本文關(guān)鍵詞： 敵敵畏 毒性 代謝組學(xué) 長期低劑量 超高效液相色譜/飛行時間串聯(lián)質(zhì)譜　出處：《哈爾濱醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：用代謝組學(xué)方法研究長期低劑量經(jīng)口攝入敵敵畏對大鼠的毒性作用及可能的作用機制，篩選敵敵畏早期暴露的敏感生物標志物，為評價敵敵畏長期低劑量暴露對人類機體的影響提供更為可靠的理論依據(jù)。 方法：雄性Wistar大鼠（180g－220g）按體重隨機分為四組：低劑量組（大鼠經(jīng)口攝入敵敵畏的最大無作用劑量即NOAEL，2.4mg/kg bw/d），中劑量（7.2mg/kg bw/d,3×NOAEL），高劑量組（21.6mg/kg bw/d,9×NOAEL）和對照組。敵敵畏按照上述劑量通過飲水給予大鼠，連續(xù)染毒24周，對照組大鼠直接給予飲用水。在實驗進行4周、8周、12周、16周、20周和24周時，采集大鼠的血液、尿液和組織樣本，進行生化指標檢測、組織病理學(xué)檢測，同時應(yīng)用超高效液相色譜－質(zhì)譜聯(lián)用儀對尿液和血漿進行代謝組學(xué)分析。 結(jié)果：血清生化指標顯示，與對照組比較，中、高劑量組的膽堿酯酶(ChE)在染毒4周后即明顯升高（p＜0.05），谷氨酸氨基轉(zhuǎn)移酶（ALT）、天門冬氨酸轉(zhuǎn)氨酶（AST）、尿素氮（BUN）和肌酐（Cr）的含量在染毒12周后明顯升高（p＜0.05）；高劑量組白蛋白(ALB)在染毒12周后含量亦明顯降低（p＜0.05）。低劑量組的這幾個指標在實驗過程中均沒有發(fā)現(xiàn)明顯變化。組織病理學(xué)檢測顯示，實驗進行12周時，中、高劑量組肝組織出現(xiàn)脂肪變性或是細胞壞死等異常改變，低劑量組和對照組在整個實驗過程中無異常變化。 尿液代謝組學(xué)研究在正、負離子模式下共發(fā)現(xiàn)12個代謝物的異常改變。敵敵畏的代謝產(chǎn)物鄰苯二甲酸二甲酯（DMP），僅出現(xiàn)在染毒組，且隨著染毒劑量的增高，強度增加。與對照組比較，除了DMP，低劑量組尿液中的這些代謝物與對照組相比沒有顯著差異（p＞0.05）。中劑量組中吲哚酚硫酸和硫酸雌酮在敵敵畏處理12周后強度增高（p＜0.05），乳糖醛酸在實驗20周后強度升高（p＜0.05），尿酸、辛二酸、古洛糖酸、尿素和肌酐在實驗8周后強度顯著降低（p＜0.05），檸檬酸實驗進行12周后強度降低（p＜0.05）；高劑量組中的吲哚酚硫酸實驗8周后強度增高（p＜0.05），硫酸雌酮和乳糖醛酸實驗12周后強度增強（p＜0.05），尿酸（負離子模式）、辛二酸和檸檬酸在染毒8周后強度降低（p＜0.05），尿酸（正離子模式）、尿素和肌酐在整個實驗的24周強度均降低（p＜0.05）。 血漿代謝組學(xué)分析顯示，，與對照組相比，正、負離子模式下共發(fā)現(xiàn)10個代謝物的異常改變。低劑量組LysoPE(16:0/0:0)實驗16周后強度升高（p＜0.05）；LysoPC(17:0/0:0)、脫氫鞘氨醇、C16脫氫鞘氨醇和C17脫氫鞘氨醇染毒16周后強度降低（p＜0.05）。中劑量組LysoPE(16:0/0:0)實驗16周后強度升高（p＜0.05）；花生四烯酸、LysoPC(16:0/0:0)、 LysoPC(0:0/18:0)和C17脫氫鞘氨醇在實驗12周后強度降低（p＜0.05）；LysoPC(15:0/0:0)和鞘氨醇染毒處理16周強度降低（p＜0.05）；LysoPC(17:0/0:0)、脫氫鞘氨醇、C16脫氫鞘氨醇整個實驗過程全部降低（p＜0.05）。高劑量組中除代謝物L(fēng)ysoPE(16:0/0:0)實驗12周后強度升高（p＜0.05）外，其余9個代謝物在實驗過程中6個時間點強度全部降低（p＜0.05）。 結(jié)論：DMP作為敵敵畏在尿液中的代謝產(chǎn)物，具有靈敏性和特異性，可以作為敵敵畏暴露的一個生物標志物；大鼠暴露于敵敵畏后，影響了糖類、脂類的代謝及機體的抗氧化系統(tǒng)；尿液代謝組學(xué)分析顯示，在最大無作用劑量（NOAEL）水平上，沒有發(fā)現(xiàn)敵敵畏對大鼠機體的異常影響；血漿代謝組學(xué)分析顯示一些脂類的異常改變與臟器損傷有關(guān)；代謝組學(xué)技術(shù)在研究敵敵畏長期低劑量暴露毒性方面有獨特的優(yōu)勢。
[Abstract]:Objective: toxicity using metabonomics method to study the long-term low dose oral intake of dichlorvos in rats and its possible mechanism, screening of sensitive biomarkers for early exposure to dichlorvos, evaluation of dichlorvos long-term low dose exposure effect on the human body to provide a more reliable theoretical basis.
Methods: male Wistar rats (180g 220g) were randomly divided into four groups: low dose group (maximum dichlorvos oral intake of rats had no effect dose NOAEL, 2.4mg/kg bw/d), middle dose (7.2mg/kg bw/d, 3 * NOAEL), high dose group (21.6mg/kg bw/d, 9 x NOAEL) and control group. According to the above dose by drinking dichlorvos was given to rats, after 24 weeks, the rats in the control group given direct drinking water. For 4 weeks, 8 weeks, 12 weeks, 16 weeks in the experiment, 20 weeks and 24 weeks, rats collected blood, urine and tissue samples for biochemical detection, histopathological analysis, instrument of urine and plasma metabonomics analysis combined with ultra performance liquid chromatography and by mass spectrometry.
Results: the serum biochemical index, compared with the control group, high dose group, cholinesterase (ChE) in rats after 4 weeks was significantly increased (P < 0.05), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and creatinine (Cr) content increased significantly in after 12 weeks (P < 0.05); high dose group albumin (ALB) content in after 12 weeks were significantly lower (P < 0.05). The index of low dose group during the experiment were found no significant changes. By histopathology, the experiment for 12 weeks and in liver tissue of high dose group showed fatty degeneration or abnormal cell necrosis, low dose group and the control group had no abnormal changes during the whole experiment.
