葛根素通过抑制AHR/氧化应激途径改善酮康唑诱导的大鼠肝损伤研究

doi:10.3969/j.issn.1672-5069.2024.05.006

摘要/Abstract

摘要： 目的探讨葛根素保护酮康唑诱导的大鼠肝损伤的机制。方法将48只SD大鼠随机分为对照组、模型组、大、小剂量葛根素干预组(n=12),采用酮康唑灌胃造模或在造模的同时给予葛根素灌胃干预。采用RT-PCR法检测肝组织芳香烃受体(AHR)、细胞色素p450家族CYP1A1和CYP2E1 mRNA水平,采用免疫组化法检测肝组织AHR蛋白表达。结果模型组大鼠血清酶和氧化应激指标显著升高,提示造模成功,而葛根素干预显著降低了它们的水平(P<0.05);小剂量葛根素处理组肝组织谷胱甘肽(GSH)水平为(148.2±9.5)μM,显著高于模型组【(77.0±9.1)μM,P<0.05】,而氧化型谷胱甘肽(GSSG)水平为(84.7±9.3)μM,显著低于模型组【(131.4±13.4)μM,P<0.05】,大剂量葛根素处理组有类似的变化,只是作用更强;小剂量葛根素处理组肝组织AHR、CYP1A1和CYP2E1 mRNA相对水平分别为(28.4±3.3)、(23.7±1.8)和(9.0±1.5),均显著低于模型组【分别为(51.7±7.8)、(36.2±4.7)和(14.0±1.5),P<0.05】,大剂量葛根素组表现为作用更强;葛根素处理组动物肝组织AHR表达显著弱于模型组。结论葛根素可能通过抑制芳香烃受体介导的氧化应激反应改善酮康唑诱导的大鼠肝损伤。

关键词: 药物性肝损伤, 酮康唑, 葛根素, 芳香烃受体, 抗氧化, 大鼠

Abstract: Objective This experiment aimed at exploring the mechanism ofprotective role of puerarin on ketoconazole-induced liver injury in rats. Method 48 male SD rats were randomly divided into control, model, low-dose and high-dose of puerarin-intervened groups (n=12 in each).Model was established by oral ketoconazole gavage, and intervention was carried out by oral ketoconazole and low-dose and large-dose of puerarin gavage simultaneously. Hepatic tissue aromatic hydrocarbon receptors (AHR), cytochrome P450 1A1(CYP1A1), and CYP2E1 mRNA levelswere assayed by RT-PCR, and hepatic expression of AHR protein was detected immunohistochemically. Results The model of liver injury was successfully established as proved enzymologically and histopathologically, and the intervention of puerarin greatly improved liver injury; hepatic tissue GSH level in low-dose puerarin-intervened group was (148.2±9.5)μM, much higher than [(77.0±9.1)μM,P<0.05], while GSSG level was (84.7±9.3)μM, much lower than [(131.4±13.4)μM, P<0.05] in the model, and in large-dose of puerarin intervention group, the changes were even more obviously; relative hepatic loads of AHR, CYP1A1 and CYP2E1 mRNA in low-dose of puerarin intervention were (28.4±3.3), (23.7±1.8)and (9.0±1.5), all significantly lower than [(51.7±7.8),(36.2±4.7) and (14.0±1.5), respectively, P<0.05] in the model, and the changes were even more obvious in large-dose of puerarin intervention; AHR expression in liver tissues with puerarin intervention was obviously weaker as compared in the model. Conclusion Puerarin could ameliorate ketoconazole-induced liver injury in rats, which might be related to inhibition of oxidative stress pathway mediated by AHR.

Key words: Drug-induced liver injury, Ketoconazole, Puerarin, Aromatic hydrocarbon receptors, Antioxidant, Rats

周云松, 赵琦, 胡芳, 李叙颖, 崔峻松, 黄敬, 黄娜娜, 赵一鹏. 葛根素通过抑制AHR/氧化应激途径改善酮康唑诱导的大鼠肝损伤研究[J]. 实用肝脏病杂志, 2024, 27(5): 669-672.

Zhou Yunsong, Zhao Qi, Hu Fang, et al. Puerarin improves ketoconazole-induced liver injury in rats by inhibiting aromatic hydrocarbon receptors/oxidative stress pathway[J]. Journal of Practical Hepatology, 2024, 27(5): 669-672.

