發(fā)布時(shí)間：2018-06-13 07:42

  本文選題：促紅細(xì)胞生成素 + 腦白質(zhì)��； 參考：《青島大學(xué)》2015年博士論文

【摘要】：目的孕鼠腹腔注射脂多糖(lipopolysaccharide,LPS)制作宮內(nèi)炎癥致新生鼠腦白質(zhì)病變動(dòng)物模型。對(duì)腦白質(zhì)病變新生鼠腹腔注射重組促紅細(xì)胞生成素(recombinant human erythropoietin,rh EPO)rh EPO,通過(guò)內(nèi)源性EPO/EPOR m RNA及CD68、GFAP、MBP的檢測(cè)和神經(jīng)行為學(xué)評(píng)分,探討rh EPO對(duì)新生大鼠腦白質(zhì)病變近期和遠(yuǎn)期的神經(jīng)保護(hù)作用,為臨床應(yīng)用rh EPO治療早產(chǎn)兒腦白質(zhì)病變提供依據(jù)。方法30只Wistar孕鼠(孕齡15天)隨機(jī)數(shù)字法分為實(shí)驗(yàn)組和對(duì)照組。實(shí)驗(yàn)組腹腔注射LPS 300μg/kg制作早產(chǎn)兒腦白質(zhì)病變動(dòng)物模型,對(duì)照組Wistar大鼠腹腔注射生理鹽水(Normal saline,NS)1m L/kg作為對(duì)照。新生大鼠出生后,實(shí)驗(yàn)組與對(duì)照組各選30只新生鼠4%多聚甲醛灌注取腦,左側(cè)腦組織進(jìn)行HE染色,明確腦白質(zhì)損傷情況;右側(cè)腦組織用于實(shí)時(shí)熒光定量RT-PCR法檢測(cè)胼胝體促紅細(xì)胞生成素(erythropoietin,EPO)及EPOR m RNA水平。實(shí)驗(yàn)組隨機(jī)數(shù)字法取60只新生鼠隨機(jī)分為L(zhǎng)PS+rh EPO組(n=30)和LPS+NS組(n=30),分別于出生后即刻一次性腹腔注射rh EPO(5000IU/kg)和生理鹽水(1m L/kg)。對(duì)照組隨機(jī)數(shù)字法取60只新生鼠隨機(jī)分為NS+rh EPO組(n=30)和NS+NS組(n=30),分別于出生后即刻腹腔注射rh EPO(5000IU/kg)和生理鹽水(1m L/kg)。各處理組分別于用藥后后第3天、第7天隨機(jī)取10只新生鼠,進(jìn)行腦組織HE染色,免疫熒光法進(jìn)行CD68、GFAP及MBP的檢測(cè)。出生后2周各處理組隨機(jī)選取10只大鼠進(jìn)行神經(jīng)行為學(xué)評(píng)分。結(jié)果1.孕15天Wistar大鼠腹腔注射LPS后所產(chǎn)新生鼠HE染色腦室周圍腦白質(zhì)染色淡染,結(jié)構(gòu)稀疏,呈網(wǎng)狀改變;對(duì)照組腦白質(zhì)組織結(jié)構(gòu)正常。2.腹腔注射LPS大鼠所產(chǎn)新生鼠胼胝體EPO m RNA及EPOR m RNA的表達(dá)明顯高于腹腔注射NS組(P0.01)。3.生后3天HE染色顯示LPS+rh EPO組新生大鼠腦室周圍腦白質(zhì)染色淡染,結(jié)構(gòu)稀疏,呈網(wǎng)狀改變;LPS+NS組新生大鼠腦白質(zhì)結(jié)構(gòu)稀疏,較LPS+rh EPO組加重。生后7天,HE染色顯示LPS+rh EPO組腦室周圍白質(zhì)結(jié)構(gòu)稀疏,染色較清晰;LPS+NS組腦白質(zhì)結(jié)構(gòu)稀疏,呈網(wǎng)狀改變,染色不清晰。NS+rh EPO組及NS+NS組生后3天及7天腦白質(zhì)組織結(jié)構(gòu)正常,染色清晰。4.生后3天,LPS+rh EPO組、NS+rh EPO組、NS+NS組CD68表達(dá)均較LPS+NS組顯著下降(F=7.456,P0.01),生后7d,LPS+rh EPO組、NS+rh EPO組、NS+NS組GFAP表達(dá)明顯低于LPS+NS組(F=5.121,P0.01),生后3天(F=2.628)、7天(F=1.425)各組間MBP表達(dá)水平差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。5.LPS+rh EPO組生后2周神經(jīng)行為學(xué)評(píng)分(曠場(chǎng)試驗(yàn)、懸吊試驗(yàn)、斜坡試驗(yàn)、拒俘試驗(yàn))與LPS+NS組、NS+rh EPO組及NS+NS組均無(wú)明顯差別(F值分別為2.09、0.60、0.11、0.37)。結(jié)論1.宮內(nèi)炎癥致腦白質(zhì)損傷新生大鼠內(nèi)源性EPO m RNA及EPOR m RNA的表達(dá)明顯增加。2.rh EPO可抑制腦白質(zhì)損傷新生鼠的小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞活化。3.rh EPO腹腔注射對(duì)腦白質(zhì)損傷新生大鼠的遠(yuǎn)期神經(jīng)行為改善作用尚不明確。
[Abstract]:Objective to establish a rat model of intracerebral leukopathy induced by intrauterine inflammation by intraperitoneal injection of lipopolysaccharide (LPS) in pregnant rats. Recombinant human erythropoietine rh EPO-rh EPOs were injected intraperitoneally to neonate rats with leukoencephalopathy. The neuroprotective effects of rh EPO on neonate white matter lesions were investigated by the detection of endogenous EPO-EPOR mRNA and CD68 / GFAPMA-MBP and the neurobehavioral score of rh EPO in the near and long term. To provide evidence for clinical application of rh EPO in the treatment of white matter lesions of preterm infants. Methods Thirty pregnant Wistar rats (gestational age 15 days) were randomly divided into experimental group and control group. The experimental group was injected intraperitoneally with LPS-300 渭 g/kg to make the animal model of white matter disease of premature infants, and the control