基于肝药酶CYP3A4、SULT2A1和UGT1A1探究京尼平的代谢解毒路径

doi:10.5246/jcps.2022.08.051

中国药学(英文版) ›› 2022, Vol. 31 ›› Issue (8): 608-621.DOI: 10.5246/jcps.2022.08.051

基于肝药酶CYP3A4、SULT2A1和UGT1A1探究京尼平的代谢解毒路径

张帆¹^,^#, 王晓慧²^,^#, Abdulaziz Ahmed A. Saad³, 席莉莉¹, 马小花¹, 石阿茜¹, 魏玉辉¹^,^*()

1. 兰州大学第一医院药剂科, 甘肃兰州 730000
2. 德州市产品质量标准计量研究院, 山东德州 253000
3. 兰州大学药学院, 甘肃兰州 730000

收稿日期:2022-02-11 修回日期:2022-03-28 接受日期:2022-04-26 出版日期:2022-09-03 发布日期:2022-09-03
通讯作者: 魏玉辉
作者简介:
+ Tel.: +86-931-8356510, E-mail: yhwei@lzu.edu.cn
基金资助:
National Natural Science Foundation of China (Grant No. 82174067, 81960646 and 82004080).

Hepatic CYP3A4, SULT2A1, and UGT1A1 synergistically mediate metabolic detoxification of genipin

Fan Zhang¹^,^#, Xiaohui Wang²^,^#, Abdulaziz Ahmed A. Saad³, Lili Xi¹, Xiaohua Ma¹, Axi Shi¹, Yuhui Wei¹^,^*()

1 Department of Pharmacy, The First Hospital of Lanzhou University, Lanzhou 730000, China
2 Dezhou Institute of Product Quality Standards and metrology, Dezhou 253000, China
3 School of pharmacy, Lanzhou University, Lanzhou 730000, China

Received:2022-02-11 Revised:2022-03-28 Accepted:2022-04-26 Online:2022-09-03 Published:2022-09-03
Contact: Yuhui Wei
About author:
# These authors contributed equally to this work.

摘要/Abstract

摘要：

栀子主要药效成分京尼平具有明显的肝毒性, 限制了京尼平以及栀子的应用和开发。本文主要以人源性HepaRG肝细胞为研究对象, 基于肝脏主要代谢解毒酶CYP3A4、SULT2A1和UGT1A1探究介导京尼平代谢解毒的具体路径。结果表明, CYP3A4、SULT2A1和UGT1A1协同介导了京尼平于肝脏的代谢解毒, 且CYP3A4是起决定因素的限速酶; 其最主要代谢解毒路径为: CYP3A4-SULT2A1/UGT1A1, 即: 京尼平进入肝细胞后, 首先经CYP3A4代谢为毒性减轻的Ⅰ相代谢产物, 再经Ⅱ相代谢酶SULT2A21和UGT1A1代谢为毒性进一步减轻的代谢产物。本研究的结果初步阐明了京尼平的肝代谢解毒路径, 为今后京尼平及栀子的合理使用及研发提供了依据, 也为将来京尼平以及中药栀子和含栀子的中药组方配伍解毒的研究奠定了基础。

关键词: 京尼平, 代谢解毒, CYP3A4, SULT2A1, UGT1A1, 栀子

Abstract:

In the present study, we aimed to ascertain the metabolic detoxification routes of genipin via CYP3A4, SULT2A1, and UGT1A1 in HepaRG cells. It was found that the hepatic CYP3A4, SULT2A1, and UGT1A1 synergistically mediated the metabolic detoxification of genipin, and the CYP3A4 was the limited enzyme. In detail, the pivotal detoxification pathway was CYP3A4-SULT2A1/UGT1A1, indicating that SULT2A1 and UGT1A1 further catalyzed the phase II detoxification metabolism followed by the genipin metabolization by CYP3A4 to the phase I metabolites with alleviated toxicity. Our findings provided valuable cues for future studies on the detoxification of genipin, even the compatibility detoxification of Zhi-zi. Moreover, these data facilitated the development and rational administration of genipin and Zhi-zi.

