大蒜辣素对肉鸡肝组织中CYP1A2 mRNA表达及其酶动力学的影响

doi:10.11843/j.issn.0366-6964.2023.09.029

摘要/Abstract

摘要： 大蒜辣素(allicin)具有较好的抗菌、抗氧化等活性,有望作为一种抗生素替代品用于畜禽养殖业,但其对肝药物代谢酶的影响尚不清楚。本研究旨在探究大蒜辣素对肉鸡CYP1A2酶活性、编码基因的mRNA表达和酶动力学的影响,为其在生产实践中的合理使用提供参考。90只1日龄AA肉鸡适应性饲养4 d后随机分为对照组(基础日粮组)、大蒜辣素低剂量组(40 mg·kg^-1)和高剂量组(80 mg·kg^-1),处理组每天2次、连续42 d于饲料中添加大蒜辣素。给药后第14、28 和42天时每组分别随机采集10只鸡肝组织,钙离子沉淀法提取肝微粒体,检测大蒜辣素对CYP1A2酶活性的影响;采用real time RT-PCR方法检测大蒜辣素对CYP1A2和核受体CXR mRNA表达的影响;进一步采用体外探针药物法测定了大蒜辣素对 CYP1A2酶的抑制动力学参数。添加不同浓度大蒜辣素第14和28天,肉鸡肝微粒体中CYP1A2酶活性分别显著增加为对照组的1.2倍和1.4倍(P<0.05);而在第42天时低剂量和高剂量大蒜辣素分别使CYP1A2酶活性极显著减少至对照组的80%和62%(P<0.001)。大蒜辣素处理组对CYP1A2和CXR mRNA表达量与酶活性的影响趋势基本一致。体外试验结果显示,大蒜辣素对CYP1A2的半数抑制浓度(IC₅₀)为18.87 μmol·L^-1,抑制常数(K_i)为10.89 μmol·L^-1,并对CYP1A2呈现为非竞争性抑制作用,且其对CYP1A2酶活性的抑制作用呈现一定的时间和浓度依赖性。大蒜辣素可对CYP1A2酶活性及其编码基因的mRNA表达均产生影响,且表现为双向作用。该研究结果提示,大蒜辣素长期应用会对肉鸡CYP1A2产生影响,临床应注意其介导的药物间相互作用,避免产生不良反应。

关键词: 大蒜辣素, 肝微粒体, 肉鸡, CYP1A2, 酶动力学

Abstract: Allicin has good antibacterial and antioxidant activities, and is expected to be used as a substitute for antibiotics in the livestock and poultry industry, however, its effect on hepatic drug enzyme (CYP) remains unknown. The study was to investigate the effects of allicin on CYP1A2 enzyme activity, mRNA expression of coding gene and enzyme kinetics in broilers, and to provide a further reference for its rational use in production practice. Ninety one-day-old Arbor Acres (AA) broilers were randomly divided into control group (basal diet group), low-dose allicin group (40 mg·kg^-1) and high-dose allicin group (80 mg·kg^-1) after 4 days of adaptive breeding. The treatment groups were fed with allicin twice a day for 42 consecutive days. On the 14th, 28th, and 42nd day after treatment of allicin, 10 chickens from each group were randomly selected, respectively and liver microsomes were extracted by calcium ion precipitation method to detect the effects of allicin on CYP1A2 enzyme activity and enzyme kinetics parameters. The mRNA expression of CYP1A2 and nuclear receptor encoding gene CXR were detected by real time RT-PCR. Allicin (40 mg·kg^-1, and 80 mg·kg^-1) significantly increased the production of acetaminophen, the metabolite phenacetin catalyzed by CYP1A2,on the 14th and 28the day after treatment by 1.2 times and 1.4 times of the control group, respectively (P<0.05), however, the production of acetaminophen on the 42nd day, were significantly decreased to 80% and 62% of the control group, respectively (P<0.001). The variation trend of mRNA expression of CYP1A2 and CXR was consistent with that of enzyme activity. The half maximal inhibitory concentration (IC₅₀) of allicin on CYP1A2 was 18.87 μmol·L^-1, and the inhibition constant (K_i) was 10.89 μmol·L^-1. Allicin showed a non-competitive inhibition effect on CYP1A2 with a time and concentration-dependent manner. Allicin can affect both CYP1A2 enzyme activity and its encoding gene mRNA expression with a bidirectional manner. The results suggest that allicin has an inhibitory effect on broiler CYP1A2 after long-term application, and clinical attention should be paid to avoid the adverse reactions caused by drug-drug interaction.

Key words: allicin, liver microsome, broiler, CYP1A2, kinetics of enzyme

中图分类号:

S831.5

袁何玲, 方词, 黄金虎, 王晓明, 肖文浚, 刘睿婷, 史荣梅, 王丽平. 大蒜辣素对肉鸡肝组织中CYP1A2 mRNA表达及其酶动力学的影响[J]. 畜牧兽医学报, 2023, 54(9): 3905-3915.

YUAN Heling, FANG Ci, HUANG Jinhu, WANG Xiaoming, XIAO Wenjun, LIU Ruiting, SHI Rongmei, WANG Liping. Effects of Allicin on mRNA Expression of CYP1A2 and Its Enzyme Kinetics in Broilers Liver Tissue[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3905-3915.

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