【摘要】：背景 研究資料顯示宮內(nèi)窘迫會(huì)對(duì)大腦、腎臟、心臟、電解質(zhì)、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(yuǎn)、后果最嚴(yán)重的是對(duì)大腦的損傷，直接關(guān)系到新生兒的生存率和死亡率。目前宮內(nèi)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(jiàn)原因。尋求一種早期評(píng)估神經(jīng)系統(tǒng)發(fā)育狀態(tài)的方法迫在眉睫。近年相關(guān)研究指出促紅細(xì)胞生成素(erythropoietin, EPO)對(duì)神經(jīng)系統(tǒng)有保護(hù)作用，EPO可能對(duì)宮內(nèi)窘迫所致大腦損傷的預(yù)后有一定應(yīng)用價(jià)值。 目的 通過(guò)觀(guān)察EPO及其受體(EPOR)在宮內(nèi)窘迫大鼠生后腦組織的動(dòng)態(tài)表達(dá)規(guī)律，進(jìn)而探討EPO及EPOR在預(yù)測(cè)宮內(nèi)窘迫所致腦損傷中的價(jià)值，并同時(shí)探索外源性EPO治療新生動(dòng)物缺氧缺血性腦損傷的最佳時(shí)間窗。 方法 （1）采用結(jié)扎足月妊娠待產(chǎn)母鼠雙側(cè)子宮動(dòng)脈10分鐘，之后行剖宮產(chǎn)術(shù)的方法制作新生大鼠宮內(nèi)窘迫的模型，并以此作為實(shí)驗(yàn)組；未結(jié)扎雙側(cè)子宮動(dòng)脈的剖宮產(chǎn)新生大鼠作為對(duì)照組；（2）利用病理學(xué)技術(shù)，觀(guān)察實(shí)驗(yàn)組新生大鼠生后不同時(shí)間點(diǎn)（0h、2h、6h、12h、24h、48h、3d和7d）腦組織形態(tài)學(xué)方面的變化；以對(duì)照組新生鼠72h時(shí)點(diǎn)處的腦組織切片為參照物，對(duì)這兩組病理圖片進(jìn)行對(duì)比分析；（3）通過(guò)RT-PCR技術(shù)，研究新生大鼠生后不同時(shí)間點(diǎn)腦組織mRNA(EPO、EPOR)的表達(dá)情況；（4）利用Western blot實(shí)驗(yàn)技術(shù)，研究新生大鼠生后不同時(shí)間點(diǎn)腦組織EPO及EPOR的動(dòng)態(tài)表達(dá)情況；（5）利用ELISA實(shí)驗(yàn)技術(shù)檢測(cè)血清EPO的表達(dá)水平。實(shí)驗(yàn)所得數(shù)據(jù)用SPSS10.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果 與對(duì)照組相比，實(shí)驗(yàn)組新生大鼠腦組織中EPO蛋白和mRNA的表達(dá)在2h、6h、12h，3個(gè)時(shí)點(diǎn)明顯增強(qiáng)，差異有統(tǒng)計(jì)學(xué)意義（均P0.05）；EPOR蛋白和mRNA的表達(dá)2h、6h、12h、24h、48h、3d，，6個(gè)時(shí)點(diǎn)間均明顯增加，差異有統(tǒng)計(jì)學(xué)意義（均P0.05）。新生大鼠血清EPO水平在生后2h實(shí)驗(yàn)組高于對(duì)照組，差異有統(tǒng)計(jì)學(xué)意義（t=2.52, P=0.02）。 結(jié)論 （1）宮內(nèi)窘迫新生大鼠生后腦組織EPO與EPOR在短期內(nèi)迅速增加，其中EPOR持續(xù)高表達(dá)，且持續(xù)時(shí)間較EPO長(zhǎng)。（2）外源性EPO治療宮內(nèi)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(duì)宮內(nèi)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(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R722.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 丁璐;吳本清;黃進(jìn)潔;劉智屏;陳麗;;促紅細(xì)胞生成素對(duì)新生大鼠高氧肺損傷細(xì)胞凋亡的影響[J];中國(guó)當(dāng)代兒科雜志;2010年07期

2 李菊香;安陽(yáng)市人民醫(yī)院圍產(chǎn)兒死因臨床分析[J];河南預(yù)防醫(yī)學(xué)雜志;2001年01期

3 韓小英,于榮,劉維靖;胎兒宮內(nèi)生長(zhǎng)受限相關(guān)臨床因素的分析[J];首都醫(yī)科大學(xué)學(xué)報(bào);2005年03期

4 張永紅;文新忠;徐吉光;陳廣迪;鄭茂華;楊剛;火雷鳴;;重組人促紅細(xì)胞生成素對(duì)顱腦損傷患者的神經(jīng)保護(hù)作用[J];中華神經(jīng)外科疾病研究雜志;2008年03期

5 趙方;張雙船;周于新;曾賓;劉筱萍;;重組人促紅細(xì)胞生成素對(duì)新生鼠缺氧性腸損傷的防治作用[J];實(shí)用兒科臨床雜志;2009年11期

6 陳曉晴;許華;羅小平;黎萍;;促紅細(xì)胞生成素對(duì)缺氧缺血性腦損傷新生大鼠腦組織存活素表達(dá)及神經(jīng)細(xì)胞凋亡的影響[J];實(shí)用兒科臨床雜志;2011年02期

7 薄濤,嚴(yán)超英,霍淑芳,李巍;結(jié)扎孕鼠雙側(cè)子宮動(dòng)脈復(fù)制圍產(chǎn)期缺氧缺血性腦損傷動(dòng)物模型[J];中風(fēng)與神經(jīng)疾病雜志;2001年01期

8 顏淑芹,楊淑萍,劉秀霞,劉麗敏,薄文學(xué);結(jié)扎大鼠子宮建立新生乳鼠腦缺氧缺血模型[J];中國(guó)比較醫(yī)學(xué)雜志;2004年06期

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