【摘要】：背景 研究資料顯示宮內窘迫會對大腦、腎臟、心臟、電解質、胃腸、代謝、肺臟、肝臟、視網(wǎng)膜造成不同程度的損害，其中損傷后影響最深遠、后果最嚴重的是對大腦的損傷，直接關系到新生兒的生存率和死亡率。目前宮內窘迫是胎兒圍產(chǎn)期死亡及新生兒神經(jīng)系統(tǒng)后遺癥的常見原因。尋求一種早期評估神經(jīng)系統(tǒng)發(fā)育狀態(tài)的方法迫在眉睫。近年相關研究指出促紅細胞生成素(erythropoietin, EPO)對神經(jīng)系統(tǒng)有保護作用，EPO可能對宮內窘迫所致大腦損傷的預后有一定應用價值。 目的 通過觀察EPO及其受體(EPOR)在宮內窘迫大鼠生后腦組織的動態(tài)表達規(guī)律，進而探討EPO及EPOR在預測宮內窘迫所致腦損傷中的價值，并同時探索外源性EPO治療新生動物缺氧缺血性腦損傷的最佳時間窗。 方法 （1）采用結扎足月妊娠待產(chǎn)母鼠雙側子宮動脈10分鐘，之后行剖宮產(chǎn)術的方法制作新生大鼠宮內窘迫的模型，并以此作為實驗組；未結扎雙側子宮動脈的剖宮產(chǎn)新生大鼠作為對照組；（2）利用病理學技術，觀察實驗組新生大鼠生后不同時間點（0h、2h、6h、12h、24h、48h、3d和7d）腦組織形態(tài)學方面的變化；以對照組新生鼠72h時點處的腦組織切片為參照物，對這兩組病理圖片進行對比分析；（3）通過RT-PCR技術，研究新生大鼠生后不同時間點腦組織mRNA(EPO、EPOR)的表達情況；（4）利用Western blot實驗技術，研究新生大鼠生后不同時間點腦組織EPO及EPOR的動態(tài)表達情況；（5）利用ELISA實驗技術檢測血清EPO的表達水平。實驗所得數(shù)據(jù)用SPSS10.0軟件進行統(tǒng)計學分析。 結果 與對照組相比，實驗組新生大鼠腦組織中EPO蛋白和mRNA的表達在2h、6h、12h，3個時點明顯增強，差異有統(tǒng)計學意義（均P0.05）；EPOR蛋白和mRNA的表達2h、6h、12h、24h、48h、3d，，6個時點間均明顯增加，差異有統(tǒng)計學意義（均P0.05）。新生大鼠血清EPO水平在生后2h實驗組高于對照組，差異有統(tǒng)計學意義（t=2.52, P=0.02）。 結論 （1）宮內窘迫新生大鼠生后腦組織EPO與EPOR在短期內迅速增加，其中EPOR持續(xù)高表達，且持續(xù)時間較EPO長。（2）外源性EPO治療宮內窘迫致新生兒腦損傷具有可行性。（3）EPO與EPOR水平的變化規(guī)律對宮內窘迫致新生兒腦損傷的診斷和治療具有一定的意義。
[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.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R722.1

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