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機(jī)場(chǎng)噪聲暴露對(duì)大鼠腦海馬區(qū)神經(jīng)元凋亡與Fas蛋白表達(dá)的影響

發(fā)布時(shí)間:2018-11-26 10:16
【摘要】: 目的 研究機(jī)場(chǎng)噪聲暴露對(duì)大鼠腦海馬區(qū)神經(jīng)元凋亡與Fas蛋白表達(dá)的影響。為噪聲污染影大鼠中樞神經(jīng)系統(tǒng)的相關(guān)研究提供理論依據(jù)。 方法 采用浙江大學(xué)環(huán)境物理實(shí)驗(yàn)室噪聲暴露后大鼠共16只作為實(shí)驗(yàn)?zāi)P。分為四組:正常飼養(yǎng)60天及90天的大鼠為對(duì)照組N1、N2,模擬蕭山機(jī)場(chǎng)噪聲暴露60天及脫離暴露后再正常飼養(yǎng)30天為實(shí)驗(yàn)組N3、N4。心臟灌注后行鼠腦切片,行Fas免疫組織化學(xué)染色、原位細(xì)胞凋亡檢測(cè)(TUNEL法),用Image-Pro Plus6.0軟件分析細(xì)胞陽性表達(dá)的積分光密度;另外再在透射電子顯微鏡下觀察大鼠海馬區(qū)神經(jīng)元超微結(jié)構(gòu)的改變。 結(jié)果 N1、N2組海馬內(nèi)Fas染色陽性表達(dá)的積分光密度分別為:2.67±1.67×103和3.03±2.18×103,兩者比較無明顯差異(P0.05);N1、N2海馬內(nèi)TUNEL陽性表達(dá)的積分光密度分別為:44.10±1.67和51.75±10.90,兩者比較亦無明顯差異(P0.05)。N3、N4組海馬內(nèi)Fas染色陽性表達(dá)的積分光密度分別為:34.29±12.60×103和20.54±5.69×103,兩者比較無明顯差異(P0.05),但上述兩組各自與N1、N2相比Fas染色陽性表達(dá)均顯著增加(P0.05);N3、N4海馬內(nèi)TUNEL陽性表達(dá)的積分光密度分別為:96.43±20.48和156.56±22.37,兩者比較亦無明顯差異(P0.05),N3、N4兩組各自與N1、N2相比TUNEL陽性表達(dá)均顯著增加(P0.05)。 透射電鏡觀察實(shí)驗(yàn)組海馬區(qū)神經(jīng)元胞核染色質(zhì)濃聚,濃縮成塊,列成沿核膜收縮的新月體,部分?jǐn)嗔?形成核碎片;細(xì)胞質(zhì)內(nèi)線粒體增大、空泡化,細(xì)胞骨架紊亂;細(xì)胞膜微絨毛、細(xì)胞突起及細(xì)胞間連接消失;出現(xiàn)凋亡小體;突觸間隙模糊,染色質(zhì)聚合,突觸小泡不清晰。突觸小泡減少,突觸后膜腫脹,散在小血管周圍膠質(zhì)膜腫脹。 結(jié)論 長期噪聲暴露可引起大鼠海馬區(qū)神經(jīng)元凋亡及Fas蛋白表達(dá)的增加。凋亡相關(guān)蛋白Fas表達(dá)的改變可能參與調(diào)控了細(xì)胞凋亡的過程。
[Abstract]:Objective to study the effects of airport noise exposure on neuronal apoptosis and Fas protein expression in rat hippocampal area. It provides a theoretical basis for the study of the central nervous system in rats exposed to noise pollution. Methods Sixteen rats were used as experimental models after noise exposure in the laboratory of environmental physics of Zhejiang University. The rats were divided into four groups: the control group was N1N _ 2 for 60 days and 90 days for normal feeding, and the experimental group was N3N _ 4 for 60 days of simulated noise exposure at Xiaoshan airport and 30 days of normal feeding for 30 days after detachment. After heart perfusion, brain sections were made, Fas immunohistochemical staining and in situ apoptosis assay (TUNEL) were performed. The integral optical density of positive cells was analyzed by Image-Pro Plus6.0 software. In addition, the ultrastructure of hippocampal neurons in rats was observed under transmission electron microscope. Results the integral optical density of Fas positive expression was 2.67 鹵1.67 脳 10 ~ 3 and 3.03 鹵2.18 脳 10 ~ 3 in N _ 1N _ 2 group, respectively. There was no significant difference between them (P0.05). The integral optical density of the positive expression of TUNEL in the hippocampus of N _ 1N _ 2 was 44.10 鹵1.67 and 51.75 鹵10.90, respectively, and there was no significant difference between the two groups (P0.05). The integral optical density of positive expression of Fas in hippocampus of N4 group was 34.29 鹵12.60 脳 10 ~ 3 and 20.54 鹵5.69 脳 10 ~ 3, respectively. There was no significant difference between the two groups (P0.05). The positive expression of N2 was significantly higher than that of Fas staining (P0.05). The integral optical density of the positive expression of TUNEL in the hippocampus of N3ON4 was 96.43 鹵20.48 and 156.56 鹵22.37respectively, and there was no significant difference between the two groups (P0.05). The positive expression of TUNEL in N2 was significantly higher than that in TUNEL (P0.05). Transmission electron microscope (TEM) was used to observe the chromatin accumulation, condensation and formation of crescents constricting along the nuclear membrane in the hippocampal neurons of the experimental group, and the mitochondria in the cytoplasm were enlarged, vacuolated, and the cytoskeleton was disordered. Cell membrane microvilli, cell processes and intercellular junctions disappeared, apoptotic bodies appeared, synaptic spaces were blurred, chromatin aggregation and synaptic vesicles were not clear. Synaptic vesicles decreased, postsynaptic membranes swelled and colloid membranes scattered around small vessels swelled. Conclusion long term noise exposure can induce neuronal apoptosis and increase the expression of Fas protein in hippocampal area of rats. The expression of apoptosis-related protein Fas may be involved in the regulation of apoptosis.
【學(xué)位授予單位】:浙江大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2010
【分類號(hào)】:R363

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