| 气相色谱质谱联用仪定量检测内源性大麻素花生四烯乙醇胺及2-花生四烯酸甘油在胆道恶性肿瘤中的表达规律 |
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| 引用本文： | 袁锡裕,谭炳炎,黄力.气相色谱质谱联用仪定量检测内源性大麻素花生四烯乙醇胺及2-花生四烯酸甘油在胆道恶性肿瘤中的表达规律[J].中华普通外科学文献（电子版）,2019,13(5):345-349. |
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| 作者姓名： | 袁锡裕 谭炳炎 黄力 |
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| 作者单位： | 1. 523059 东莞市人民医院普外科
2. 510080 广州，中山大学公共卫生学院营养学系
3. 510080 广州，中山大学附属第一医院胆胰外科 |
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| 基金项目： | 国家自然科学基金资助项目(81201919); 广东省自然科学基金资助项目(2017A030313495); 东莞市社会科技发展项目(2016105101269) |
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| 摘 要： | 目的探讨两种主要的内源性大麻素花生四烯乙醇胺（AEA）及2-花生四烯酸甘油（2-AG）在胆道恶性肿瘤（BTC）患者体液及肿瘤组织中的表达规律。
方法采集中山大学附属第一医院2013年12月至2014年9月收治的22例BTC患者及8例良性胆道疾病患者的外周血及胆汁，其中15例BTC患者同时收集癌巢组织及癌旁正常组织，利用氯仿/甲醇抽提法配合固相萃取，分离患者体液及组织样本中的脂质，以二甲基异丙基硅烷-咪唑对脂质进行硅烷化处理，气相色谱质谱联用仪测定AEA及2-AG的浓度。
结果与良性胆道疾病患者相比，BTC患者血浆中2-AG显著升高180.01（140.72~283.84）nmol/L vs 42.33（25.61~148.93）nmol/L，P<0.05]，AEA浓度降低但差异无统计学意义；胆汁中AEA浓度显著降低1.80（0.50~5.00）nmol/L vs 10.15（2.68~17.49）nmol/L，P<0.05]，2-AG浓度升高但差异无统计学意义。BTC癌巢组织与癌旁正常组织相比，AEA浓度显著降低22.01（16.55~53.61）pmol/g vs 58.68（25.36~68.97）pmol/g，P<0.05]，2-AG显著升高1.97（1.44~5.43）nmol/g vs 1.10（0.36~1.47）nmol/g，P<0.05]。
结论本研究建立了在BTC中定量检测AEA和2-AG两种内源性大麻素浓度的有效方法，测定AEA及2-AG在BTC患者体液及肿瘤组织中的异常表达，可作为BTC的辅助诊断。
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| 关 键 词： | 胆管肿瘤 内源性大麻素 气相色谱-质谱法 花生四烯乙醇胺 2-花生四烯酸甘油 |
| 收稿时间： | 2019-05-22 |
Quantification of endocannabinoid arachidonoyl ethanolamide and 2-arachidonoylglycerol by gas chromatography/mass spectrometry in biliary tract cancer |
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| Xiyu Yuan,Bingyan Tan,Li Huang.Quantification of endocannabinoid arachidonoyl ethanolamide and 2-arachidonoylglycerol by gas chromatography/mass spectrometry in biliary tract cancer[J].Chinese Journal of General Surgery(Electronic Version),2019,13(5):345-349. |
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| Authors: | Xiyu Yuan Bingyan Tan Li Huang |
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| Institution: | 1. Department of General Surgery, Dongguan People’s Hospital, Dongguan 523059, China
2. Department ofNutrition, School of Public Health, Sun Yat-sen University, Guangzhou 510080, China
3. Department of Pancreatobiliary Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China |
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| Abstract: | ObjectiveTo investigate the expression of endocannabinoid (EC), arachidonoyl ethanolamide (AEA) and 2-arachidonoylglycerol (2-AG) in biliary tract cancer (BTC) patients’ samples.
MethodsLipids of patients’ plasma and bile (22 BTC patients and 8 benign biliary disease patients) and of paired BTC nest and adjacent normal tissue (15 of the above 22 BTC patients) were extracted by the Bligh and Dyer method (chloroform containing internal standards and methanol were added to above samples) and solid-phase extraction. Derivatisation of the hydroxyl groups of ECs was performed with the silylating agent dimethylisopropylsilyl imidazole. Lipid samples were quantitated by gas chromatography/electron ionization (EI)-mass spectrometry.
ResultsLevel of 2-AG was upregulated in BTC patients’ plasma 180.01 (140.72-283.84) nmol/L vs 42.33 (25.61-148.93) nmol/L, P<0.05], while level of AEA appeared decreasing trend without significant difference. Level of AEA was lower in BTC patients’ bile 1.80 (0.50-5.00) nmol/Lvs 10.15 (2.68-17.49) nmol/L, P<0.05], while level of 2-AG appeared increasing trend without significant difference. Level of AEA was lower 22.01 (16.55-53.61) pmol/g vs 58.68 (25.36-68.97) pmol/g, P<0.05], while level of 2-AG was upregulated 1.97 (1.44-5.43) nmol/g vs 1.10 (0.36-1.47) nmol/g, P<0.05] in BTC cancer nest than in adjacent normal tissues.
ConclusionsThe present study develops a sufficient and reliable method for quantitative determination of AEA and 2-AG in BTC. The dysregulated AEA and 2-AG in body fluids and tumor tissues of patients with BTC can be used as assistant diagnosis of BTC. |
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| Keywords: | Bile duct neoplasms Endocannabinoid Gas chromatography-mass spectrometry Arachidonoyl ethanolamide 2-Arachidonoylglycerol |
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