本文選題：氙氣 + 缺氧缺血�。� 參考：《青島大學(xué)》2016年碩士論文

【摘要】：目的:通過檢測(cè)腦白質(zhì)損傷的新生大鼠CLIC4表達(dá)水平變化,探討氙氣對(duì)腦白質(zhì)損傷的神經(jīng)保護(hù)作用,為臨床應(yīng)用氙氣治療新生兒腦損傷提供理論基礎(chǔ)和實(shí)驗(yàn)依據(jù)。方法:制作新生大鼠腦白質(zhì)損傷模型:選取生后48-72小時(shí)的SD新生大鼠144只,隨機(jī)分為生理鹽水(NS)組(空白對(duì)照組)(n=24)、脂多糖(LPS)組(n=24)、缺氧缺血(HI)組(n=24)、脂多糖+缺氧缺血(LPS+HI)組(腦損傷對(duì)照組)(n=24),LPS組僅給予腹腔注射LPS 0.05mg/kg,HI組分離結(jié)扎右側(cè)頸總動(dòng)脈以及置于8%的氮氧混合氣中1小時(shí)進(jìn)行缺氧缺血處理,LPS+HI組給予腹腔注射LPS 0.05mg/kg,3小時(shí)后進(jìn)行缺氧缺血處理,建立腦白質(zhì)損傷模型。NS組僅給予腹腔注射等量生理鹽水,不結(jié)扎不低氧處理。氙氣干預(yù)處理:通過脂多糖聯(lián)合缺氧缺血建立腦白質(zhì)損傷模型后,根據(jù)50%氙氣吸入開始時(shí)間分為A組(缺氧缺血后即刻)(n=24)、B組(缺氧缺血后2小時(shí))(n=24),進(jìn)行氙氣吸入3小時(shí)。各組分別于干預(yù)處理后0小時(shí)、24小時(shí)、48小時(shí)、72小時(shí)隨機(jī)各選取6只進(jìn)行甲醛灌注斷頭取腦,予蘇木素-伊紅染色(HE)觀察病理變化,髓鞘堿性蛋白(MBP)免疫熒光染色,并通過RT-PCR檢測(cè)腦組織中CLIC4mRNA水平變化。結(jié)果:1.NS組、HI組、LPS組腦白質(zhì)染色清晰,結(jié)構(gòu)正常;LPS+HI組新生大鼠腦組織HE染色可見腦白質(zhì)染色淡、結(jié)構(gòu)疏松;神經(jīng)纖維走向紊亂,神經(jīng)細(xì)胞染色加深,體積縮小,核仁變小,突起減少;膠質(zhì)細(xì)胞數(shù)量增多,可見細(xì)胞核固縮、胞漿疏松、細(xì)胞皺縮等凋亡改變。氙氣干預(yù)組新生大鼠腦白質(zhì)細(xì)胞排列紊亂,可見部分核固縮,但較LPS+HI組減輕。2.腦組織免疫熒光染色,在72小時(shí),NS組、HI組和LPS組腦組織可見大量MBP陽(yáng)性細(xì)胞;LPS+HI組MBP陽(yáng)性細(xì)胞數(shù)較NS組明顯減少。氙氣干預(yù)組MBP陽(yáng)性細(xì)胞較NS組減少、較LPS+HI組增多。3.腦損傷對(duì)照組及氙氣干預(yù)組CLIC4mRNA的表達(dá)水平高于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),表明損傷后腦組織中CLIC4mRNA的表達(dá)水平升高;但氙氣干預(yù)組CLIC4mRNA的表達(dá)水平低于腦損傷對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。24小時(shí)、48小時(shí)、72小時(shí)腦損傷對(duì)照組新生大鼠腦組織中CLIC4mRNA表達(dá)水平較空白對(duì)照組升高,氙氣干預(yù)組新生大鼠腦組織CLIC4mRNA表達(dá)水平在24小時(shí)、48小時(shí)、72小時(shí)較腦損傷對(duì)照組下降,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。但同日齡新生大鼠氙氣干預(yù)亞組中,A組與B組新生大鼠腦組織中CLIC4mRNA表達(dá)水平無明顯變化,差異無統(tǒng)計(jì)學(xué)意義(P0.05),表明CLIC4mRNA表達(dá)水平的高低與進(jìn)行氙氣干預(yù)的時(shí)間點(diǎn)無關(guān)。結(jié)論:1.腹腔注射LPS可與缺氧缺血共同作用,導(dǎo)致新生大鼠腦白質(zhì)損傷。2.腦白質(zhì)損傷后可使腦組織中CLIC4基因表達(dá)水平升高,說明CLIC4可引起神經(jīng)細(xì)胞損傷。3.氙氣干預(yù)能夠下調(diào)CLIC4基因表達(dá)水平,提示氙氣可能對(duì)中樞神經(jīng)系統(tǒng)存在保護(hù)作用。4.CLIC4基因表達(dá)水平的高低與腦白質(zhì)損傷后氙氣干預(yù)的時(shí)間點(diǎn)無關(guān),本研究中尚未發(fā)現(xiàn)進(jìn)行氙氣干預(yù)的最佳時(shí)間點(diǎn),提示在腦損傷后2小時(shí)內(nèi)應(yīng)用氙氣均可發(fā)揮其神經(jīng)保護(hù)作用。
[Abstract]:Objective: To explore the neuroprotective effect of xenon on brain white matter injury by detecting the changes of CLIC4 expression level in neonatal rats with brain white matter injury, and to provide theoretical basis and experimental basis for clinical application of xenon to treat neonatal brain injury. Methods: the model of brain white matter injury in newborn rats was made: 144 neonatal rats were selected for 48-72 hours after birth. Randomly divided into normal saline (NS) group (blank control group) (n=24), lipopolysaccharide (LPS) group (n=24), hypoxic-ischemic (HI) group (n=24), lipopolysaccharide + hypoxic-ischemic (LPS+HI) group (n=24), LPS group only given LPS 0.05mg/kg intraperitoneal injection, ligation of the right common carotid artery in the HI group and 1 hours of oxygen deficiency in 8% nitrogen oxygen mixture. Blood processing, group LPS+HI was given an intraperitoneal injection of LPS 0.05mg/kg, after 3 hours of hypoxic and ischemic treatment, the brain white matter injury model was established in group.NS only by intraperitoneal injection of equal amount of saline, no ligation and no oxygen treatment. Xenon intervention treatment: after the establishment of the brain white matter injury model by lipopolysaccharide combined with hypoxia ischemia, it began with the onset of 50% xenon inhalation. Group