二乙基亚硝胺诱导的肝癌模型大鼠尿液代谢组学研究
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摘要
目的应用核磁共振检测法(~1H-NMR)对大鼠尿液小分子代谢标志物及其参与的代谢通路进行分析。方法通过饮用质量浓度为0.1 mg·mL~(-1)的二乙基亚硝胺溶液(DEN)造肝癌大鼠模型,同时用饮用灭菌水的大鼠作为正常对照组,18周后采集尿样并进行核磁共振检测,在显微镜下对肝组织进行观察对比。结果模型组大鼠尿液中异亮氨酸、乳酸、丙氨酸、醋酸盐、糖蛋白、丙酮酸盐、琥珀酸盐、α-酮戊二酸、苯丙氨酸和甲酸盐10种代谢物的含量显著降低;而柠檬酸、二甲基亚硝酸、肌酸、牛磺酸和肌酸酐5种代谢产物的含量显著增加。采用MetaboAnalyst 3.0对差异性代谢物进行分析,发现共有4种代谢通路发生了明显变化。结论通过对大鼠尿液代谢物进行检测分析,为肝癌提供早期诊断依据。
Objective To analyze the metabolic markers of small molecules in urine and the metabolic pathways involved in the rat by hydrogen nuclear magnetic resonance(~1H-NMR) detection.Methods Rat models of hepatocellular carcinoma were established by drinking diethylnitrosamine(DEN) solution(0.1 mg·mL~(-1)),and rats were treated with sterilized water as normal control group.After 18 weeks,the urine samples were collected and analyzed by ~1H-NMR.The liver tissue was observed under microscope.Results In model group,10 metabolites,including isoleucine,lactic acid,alanine,acetate,glycoprotein,pyruvate,succinate,α-ketoglutaric acid,phenylalanine and formate were significantly decreased;the contents of citric acid,dimethylnitrosamine,creatine,taurine and creatinine were increased significantly.MetaboAnalyst 3.0 was used to analyze the different metabolites and 4 metabolic pathways were found changed significantly.Conclusion The detection and analysis of urinary metabolites in rats provide an early diagnostic basis for hepatocellular carcinoma.
Objective To analyze the metabolic markers of small molecules in urine and the metabolic pathways involved in the rat by hydrogen nuclear magnetic resonance(~1H-NMR) detection.Methods Rat models of hepatocellular carcinoma were established by drinking diethylnitrosamine(DEN) solution(0.1 mg·mL~(-1)),and rats were treated with sterilized water as normal control group.After 18 weeks,the urine samples were collected and analyzed by ~1H-NMR.The liver tissue was observed under microscope.Results In model group,10 metabolites,including isoleucine,lactic acid,alanine,acetate,glycoprotein,pyruvate,succinate,α-ketoglutaric acid,phenylalanine and formate were significantly decreased;the contents of citric acid,dimethylnitrosamine,creatine,taurine and creatinine were increased significantly.MetaboAnalyst 3.0 was used to analyze the different metabolites and 4 metabolic pathways were found changed significantly.Conclusion The detection and analysis of urinary metabolites in rats provide an early diagnostic basis for hepatocellular carcinoma.
引文
[1] 刘嘉,申恒巧,顾红梅,等.肝癌动物模型的构建及其应用现状[J].肿瘤药学,2013,3(1):13-16.
[2] 王倩,杨柱,龙奉玺,等.肝癌癌前病变动物模型研究进展[J].湖南中医杂志,2017,33(8):212-214.
[3] 练明建,刘树业.代谢组学在肝细胞肝癌标志物中的研究进展[J].临床检验杂志,2014,32(8):600-602.
[4] 罗艳,焦杨,邱莉,等.广西瑶药鸡爪风提取物抑制4种人肝癌细胞株体外增殖作用的研究[J].西北药学杂志,2014,29(4):376-380.
[5] 杨新宇,徐蓓蕾,沈芸.诱发性大鼠肝癌模型的建立[J].哈尔滨商业大学学报:自然科学版,2015,31(4):401-404,409.
[6] 王丹丹,张大方.诱发性大鼠肝癌模型的研究进展[J].吉林中医药,2006,26(10):75-76.
[7] 张海波,周荣耀.诱发性大鼠肝癌模型的研究现状与评价[J].医学综述,2002,8(6):320-322.
[8] Ding Y F,Wu Z H,Wei Y J,et al.Hepatic inflammation-fibrosis-cancer axis in the rat hepatocellular carcinoma induced by diethylnitrosamine[J].J Cancer Res Clin Oncol,2017,143(5):821-834.
[9] 陈晓,李涵,陈曦,等.代谢组学技术及其在药学研究领域的应用[J].武汉轻工大学学报,2016,35(2):12-15.
[10] 夏凯,贺晓云,郝俊冉,等.基于核磁共振扫描(NMR)的赭曲霉毒素A毒性血浆代谢组学研究[J].农业生物技术学报,2013,21(12):1426-1433.
[11] 刘悦,赵亮,李燕,等.肝细胞性肝癌代谢组学研究进展[J].药学实践杂志,2013,31(2):81-85.
[12] 李彦东,吴琪.代谢组学技术在临床诊断中的研究进展[J].天津医药,2015,43(8):942-945.
[13] 何梦秀,陈芳艳,钟杨生,等.γ-氨基丁酸代谢酶的研究进展[J].安徽农业科学,2015,43(30):47-50.
[14] 杨泽伟,王龙海,朱莉,等.γ-氨基丁酸代谢旁路在植物响应逆境胁迫中的作用机制研究[J].生物技术进展,2014,4(2):77-84.
