本文選題：錳暴露　切入點：PARK2　出處：《遵義醫(yī)學院》2012年碩士論文

【摘要】：目的：探索錳暴露的早期效應標志物。方法：以錳接觸工人(男性)和SD大鼠(雄性)為實驗對象,采集工人空腹腔靜脈血10ml,儲存在4℃冰箱中備檢；SD大鼠24只隨機分為對照組(n=8)、低劑量組(n=8)和高劑量組(n=8)。通過腹腔注射染毒建立錳暴露模型,腹腔注射的錳溶液濃度為0mg/kg、5mg/kg,每周5次,共4周,實驗期間每次注射前測大鼠體重并記錄。染毒4周后斷頭處死大鼠,收集血液及腦組織。采用石墨爐原子吸收光譜法檢測全血中錳的含量；實時熒光定量聚合酶鏈反應(RT-PCR)法測全血中PARK2基因的表達情況及運用免疫組織化學技術檢測大鼠的腦組織中IARK2基因產(chǎn)物的含量。 結(jié)果：大鼠染錳1-3周內(nèi)體重迅速下降,錳暴露高劑量組與低劑量組平均體重都小于對照組,且隨著染錳濃度增大,平均體重呈逐漸降低趨勢(P0.05)。原子吸收法測定大鼠全血樣本中錳含量的結(jié)果顯示：與對照組相比較,錳含量在高劑量組與低劑量組增高,并且隨著染錳濃度增大呈逐漸增高趨勢(P0.05)；免疫組化結(jié)果顯示：PARK2基因在不同濃度的染錳大鼠腦組織各區(qū)域都有表達,主要位于神經(jīng)元胞漿中,錳暴露高劑量組與低劑量組各區(qū)域的PARK2基因表達產(chǎn)物含量都小于對照組,高劑量組各區(qū)域的PARK2基因的產(chǎn)物含量明顯小于低劑量組；實時熒光定量PCR結(jié)果顯示：大鼠在錳暴露4周后,高劑量組與低劑量的全血中ARK2基因的表達量小于對照組的表達量,且表達量的變化與染錳濃度成反比(P0.05),錳暴露工人的全血中PARK2基因水平低于對照組工人全血中的PARK2基因水平表達(P0.05)。 結(jié)論： 1.錳接觸可導致腦組織中PARK2基因表達產(chǎn)物降低,該機制可能是錳中毒機制之一。 2.血液中PARK2基因表達降低可能作為早期錳暴露的效應標志物。
[Abstract]:Objective: to explore the early effect markers of manganese exposure. Methods: workers exposed to manganese (male) and SD rats (male) were used as experimental objects. A total of 24 SD rats were randomly divided into two groups: control group (n = 8) and high dose group (n = 8). Manganese exposure model was established by intraperitoneal injection of manganese. The concentration of manganese solution injected intraperitoneally was 0 mg / kg 5 mg / kg, 5 times a week for 4 weeks. During the experiment, the weight of rats was measured and recorded before each injection. The rats were killed after 4 weeks of exposure. The blood and brain tissues were collected and the content of manganese in whole blood was determined by graphite furnace atomic absorption spectrometry (GFAAS). The expression of PARK2 gene in whole blood was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) and the content of IARK2 gene product in brain tissue of rats was detected by immunohistochemistry. Results: the body weight of rats decreased rapidly within 1-3 weeks after exposure to manganese. The average body weight of both high and low dose groups was lower than that of control group, and with the increase of manganese concentration, the weight of rats decreased rapidly. The results of atomic absorption spectrometry (AAS) for the determination of manganese content in whole blood samples of rats showed that compared with the control group, the manganese content in the high dose group and the low dose group was higher than that in the control group. With the increase of manganese concentration, the P0.05 gene was gradually increased, and the results of immunohistochemistry showed that the expression of 1% PARK2 gene was found in various regions of brain tissue of rats exposed to manganese at different concentrations, mainly in the cytoplasm of neurons. The content of PARK2 gene expression products in high dose group and low dose group was lower than that in control group, and the content of PARK2 gene in high dose group was significantly lower than that in low dose group. The results of real-time fluorescence quantitative PCR showed that the expression of ARK2 gene in high dose group and low dose whole blood was lower than that in control group after 4 weeks of manganese exposure. The level of PARK2 gene in the whole blood of the exposed workers was lower than that of the control group. The expression of PARK2 gene in the whole blood of the workers exposed to manganese was significantly lower than that of the control group. Conclusion:. 1. Manganese exposure may lead to the decrease of PARK2 gene expression in brain tissue, which may be one of the mechanisms of manganese poisoning. 2. The decrease of PARK2 gene expression in blood may be used as an effective marker of early manganese exposure.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R135

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