【摘要】：目的：通過(guò)研究錳暴露對(duì)腦組織、血液、唾液中PARK2表達(dá)的影響,探尋錳暴露早期效應(yīng)標(biāo)志物。方法：實(shí)驗(yàn)分兩部分進(jìn)行。人群研究部分：選擇錳鐵合金廠(chǎng)三年以上工齡的爐前工人作為錳接觸組(37名),以勞動(dòng)強(qiáng)度相似的鐵廠(chǎng)爐前工人作為對(duì)照組(41名)。原子吸收法(AAS)檢測(cè)各組血液和唾液中錳濃度,實(shí)時(shí)定量反轉(zhuǎn)錄-聚合酶鏈反應(yīng)法(RT-PCR)檢測(cè)各組血液和唾液中PARK2的表達(dá)。動(dòng)物實(shí)驗(yàn)部分：24只雄性SD大鼠(體重220g±10g)隨機(jī)分為三組：對(duì)照組(Omg/kg Mn),低劑量組(1mg/kg Mn),高劑量組(5mg/kg Mn)。通過(guò)腹腔注射法染錳(對(duì)照組注射等量生理鹽水),連續(xù)染錳四周,第四周末取大鼠腦組織(皮質(zhì)、紋狀體、海馬、丘腦)。原子吸收法(AAS)檢測(cè)各組腦組織中錳含量,實(shí)時(shí)定量反轉(zhuǎn)錄-聚合酶鏈反應(yīng)法(RT-PCR)檢測(cè)各組腦組織中PARK2表達(dá),蛋白質(zhì)免疫印跡法(WB)檢測(cè)各組腦組織中Parkin蛋白含量。結(jié)果：人群研究結(jié)果顯示：錳接觸組工人血液和唾液中錳濃度明顯高于對(duì)照組(P0.01)；與對(duì)照組比較,PARK2在血液和唾液中表達(dá)下降(P0.05)。動(dòng)物實(shí)驗(yàn)結(jié)果顯示：高劑量組和低劑量組大鼠(皮質(zhì)、紋狀體、海馬、丘腦)中錳含量高于對(duì)照組(P0.05,P0.01)；與對(duì)照組比較,PARK2在高劑量組大鼠(皮質(zhì)、紋狀體、海馬、丘腦)和低劑量組大鼠(紋狀體、海馬)中表達(dá)下降(P0.05),但是在低劑量組皮質(zhì)、丘腦中表達(dá)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)；Parkin蛋白在高劑量組大鼠(皮質(zhì)、紋狀體、海馬、丘腦)和低劑量組大鼠(紋狀體、海馬)中含量下降(P0.05,P0.01),但是在低劑量組大鼠皮質(zhì)、丘腦中差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論：1.錳暴露可使腦組織(皮質(zhì)、紋狀體、海馬、丘腦)、血液、唾液PARK2表達(dá)下降,提示可能是錳致神經(jīng)毒性的機(jī)制之一；2.唾液口腔脫落細(xì)胞中PARK2的表達(dá)變化有作為錳暴露后早期效應(yīng)標(biāo)志物的可能。
[Abstract]:Objective: To study the effect of manganese exposure on the expression of PARK2 in brain, blood and saliva, and to explore the early effect marker of manganese exposure. Methods: The experiment was divided into two parts. The study part of the population: the former workers in the manganese-iron alloy factory for more than three years were selected as the manganese contact group (37), and the workers in the iron factory with similar labor intensity were used as the control group (41). The concentration of manganese in each group of blood and saliva was detected by AAS. The expression of PARK2 in blood and saliva was detected by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Animal experiment part:24 male SD rats (weighing 220 g/10 g) were randomly divided into three groups: control group (Omg/ kg Mn), low-dose group (1 mg/ kg Mn) and high-dose group (5 mg/ kg Mn). The brain (cortex, striatum, hippocampus, thalamus) of the rat brain (cortex, striatum, hippocampus, thalamus) was obtained by intraperitoneal injection of manganese (the same amount of normal saline was injected into the control group). The expression of parkin in each group of brain tissue was detected by the method of atomic absorption (AAS), and the content of parkin in each group was detected by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results: The results of the population study showed that the concentration of manganese in the blood and saliva of workers exposed to manganese was significantly higher than that in the control group (P0.01). In comparison with the control group, the expression of PARK2 in blood and saliva decreased (P0.05). The results of animal experiments showed that the content of manganese in high-dose group and low-dose group (cortex, striatum, hippocampus and thalamus) was higher than that in the control group (P0.05, P0.01); in comparison with the control group, PARK2 was in high-dose group (cortex, striatum, hippocampus, thalamus) and low-dose group (striatum, The expression of Parkin in the rat (cortex, striatum, hippocampus, thalamus) and low-dose group (striatum, hippocampus) of rats (cortex, striatum, hippocampus, thalamus) and low-dose group (P0.05, P0.01) was not significant (P0.05). However, there was no significant difference in the cortex and thalamus of the low-dose group (P0.05). Conclusion:1. Manganese exposure may decrease the expression of PARK2 in the brain (cortex, striatum, hippocampus, thalamus), blood, and saliva, suggesting that it may be one of the mechanisms of manganese-induced neurotoxicity;2. The expression of PARK2 in saliva-shedding cells has the potential to be a marker of early effect after exposure to manganese.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R135

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