锰暴露早期效应标志物初探

发布时间：2018-03-27 14:07

本文选题：锰暴露　切入点：PARK2　出处：《遵义医学院》2012年硕士论文

【摘要】：目的：探索锰暴露的早期效应标志物。方法：以锰接触工人(男性)和SD大鼠(雄性)为实验对象,采集工人空腹腔静脉血10ml,储存在4℃冰箱中备检；SD大鼠24只随机分为对照组(n=8)、低剂量组(n=8)和高剂量组(n=8)。通过腹腔注射染毒建立锰暴露模型,腹腔注射的锰溶液浓度为0mg/kg、5mg/kg,每周5次,共4周,实验期间每次注射前测大鼠体重并记录。染毒4周后断头处死大鼠,收集血液及脑组织。采用石墨炉原子吸收光谱法检测全血中锰的含量；实时荧光定量聚合酶链反应(RT-PCR)法测全血中PARK2基因的表达情况及运用免疫组织化学技术检测大鼠的脑组织中IARK2基因产物的含量。结果：大鼠染锰1-3周内体重迅速下降,锰暴露高剂量组与低剂量组平均体重都小于对照组,且随着染锰浓度增大,平均体重呈逐渐降低趋势(P0.05)。原子吸收法测定大鼠全血样本中锰含量的结果显示：与对照组相比较,锰含量在高剂量组与低剂量组增高,并且随着染锰浓度增大呈逐渐增高趋势(P0.05)；免疫组化结果显示：PARK2基因在不同浓度的染锰大鼠脑组织各区域都有表达,主要位于神经元胞浆中,锰暴露高剂量组与低剂量组各区域的PARK2基因表达产物含量都小于对照组,高剂量组各区域的PARK2基因的产物含量明显小于低剂量组；实时荧光定量PCR结果显示：大鼠在锰暴露4周后,高剂量组与低剂量的全血中ARK2基因的表达量小于对照组的表达量,且表达量的变化与染锰浓度成反比(P0.05),锰暴露工人的全血中PARK2基因水平低于对照组工人全血中的PARK2基因水平表达(P0.05)。结论： 1.锰接触可导致脑组织中PARK2基因表达产物降低,该机制可能是锰中毒机制之一。 2.血液中PARK2基因表达降低可能作为早期锰暴露的效应标志物。
[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.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R135

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