【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。更多还原

【Abstract】 Objective To investigate the expression of endocannabinoid(EC), arachidonoyl ethanolamide(AEA) and 2-arachidonoylglycerol(2-AG) in biliary tract cancer(BTC) patients’ samples. Methods Lipids 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. Results Level 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/L vs 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. Conclusions The 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.更多还原