【摘要】：研究目的:觀察天然抗氧化劑白藜蘆醇對(duì)腦出血模型大鼠繼發(fā)性腦損傷的神經(jīng)保護(hù)作用,并探討其發(fā)揮神經(jīng)保護(hù)作用的分子生物學(xué)影響。研究方法:SD雄性大鼠隨機(jī)分組:假手術(shù)組、腦出血組和白藜蘆醇組。所有實(shí)驗(yàn)?zāi)Ｐ痛笫竺刻炀谕粫r(shí)間段腹腔給藥一次,并分別利用行為學(xué)檢測方法疲勞轉(zhuǎn)棒實(shí)驗(yàn)和曠場實(shí)驗(yàn)檢測大鼠在造模前和造模后的運(yùn)動(dòng)能力變化,以評(píng)估其運(yùn)動(dòng)能力的恢復(fù)情況。在一系列行為學(xué)檢測完畢之后,模型大鼠被統(tǒng)一處死,取大腦半球標(biāo)本并固定。海馬區(qū)神經(jīng)元損傷及神經(jīng)細(xì)胞凋亡情況用尼氏染色方法檢測。用免疫熒光染色法標(biāo)記Iba-1檢測大腦皮質(zhì)區(qū)腦組織小膠質(zhì)細(xì)胞激活情況。并在高倍熒光倒置顯微鏡下進(jìn)行組織形態(tài)學(xué)觀察,用高清數(shù)碼相機(jī)拍攝觀察區(qū)域作后期數(shù)據(jù)定量分析。所有數(shù)據(jù)用統(tǒng)計(jì)學(xué)軟件進(jìn)行統(tǒng)計(jì)分析并制作圖表。結(jié)果:白藜蘆醇組大鼠在行為學(xué)實(shí)驗(yàn)中表現(xiàn)要顯著強(qiáng)于腦出血組大鼠,且在2周左右差異最為明顯。在疲勞轉(zhuǎn)棒實(shí)驗(yàn)中,腦出血組與白藜蘆醇組大鼠的運(yùn)動(dòng)能力在術(shù)后1天沒有明顯差異。從術(shù)后1周開始,白藜蘆醇組大鼠運(yùn)動(dòng)能力開始逐漸恢復(fù),而腦出血大鼠則是2周之后才開始出現(xiàn)運(yùn)動(dòng)能力恢復(fù)的跡象。白藜蘆醇組大鼠在給藥2周后相比腦出血組大鼠有顯著差異。在曠場實(shí)驗(yàn)中,腦出血組和白藜蘆醇組大鼠的運(yùn)動(dòng)距離,平均速度以及旋轉(zhuǎn)次數(shù)三項(xiàng)指標(biāo),相比于假手術(shù)組大鼠在術(shù)后1天檢測時(shí)均明顯下降。但在給藥1周后,白藜蘆醇組大鼠的運(yùn)動(dòng)距離和平均速度兩項(xiàng)指標(biāo)相比于腦出血組大鼠有明顯的差異,從一定程度上體現(xiàn)了白藜蘆醇組大鼠的運(yùn)動(dòng)能力恢復(fù)情況。在尼氏染色結(jié)果中發(fā)現(xiàn),大鼠海馬區(qū)腦組織的神經(jīng)元數(shù)量在造模之后顯著下降,但在給藥后,海馬區(qū)腦組織的神經(jīng)損傷明顯減輕,存活的神經(jīng)元數(shù)量相比腦出血組大鼠有顯著差異。在小膠質(zhì)細(xì)胞免疫熒光的結(jié)果中發(fā)現(xiàn),腦出血組和白藜蘆醇組大鼠在造模成功后,小膠質(zhì)細(xì)胞被大量激活,引起強(qiáng)烈的神經(jīng)炎癥反應(yīng)。而給藥之后的大鼠其小膠質(zhì)細(xì)胞激活數(shù)顯著降低,大鼠腦出血繼發(fā)性腦損傷引起的炎癥反應(yīng)得到一定程度的緩解。結(jié)論:本研究結(jié)果表明白藜蘆醇可以明顯促進(jìn)腦出血大鼠腦損傷后運(yùn)動(dòng)能力的恢復(fù),減少血腫所致的海馬區(qū)尼氏小體數(shù)量的減少和神經(jīng)元損傷。白藜蘆醇也可以通過減少小膠質(zhì)細(xì)胞的激活,起到抗神經(jīng)炎癥反應(yīng)的效果,從而有效地緩解腦出血后繼發(fā)性腦損傷。意義:研究白藜蘆醇對(duì)腦出血大鼠運(yùn)動(dòng)能力和神經(jīng)功能損傷的影響,進(jìn)一步驗(yàn)證可能的生物學(xué)調(diào)控作用,進(jìn)而為下一步毒理學(xué)、臨床預(yù)實(shí)驗(yàn)以及可能的藥物開發(fā)方向提供實(shí)驗(yàn)理論基礎(chǔ)。
[Abstract]:Aim: to observe the neuroprotective effect of resveratrol on secondary brain injury in rats with cerebral hemorrhage and to explore the molecular biological effects of resveratrol on neuroprotection. Methods male SD rats were randomly divided into three groups: sham operation group, intracerebral hemorrhage group and resveratrol group. All the experimental model rats were administered intraperitoneally every day at the same time, and the changes of motor ability of rats before and after modeling were measured by using behavioral test method, fatigue rotation test and open field experiment respectively. To assess the recovery of its athletic ability. After a series of behavioral tests were completed, the model rats were uniformly killed, the hemispheric specimens were taken and fixed. Neuronal injury and neuronal apoptosis in hippocampus were detected by Nissl staining. The activation of microglia in cerebral cortex was detected by immunofluorescence labeling Iba-1. The histomorphology was observed under high power fluorescence inverted microscope, and