group was treated with normal saline NS1 L / kg as control group. After birth, 30 neonatal rats were perfused with 4% paraformaldehyde and the left brain tissues were stained with HE to determine the white matter injury. The right brain tissue was used to detect the level of erythropoietin (EPO) and EPOR mRNA in corpus callosum by real-time quantitative RT-PCR. In the experimental group, 60 neonatal rats were randomly divided into LPS rh EPO group (n = 30) and LPS NS group (n = 30). In the control group, 60 neonatal rats were randomly divided into NS rh EPO group (n = 30) and NS group (n = 30). Ten newborn rats were randomly selected on the 3rd and 7th day after treatment. The brain tissues were stained with HE and CD68 GFAP and MBP were detected by immunofluorescence. At 2 weeks after birth, 10 rats were randomly selected for neurobehavioral score in each treatment group. Result 1. After intraperitoneal injection of LPS on the 15th day of gestation, the neonate rats were stained by HE with light staining of white matter around the ventricle, with sparse structure and reticular changes, while the white matter of control group was normal. 2. The expression of EPO mRNA and EPOR mRNA in corpus callosum of neonatal rats produced by intraperitoneal injection of LPS was significantly higher than that in NS group. On the 3rd day after birth, HE staining showed that the white matter around the ventricle of the neonatal rats in LPS rh EPO group was light stained, the structure was sparse, and the reticular changes were observed. The white matter structure of the newborn rats in the LPS NS group was sparse, which was more serious than that in the LPS rh EPO group. On the 7th day after birth, HE staining showed that the white matter structure around the ventricle was sparse in LPS rh EPO group, the white matter structure was sparse and reticular in LPS NS group, and the white matter structure was normal in NS rh EPO group and NS group 3 and 7 days after birth. The staining was clear. On the 3rd day after birth, the expression of CD68 in NS rh EPO group was significantly lower than that in LPS NS group. The expression of GFAP in NS group was significantly lower than that in LPS rh EPO group on 7 days after birth. The expression of GFAP in NS group was significantly lower than that in LPS group (F5. 121 P 0. 01). No statistical significance was found in P0.05. 5. The neurobehavioral score of 2 weeks after birth in LPS rh EPO group (open field test). There was no significant difference between the suspension test, slope test and captivity test) and the NS rh EPO group and NS group in LPS NS group. The F values of the suspension test, the slope test and the NS group were 2.09 鹵0.60 and 0.110.37, respectively. Conclusion 1. Increased expression of endogenous EPO mRNA and EPOR mRNA in neonatal rats induced by intrauterine inflammation. 2. Rh EPO inhibited activation of microglia and astrocytes in neonatal rats with white matter injury The long-term neurobehavioral improvement in neonatal rats is unclear.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R742

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本文編號(hào)：2013252