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

Abstract

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

CLC Number:

S831.5

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.

References

[1] HAKKOLA J, HUKKANEN J, TURPEINEN M, et al. Inhibition and induction of CYP enzymes in humans:an update[J]. Arch Toxicol, 2020, 94(11):3671-3722.
[2] ALNAIM L S, ALMALKI H M, ALMUTAIRI A M, et al. The prevalence of drug-drug interactions in cancer therapy and the clinical outcomes[J]. Life Sci, 2022, 310:121071.
[3] ZHANG Y J, GAO J, XU Y, et al. Investigation of cytochrome P450 inhibitory properties of deoxyshikonin, a bioactive compound from Lithospermum erythrorhizon Sieb. et Zucc[J/OL]. Phytother Res, 2022.[2023-07-03]https://onlinelibrary.wiley.com/doi/full/10.1002/ptr.7664.
[4] GUO J C, ZHU X H, BADAWY S, et al. Metabolism and mechanism of human cytochrome P450 enzyme 1A2[J]. Curr Drug Metab, 2021, 22(1):40-49.
[5] ZHOU S F, WANG B, YANG L P, et al. Structure, function, regulation and polymorphism and the clinical significance of human cytochrome P4501A2[J]. Drug Metab Rev, 2010, 42(2):268-354.
[6] 李新霞, 史荣梅, 刘睿婷. 大蒜主要化学成分及检测标准[J]. 新疆医科大学学报, 2020, 43(6):697-700.
LI X X, SHI R M, LIU R T. Main chemical components and testing standards of garlic[J]. Journal of Xinjiang Medical University, 2020, 43(6):697-700. (in Chinese)
[7] HUANG L R, JIA S F, WU R K, et al. The structure, antioxidant and antibacterial properties of thiol-modified soy protein isolate induced by allicin[J]. Food Chem, 2022, 396:133713.
[8] SÁNCHEZ-GLORIA J L, ARELLANO-BUENDIA A S, JUÁREZ-ROJAS J G, et al. Cellular mechanisms underlying the cardioprotective role of allicin on cardiovascular diseases[J]. Int J Mol Sci, 2022, 23(16):9082.
[9] XU S Y, LIAO Y X, WANG Q, et al. Current studies and potential future research directions on biological effects and related mechanisms of allicin[J]. Crit Rev Food Sci Nutr, 2022.[2023-07-03] https://www.tandfonline.com/doi/full/10.1080/10408398.2022.2049691.
[10] GONG H Z, LANG W Y, LAN H N, et al. Effects of laying breeder hens dietary β-carotene, curcumin, allicin, and sodium butyrate supplementation on the jejunal microbiota and immune response of their offspring chicks[J]. Poult Sci, 2020, 99(8):3807-3816.
[11] GONG H Z, WU M, LANG W Y, et al. Effects of laying breeder hens dietary β-carotene, curcumin, allicin, and sodium butyrate supplementation on the growth performance, immunity, and jejunum morphology of their offspring chicks[J]. Poult Sci, 2020, 99(1):151-162.
[12] HUANG R H, QIU X S, SHI F X, et al. Effects of dietary allicin on health and growth performance of weanling piglets and reduction in attractiveness of faeces to flies[J]. Animal, 2011, 5(2):304-311.
[13] 龚倩梅, 刘永仕, 袁何玲, 等. 大蒜辣素缓解CCl₄所致小鼠急性肝损伤的作用研究[J]. 南京农业大学学报, 2023, 46(3):530-537.
GONG Q M, LIU Y S, YUAN H L, et al. Effects of allicin on alleviating carbon tetrachloride-induced acute liver injury in mice[J]. Journal of Nanjing Agricultural University, 2023, 46(3):530-537. (in Chinese)
[14] LIANG Y, ZHANG J J, ZHANG Q B, et al. Release test of alliin/alliinase double-layer tablet by HPLC-Allicin determination[J]. J Pharm Anal, 2013, 3(3):187-192.
[15] 刘婉玉, BEKELE T G, 赵洪霞. 恩诺沙星在鲫鱼肝微粒体中的代谢及代谢关键酶[J]. 生态毒理学报, 2020, 15(3):64-70.
LIU W Y, BEKELE T G, ZHAO H X. Metabolism of enrofloxacin in liver microsomes of crucian carp (Caras-sius auratus) and its key enzymes in vitro[J]. Asian Journal of Ecotoxicology, 2020, 15(3):64-70. (in Chinese)
[16] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔC_T method[J]. Methods, 2001, 25(4):402-408.
[17] SZÜCS G, TAMÁSI V, LACZAY P, et al. Biochemical background of toxic interaction between tiamulin and monensin[J]. Chem Biol Interact, 2004, 147(2):151-161.