the quantitative analysis of the later data was made by using high-definition digital camera. All data are analyzed by statistical software and charts are made. Results: the behavior of resveratrol group was significantly stronger than that of intracerebral hemorrhage group, and the most obvious difference was about 2 weeks. There was no significant difference in motor ability between intracerebral hemorrhage group and resveratrol group 1 day after operation. From 1 week after operation, the motor ability of resveratrol group began to recover gradually, while that of intracerebral hemorrhage rats showed signs of recovery only after 2 weeks. Resveratrol group had significant difference compared with intracerebral hemorrhage group 2 weeks after administration. In the open field experiment, the movement distance, average speed and rotation times of rats in intracerebral hemorrhage group and resveratrol group were significantly lower than those in sham operation group. But after one week of administration, the two indexes of movement distance and average speed in resveratrol group were significantly different from those in intracerebral hemorrhage group, which reflected the recovery of motor ability in resveratrol group to some extent. The results of Nissl staining showed that the number of neurons in rat hippocampal brain tissue decreased significantly after the model was made, but the neuronal damage in hippocampal brain tissue was significantly alleviated after administration of the drug. The number of surviving neurons was significantly different from that of ICH rats. The results of microglia immunofluorescence showed that the microglial cells were activated in large quantities after the successful establishment of the model in intracerebral hemorrhage group and resveratrol group, which caused a strong neuroinflammatory reaction. However, the activation of microglia in rats was significantly decreased after administration of the drug, and the inflammatory response induced by secondary cerebral injury in rats with intracerebral hemorrhage was alleviated to a certain extent. Conclusion: resveratrol can significantly promote the recovery of motor ability and decrease the number of Nissl corpuscles and neuronal damage induced by intracerebral hemorrhage in rats with intracerebral hemorrhage. Resveratrol can also reduce the activation of microglia, play an anti-inflammatory effect, so as to effectively alleviate the secondary brain injury after intracerebral hemorrhage. Significance: to study the effects of resveratrol on motor ability and nerve function injury in rats with intracerebral hemorrhage. Clinical pre-trial and possible drug development provide experimental theoretical basis.
【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】：2017
【分類號(hào)】：R743.34

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