The urine metabonomic study in positive and negative ion mode were found abnormal changes of 12 metabolites. Two metabolites of phthalic acid methyl ester (DMP), two had only appeared in the control group, and with the increased dose intensity increased. Compared with the control group, in addition to DMP, the low dose group in urine these metabolites compared with the control group had no significant difference (P > 0.05). The middle dose group in indoxyl sulfate and estrone sulfate in the treatment of dichlorvos intensity increased after 12 weeks (P < 0.05), lactobionic acid in the 20 week after strength increased (P < 0.05), uric acid, suberic acid, gulonic acid in the experiment, urea and creatinine after 8 weeks of strength decreased significantly (P < 0.05), citric acid experiment strength decreased after 12 weeks (P < 0.05); after 8 weeks of indoxyl sulfate experiment in the high dose group strength increased (P < 0.05), estrone sulfate and aldehyde lactobionic acid after 12 weeks of strength test enhanced (P < 0.05), urine Acid (negative ion mode), octanic acid and citric acid decreased after 8 weeks of exposure (P < 0.05), uric acid (positive ion mode), urea and creatinine decreased at 24 weeks of the whole experiment (P < 0.05).
Plasma metabonomics analysis showed that compared with the control group, positive and negative ion mode were found abnormal changes of 10 metabolites. Low dose of LysoPE group (16:0/0:0) after 16 weeks of experiment the strength increased (P < 0.05); LysoPC (17:0/0:0), C16 dehydrogenation of sphingosine, dehydrogenation of sphingosine and dehydrogenation of C17 sphingosine after 16 weeks after the intensity decreased (P < 0.05). The middle dose group LysoPE (16:0/0:0) after 16 weeks of experiment the strength increased (P < 0.05); four arachidonic acid, LysoPC (16:0/0:0), LysoPC (0:0/18:0) to reduce the strength and C17 in the 12 week after dehydrogenation of sphingosine (P < 0.05); LysoPC (15:0/0:0) and sphingosine after 16 week treatment intensity decreased (P < 0.05); LysoPC (17:0/0:0), C16 dehydrogenation of sphingosine, dehydrogenation of sphingosine throughout the experiment all decreased (P < 0.05). High dose group except metabolite LysoPE (16:0/0:0) after 12 weeks of experiment the strength increased (P < 0.05), the remaining 9 metabolites in experimental process in The intensity of the 6 time points was all decreased (P < 0.05).
Conclusion: DMP as a metabolite of dichlorvos in the urine, with high sensitivity and specificity, can be used as a biomarker of exposure to dichlorvos; rats were exposed to DDVP, affect carbohydrate metabolism and lipid, antioxidant system of the display; urine metabolic analysis, the maximum non effect dose (NOAEL) level, no abnormal dichlorvos effects on rat body; plasma metabonomics analysis showed some abnormal changes of lipid related organ damage; metabonomics technique in the study of dichlorvos exposure has the unique advantage of poison.

【學(xué)位授予單位】：哈爾濱醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R114

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相關(guān)期刊論文 前1條

1 廖艷;彭雙清;顏賢忠;張立實;;抗結(jié)核藥物異煙肼肝毒性時效關(guān)系的代謝組學(xué)[J];中國醫(yī)學(xué)科學(xué)院學(xué)報;2007年06期

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