[18] RAVI P R, VATS R, KORA U R. Effect of ciprofloxacin and grapefruit juice on oral pharmacokinetics of riluzole in Wistar rats[J]. J Pharm Pharmacol, 2013, 65(3):337-344.
[19] FUHR U, ANDERS E M, MAHR G, et al. Inhibitory potency of quinolone antibacterial agents against cytochrome P450IA2 activity in vivo and in vitro[J]. Antimicrob Agents Chemother, 1992, 36(5):942-948.
[20] 陆苑, 潘洁, 谢玉敏, 等. 头花蓼水提取物对CYP450酶诱导和抑制作用研究[J]. 中国药理学通报, 2015, 31(8):1147-1152.
LU Y, PAN J, XIE Y M, et al. Inhibitory and inductive effects of Polygonum capitatum water extract on CYP450[J]. Chinese Pharmacological Bulletin, 2015, 31(8):1147-1152. (in Chinese)
[21] WANG H P, LI X X, LIU X Y, et al. Influence of pH, concentration and light on stability of allicin in garlic (Allium sativum L.) aqueous extract as measured by UPLC[J]. J Sci Food Agric, 2015, 95(9):1838-1844.
[22] MAITISHA G, AIMAITI M, AN Z C, et al. Allicin induces cell cycle arrest and apoptosis of breast cancer cells in vitro via modulating the p53 pathway[J]. Mol Biol Rep, 2021, 48(11):7261-7272.
[23] 常伟宇. 黄连素通过PXR信号通路调控CYP3A和P-gp的机制研究[D]. 广州:南方医科大学, 2016:1-96.
CHANG W Y. The mechanism research of berberine on regulation of CYP3A4 and P-gp via PXR signal pathways[D]. Guangzhou:Southern Medical University, 2016:1-96. (in Chinese)
[24] LI L, HAO H P, WANG Q, et al. Effects of short-term and long-term pretreatment of Schisandra lignans on regulating hepatic and intestinal CYP3A in rats[J]. Drug Metab Dispos, 2009, 37(12):2399-2407.
[25] 陈倩, 吴育晶, 程能能, 等. 五味子提取物对大鼠肝CYP3A酶的双重作用[J]. 药学学报, 2010, 45(9):1194-1198.
CHEN Q, WU Y J, CHENG N N, et al. Dual effects of extract of Schisandra chinensis baill on rat hepatic CYP3A[J]. Acta Pharmaceutica Sinica, 2010, 45(9):1194-1198. (in Chinese)
[26] ZHANG G Y, OU R L, LI F Y, et al. Regulation of drug-metabolizing enzymes and efflux transporters by Astragali radix decoction and its main bioactive compounds:Implication for clinical drug-drug interactions[J]. J Ethnopharmacol, 2016, 180:104-113.
[27] CHENG J, MA X C, GONZALEZ F J. Pregnane X receptor-and CYP3A4-humanized mouse models and their applications[J]. Br J Pharmacol, 2011, 163(3):461-468.
[28] WEI P, ZHANG J, EGAN-HAFLEY M, et al. The nuclear receptor CAR mediates specific xenobiotic induction of drug metabolism[J]. Nature, 2000, 407(6806):920-923.
[29] BAADER M, GNERRE C, STEGEMAN J J, et al. Transcriptional activation of cytochrome P450 CYP2C45 by drugs is mediated by the chicken xenobiotic receptor (CXR) interacting with a phenobarbital response enhancer unit[J]. J Biol Chem, 2002, 277(18):15647-15653.
[30] BU H Z, KNUTH K, MAGIS L, et al. High-throughput cytochrome P450(CYP) inhibition screening via a cassette probe-dosing strategy:V. Validation of a direct injection/on-line guard cartridge extraction-tandem mass spectrometry method for CYP1A2 inhibition assessment[J]. Eur J Pharm Sci, 2001, 12(4):447-452.
[31] 张玲玲. 离子载体类抗生素与氟喹诺酮类药物对AA肉鸡细胞色素P4501A和3A的影响及作用机制[D]. 南京:南京农业大学, 2010:1-153.
ZHANG L L. Effects and mechanism of ionophore antibiotics and fluoquinolones on cytochrome P4501A and 3A in AA broiler chicks[D]. Nanjing:Nanjing Agricultural University, 2010:1-153. (in Chinese)
[32] 范宰文, 马思琪, 徐爱军, 等. 3种中药成分对CYP2C9酶的影响[J]. 医药导报, 2019, 38(4):486-490.
FAN Z W, MA S Q, XU A J, et al. Effects of quercetin, paeoniflorin and oleanolic acid on CYP2C9 activities in vitro[J]. Herald of Medicine, 2019, 38(4):486-490. (in Chinese)
[33] 房绍英, 孙淑萌, 刘纯, 等. 多糖类组分对大鼠肝微粒体酶CYP1A2的体外活性抑制作用[J]. 中国现代应用药学, 2021, 38(24):3155-3159.
FANG S Y, SUN S M, LIU C, et al. Inhibitory effects of polysaccharides on the activity of CYP1A2 enzyme in rat liver microsomes in vitro[J]. Chinese Journal of Modern Applied Pharmacy, 2021, 38(24):3155-3159. (in Chinese)

Effects of Allicin on mRNA Expression of CYP1A2 and Its Enzyme Kinetics in Broilers Liver Tissue

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