[15] 殷海松,张仁宽,常燕钢,等.巴氏醋杆菌TCA循环代谢对醋酸发酵的影响[J].食品科学,2017,38(2):82-86.
[16] 李斌,黄艳,邵晨,等.铜离子急性胁迫对虎纹蛙肝脏中三羧酸循环及自由基代谢的影响[J].水生生物学报,2015,39(6):1160-1168.
[17] 程红焱,宋松泉.种子萌发过程中贮藏油脂的动员[J].云南植物研究,2007,29(1):67-73.
[18] 李尽贺,张涛,王明富,等.五味子纳米微粒水提液对衰老小鼠脑线粒体能量代谢的影响[J].中国老年学杂志,2008,28(7):667-668.
[19] 郭志新,John H McNeill.N-乙酰半胱氨酸对糖尿病大鼠肾脏NADPH 氧化酶表达的影响[J].中国糖尿病杂志,2008,16(9):544-547,553.
[20] 洪华,曾进胜,王莹,等.阿托伐他汀对大鼠缺血再灌注脑组织中NADPH 氧化酶源性超氧阴离子的抑制作用[J].中国病理生理杂志,2008,24(7):1345-1350.
[21] 苏兰鸿,孙林冲,高平.肿瘤的氨基酸代谢变化及其意义[J].中国生化药物杂志,2016,36(9):6-10.
[22] 芦佳.苯丙氨酸生物合成通路多酶基因协同表达研究[D].呼和浩特:内蒙古农业大学,2011.
[23] 王晓华,苏海翔.苯丙氨酸解氨酶抑制肝癌细胞生长的实验研究[J].肿瘤,1996,16(2):99-100.
[2] 王倩,杨柱,龙奉玺,等.肝癌癌前病变动物模型研究进展[J].湖南中医杂志,2017,33(8):212-214.
[3] 练明建,刘树业.代谢组学在肝细胞肝癌标志物中的研究进展[J].临床检验杂志,2014,32(8):600-602.
[4] 罗艳,焦杨,邱莉,等.广西瑶药鸡爪风提取物抑制4种人肝癌细胞株体外增殖作用的研究[J].西北药学杂志,2014,29(4):376-380.
[5] 杨新宇,徐蓓蕾,沈芸.诱发性大鼠肝癌模型的建立[J].哈尔滨商业大学学报:自然科学版,2015,31(4):401-404,409.
[6] 王丹丹,张大方.诱发性大鼠肝癌模型的研究进展[J].吉林中医药,2006,26(10):75-76.
[7] 张海波,周荣耀.诱发性大鼠肝癌模型的研究现状与评价[J].医学综述,2002,8(6):320-322.
[8] Ding Y F,Wu Z H,Wei Y J,et al.Hepatic inflammation-fibrosis-cancer axis in the rat hepatocellular carcinoma induced by diethylnitrosamine[J].J Cancer Res Clin Oncol,2017,143(5):821-834.
[9] 陈晓,李涵,陈曦,等.代谢组学技术及其在药学研究领域的应用[J].武汉轻工大学学报,2016,35(2):12-15.
[10] 夏凯,贺晓云,郝俊冉,等.基于核磁共振扫描(NMR)的赭曲霉毒素A毒性血浆代谢组学研究[J].农业生物技术学报,2013,21(12):1426-1433.
[11] 刘悦,赵亮,李燕,等.肝细胞性肝癌代谢组学研究进展[J].药学实践杂志,2013,31(2):81-85.
[12] 李彦东,吴琪.代谢组学技术在临床诊断中的研究进展[J].天津医药,2015,43(8):942-945.
[13] 何梦秀,陈芳艳,钟杨生,等.γ-氨基丁酸代谢酶的研究进展[J].安徽农业科学,2015,43(30):47-50.
[14] 杨泽伟,王龙海,朱莉,等.γ-氨基丁酸代谢旁路在植物响应逆境胁迫中的作用机制研究[J].生物技术进展,2014,4(2):77-84.
[15] 殷海松,张仁宽,常燕钢,等.巴氏醋杆菌TCA循环代谢对醋酸发酵的影响[J].食品科学,2017,38(2):82-86.
[16] 李斌,黄艳,邵晨,等.铜离子急性胁迫对虎纹蛙肝脏中三羧酸循环及自由基代谢的影响[J].水生生物学报,2015,39(6):1160-1168.
[17] 程红焱,宋松泉.种子萌发过程中贮藏油脂的动员[J].云南植物研究,2007,29(1):67-73.
[18] 李尽贺,张涛,王明富,等.五味子纳米微粒水提液对衰老小鼠脑线粒体能量代谢的影响[J].中国老年学杂志,2008,28(7):667-668.
[19] 郭志新,John H McNeill.N-乙酰半胱氨酸对糖尿病大鼠肾脏NADPH 氧化酶表达的影响[J].中国糖尿病杂志,2008,16(9):544-547,553.
[20] 洪华,曾进胜,王莹,等.阿托伐他汀对大鼠缺血再灌注脑组织中NADPH 氧化酶源性超氧阴离子的抑制作用[J].中国病理生理杂志,2008,24(7):1345-1350.
[21] 苏兰鸿,孙林冲,高平.肿瘤的氨基酸代谢变化及其意义[J].中国生化药物杂志,2016,36(9):6-10.
[22] 芦佳.苯丙氨酸生物合成通路多酶基因协同表达研究[D].呼和浩特:内蒙古农业大学,2011.
[23] 王晓华,苏海翔.苯丙氨酸解氨酶抑制肝癌细胞生长的实验研究[J].肿瘤,1996,16(2):99-100.