参考文献

[1] 中国医药生物技术协会药物性肝损伤防治技术专业委员会, 中华医学会肝病学分会药物性肝病学组. 中国药物性肝损伤诊治指南(2023年版). 中华肝脏病杂志, 2023,31(4):355-384.
[2] Pivonello R, Simeoli C, Di Paola N, et al. Cushing's disease: adrenal steroidogenesis inhibitors. Pituitary, 2022,25(5):726-732.
[3] Tresnanda RI, Pramod SV, Safriadi F. Ketoconazole for the treatment of docetaxel-naïve metastatic castration-Resistant prostate cancer (mCRPC): asystematic review. Asian Pac J Cancer Prev, 2021;22(10):3101-3107.
[4] Korashy HM, Shayeganpour A, Brocks DR,et al. Induction of cytochrome P450 1A1 by ketoconazole and itraconazole but not fluconazole in murine and human hepatoma cell lines. Toxicol Sci, 2007,97(1):32-43.
[5] Zhou YS, Peng C, Zhou ZM, et al. Ketoconazole pretreatment ameliorates carbon tetrachloride-induced acute liver injury in rats by suppressing inflammation and oxidative stress.J Toxicol Sci, 2019,44(6):405-414.
[6] 周步高, 邹勇, 王馨, 等. 葛根素对损伤后肝细胞再生障碍大鼠的肝保护作用评价. 中华中医药杂志, 2017,32(5):2046-2049.
[7] Long M, Krüger T, Ghisari M, et al. Effects of selected phytoestrogens and their mixtures on the function of the thyroid hormone and the aryl hydrocarbon receptor. Nutr Cancer, 2012,64(7):1008-1019.
[8] Greenblatt DJ, Mikus G. Ketoconazole and liver injury: a five-year update. Clin PharmacolDrug Dev, 2019,8(1):6-8.
[9] Macho LP, Center SA, Randolph JF, et al. Clinical, clinicopathologic, and hepatic histopathologic features associated with probable ketoconazole-induced liver injury in dogs: 15 cases (2015-2018). J Am Vet Med Assoc, 2020,256(11):1245-1256.
[10] Khoza S, Moyo I, Ncube D. Comparative hepatotoxicity of fluconazole, ketoconazole, itraconazole, terbinafine, and griseofulvin in rats.J Toxicol, 2017,2017:6746989.
[11] Periasamy S, Liu CT, Chien SP, et al. Daily sesame oil supplementation mitigates ketoconazole-induced oxidative stress-mediated apoptosis and hepatic injury.J Nutr Biochem,2016;37:67-75.
[12] Sondermann NC, Faßbender S, Hartung F, et al. Functions of the aryl hydrocarbon receptor (AHR) beyond the canonical AHR/ARNT signaling pathway. Biochem Pharmacol, 2023,208:115371.
[13] Li H, Gao YH, Song L, et al. Ginsenoside Rg1 protects mice against 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced liver injury by inhibiting CYP1A1 through the aryl hydrocarbon receptor. J Ethnopharmacol, 2022,294:115394.
[14] Rath SN, Jena L, Patri M. Understanding ligands driven mechanism of wild and mutant aryl hydrocarbon receptor in presence of phytochemicals combating Parkinson's disease: an in silico and in vivo study. J Biomol Struct Dyn, 2020,38(3):807-826.
[15] Yu W, Zhao J, Li W, et al. 2,3,5,4'-Tetrahydroxystilbene-2-O-β-d-glucoside alleviated the acute hepatotoxicity and DNA damage in diethylnitrosamine- contaminated mice. Life Sci, 2020,243:117274.
[16] Diaz-Aguirre V,Velez-Pardo C,Jimenez-Del-Rio M.Fructose sensitizes Jurkat cells oxidative stress-induced apoptosis via caspase-dependent and caspase-independent mechanisms.CellBiolInt,2016,40(11):1162-117.
[17] Daud M, Majeed W, Awan AM, et al. Antioxidant and hepatoprotective activities of acacia jacquemontii stem extract against High-fat and CCl4-induced liver injury in rat's Model. Endocr Metab Immune Disord Drug Targets, 2023,23(1):77-85.
[18] Daniela G, Dimitrios T, Graziano C, et al. Pitfalls in the analysis of the physiological antioxidant glutathione (GSH) and its disulfide (GSSG) in biological samples: An elephant in the room. J Chromatogr B Analyt Technol Biomed Life Sci, 2016,15(1019):21-28.
[19] Cai Y, Sun W, Zhang XX, et al. Osthole prevents acetaminophen-induced liver injury in mice. Acta Pharmacol Sin, 2018,39(1):74-84.
[20] M C Guerra, E Speroni, M Broccoli, et al. Comparison between Chinese medical herb pueraria lobata crude extract and its main isoflavone puerarin antioxidant properties and effects on rat liver CYP-catalysed drug metabolism. Lif Sci, 2000,67: 2997-3006.