、后果最严重的是对大脑的损伤，直接关系到新生儿的生存率和死亡率。目前宫内窘迫是胎儿围产期死亡及新生儿神经系统后遗症的常见原因。寻求一种早期评估神经系统发育状态的方法迫在眉睫。近年相关研究指出促红细胞生成素(erythropoietin, EPO)对神经系统有保护作用，EPO可能对宫内窘迫所致大脑损伤的预后有一定应用价值。目的通过观察EPO及其受体(EPOR)在宫内窘迫大鼠生后脑组织的动态表达规律，进而探讨EPO及EPOR在预测宫内窘迫所致脑损伤中的价值，并同时探索外源性EPO治疗新生动物缺氧缺血性脑损伤的最佳时间窗。方法（1）采用结扎足月妊娠待产母鼠双侧子宫动脉10分钟，之后行剖宫产术的方法制作新生大鼠宫内窘迫的模型，并以此作为实验组；未结扎双侧子宫动脉的剖宫产新生大鼠作为对照组；（2）利用病理学技术，观察实验组新生大鼠生后不同时间点（0h、2h、6h、12h、24h、48h、3d和7d）脑组织形态学方面的变化；以对照组新生鼠72h时点处的脑组织切片为参照物，对这两组病理图片进行对比分析；（3）通过RT-PCR技术，研究新生大鼠生后不同时间点脑组织mRNA(EPO、EPOR)的表达情况；（4）利用Western blot实验技术，研究新生大鼠生后不同时间点脑组织EPO及EPOR的动态表达情况；（5）利用ELISA实验技术检测血清EPO的表达水平。实验所得数据用SPSS10.0软件进行统计学分析。结果与对照组相比，实验组新生大鼠脑组织中EPO蛋白和mRNA的表达在2h、6h、12h，3个时点明显增强，差异有统计学意义（均P0.05）；EPOR蛋白和mRNA的表达2h、6h、12h、24h、48h、3d，，6个时点间均明显增加，差异有统计学意义（均P0.05）。新生大鼠血清EPO水平在生后2h实验组高于对照组，差异有统计学意义（t=2.52, P=0.02）。结论（1）宫内窘迫新生大鼠生后脑组织EPO与EPOR在短期内迅速增加，其中EPOR持续高表达，且持续时间较EPO长。（2）外源性EPO治疗宫内窘迫致新生儿脑损伤具有可行性。（3）EPO与EPOR水平的变化规律对宫内窘迫致新生儿脑损伤的诊断和治疗具有一定的意义。
[Abstract]:Background data show that intrauterine distress causes varying degrees of damage to the brain, kidney, heart, electrolyte, gastrointestinal tract, metabolism, lung, liver, and retina, with the most profound impact after the injury. The most serious result is brain damage, directly related to neonatal survival and mortality. At present, intrauterine distress is a common cause of fetal perinatal death and neonatal neurological sequelae. It is urgent to find an early method to evaluate the developmental state of nervous system. Recent studies have pointed out that erythropoietin (erythropoietin, EPO) has protective effect on nervous system and EPO may be useful in prognosis of brain injury caused by intrauterine distress. Objective to observe the dynamic expression of EPO and its receptor (EPOR) in postnatal brain tissue of rats with intrauterine distress, and to explore the value of EPO and EPOR in predicting brain injury induced by intrauterine distress. At the same time, the best time window of exogenous EPO for the treatment of hypoxic ischemic brain injury in newborn animals was explored. Methods (1) the intrauterine distress model of newborn rats was established by ligation of bilateral uterine arteries for 10 minutes and then cesarean section. The neonatal rats without bilateral uterine artery ligation were used as control group. (2) pathological technique was used to observe the changes of brain morphology at different time points (0 h ~ 2 h ~ 6 h ~ 12 h ~ 24 h ~ 48 h / d) in experimental group. The brain tissue sections at 72 h in the control group were taken as the reference, the pathological pictures of the two groups were compared and analyzed; (3) the expression of mRNA (EPO,EPOR) in brain tissue at different time points after birth was studied by RT-PCR technique; (4) the expression of mRNA (EPO,EPOR) in the brain at different time points after birth was studied by Western blot technique. To study the dynamic expression of EPO and EPOR in brain tissue of neonatal rats at different time points after birth. (5) to detect the expression of serum EPO by ELISA assay. The experimental data were analyzed by SPSS10.0 software. Results compared with the control group, the expression of EPO protein and mRNA in the brain of neonatal rats in the experimental group was significantly increased at 2 h, 6 h and 12 h, and the difference was statistically significant (P0.05). The expression of); EPOR protein and mRNA were significantly increased at 2 h, 12 h, 24 h, 48 h and 3 d, respectively. The difference was statistically significant (P0.05). The level of serum EPO in neonatal rats was significantly higher than that in control group at 2 h after birth (t0. 52, P0. 02). Conclusion (1) EPO and EPOR in postnatal brain tissue of neonatal rats with intrauterine distress increased rapidly in a short period of time, and the expression of EPOR continued to be high. The duration is longer than that of EPO. (2) it is feasible to treat neonatal brain injury caused by intrauterine distress with exogenous EPO. (3) the changes of EPO and EPOR levels have certain significance in the diagnosis and treatment of neonatal brain injury caused by intrauterine distress.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R722.1

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