A (n=24), group B (2 hours after hypoxic ischemia) (n=24), xenon inhalation for 3 hours. Each group was randomly selected for 0 hours, 24 hours, 48 hours, 72 hours and 6 were randomly selected to perform formaldehyde perfusion to take the brain, and to observe pathological changes with hematoxylin eosin staining (HE) and myelin basic protein (MBP) immunofluorescence The changes of CLIC4mRNA level in brain tissue were detected by RT-PCR. Results: the white matter in group 1.NS, group HI and LPS group was clearly stained and the structure was normal. HE staining of brain tissue in group LPS+HI of neonatal rats showed that white matter was dyed light, structure loose, nerve fiber trend disorder, nerve cell staining deepened, volume narrowed, nucleolus became smaller, protuberance reduced; The number of cells increased, and the apoptotic changes were seen in nuclear condensation, cytoplasm loosening, and cell shrinkage. The brain white matter cells of neonatal rats in xenon intervention group were arranged in disorder, and partial nucleus retraction was visible, but a large number of MBP positive cells in group NS, HI group and LPS group were seen in group NS, HI group and LPS group, and LPS+HI group MBP positive in the group of LPS+HI. The number of cells in the xenon group was significantly lower than that in the NS group. The MBP positive cells in the xenon group were less than that in the NS group. The expression level of CLIC4mRNA in the.3. brain injury control group and the xenon group was higher than that in the xenon group. The difference was statistically significant (P0.05), indicating that the level of CLIC4mRNA expression in the brain tissue was increased after the injury, but the xenon intervention group was in CLIC4mRNA. The expression level was lower than that of the brain injury control group, the difference was statistically significant (P0.05).24 hours, 48 hours, 72 hours brain injury control group brain tissue CLIC4mRNA expression level increased compared with the blank control group, xenon group neonatal rats brain tissue CLIC4mRNA expression level at 24 hours, 48 hours, 72 hours compared with brain injury control group decreased. The difference was statistically significant (P0.05). However, there was no significant change in the expression of CLIC4mRNA in the brain tissue of A and B group rats in the xenon group of the same day old neonatal rats. The difference was not statistically significant (P0.05), indicating that the level of CLIC4mRNA expression was not related to the time point of xenon drying. Conclusion: 1. the intraperitoneal injection of LPS can be associated with hypoxia. The joint action of ischemia leads to the damage of white matter in neonatal rats with white matter damage, which can increase the level of CLIC4 gene expression in the brain tissue, indicating that CLIC4 can cause nerve cell damage and.3. xenon intervention can down regulate the expression of CLIC4 gene, suggesting that xenon may have a protective effect on the expression level of the.4.CLIC4 gene in the central nervous system. It is not related to the time point of xenon intervention after the injury of brain white matter. The best time point for xenon intervention has not been found in this study, suggesting that the application of xenon can play its neuroprotective effect within 2 hours after brain injury.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R742

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