Key words: Genipin, Metabolic detoxification, CYP3A4, SULT2A1, UGT1A1, Gardenia jasminoides Ellis

Supporting:

张帆, 王晓慧, Abdulaziz Ahmed A. Saad, 席莉莉, 马小花, 石阿茜, 魏玉辉. 基于肝药酶CYP3A4、SULT2A1和UGT1A1探究京尼平的代谢解毒路径[J]. 中国药学(英文版), 2022, 31(8): 608-621.

Fan Zhang, Xiaohui Wang, Abdulaziz Ahmed A. Saad, Lili Xi, Xiaohua Ma, Axi Shi, Yuhui Wei. Hepatic CYP3A4, SULT2A1, and UGT1A1 synergistically mediate metabolic detoxification of genipin[J]. Journal of Chinese Pharmaceutical Sciences, 2022, 31(8): 608-621.

图/表 7

Figure 1. The respective effects of KTZ (CYP3A4 inhibitor) (A), GA (SULT2A1 inhibitor) (B), and Sor (UGT1A1 inhibitor) (C) on the HepaRG cell viabilities were analyzed by MTT (mean ± SD, n = 8 per group). *P < 0.05, **P < 0.01 indicated a statistically significant difference between inhibitor groups and the GP single group. GP: genipin; KTZ: ketoconazole; GA: glycyrrhetinic acid; Sor: sorafenib.

Figure 2. The respective effects of KTZ (CYP3A4 inhibitor) (A), GA (SULT2A1 inhibitor) (B), and Sor (UGT1A1 inhibitor) (C) on the LDH release rate (mean ± SD, n = 6 per group). *P < 0.05, **P < 0.01 indicated a statistically significant difference between inhibitor groups and the GP single group. GP: genipin; KTZ: ketoconazole; GA: glycyrrhetinic acid; Sor: sorafenib.

Figure 3. The effects of the specific inhibitors of CYP3A4, SULT2A1, and UGT1A1 on the intracellular concentrations of GP and their specific substrates (Tes, DHEA, and Bil), respectively. A, C, and E: The effect of KTZ, GA, and Sor on the intracellular concentrations of GP, respectively; B, D, and F: The impact of KTZ, GA, and Sor on the intracellular concentrations of Tes (CYP3A4 substrate), DHEA (SULT2A1 substrate), and Bil (UGT1A1 substrate), respectively. All data were expressed as mean ± SD (n = 3 per group). *P < 0.05, **P < 0.01 indicated a statistically significant difference between inhibitor groups and the GP single group or substrate groups. GP: genipin; KTZ: ketoconazole; Tes: testosterone; GA: glycyrrhetinic acid; DHEA: dehydroepiandrosterone; Sor: sorafenib; Bil: bilirubin.

Figure 4. The effects of the inhibitors combinations on the cell viability and LDH release rate. A and B: The effect of KTZ, GA, or/and Sor combinations on the cell viability (A) and LDH release rate (B); C and D: The cell viability ratio (C) and LDH release ratio (D) when KTZ (CYP3A4 inhibitor) was co-cultured as compared with GP single cultured. All data were expressed as mean ± SD (n = 8 per group for MTT analysis; n = 6 per group for LDH analysis). *P < 0.05, **P < 0.01 indicated a statistically significant difference between inhibitor groups and the GP single group or substrates groups. GP: genipin; KTZ: ketoconazole; GA: glycyrrhetinic acid; Sor: sorafenib.

Figure 5. The effects of the inhibitor combinations on the intracellular concentrations of GP (A) and its increased folds when KTZ (CYP3A4 inhibitor) was co-cultured compared with GP single cultured (B). All data were expressed as mean ± SD (n = 3 per group). *P < 0.05, **P < 0.01 with different colors indicated a statistically significant difference when the inhibitor groups with the corresponding colors were compared with the GP single groups. GP: genipin; KTZ: ketoconazole; GA: glycyrrhetinic acid; Sor: sorafenib.

Figure 6. The effects of rifampicin (enzyme inducer) on the cell viability (A), LDH release rate (B), intracellular concentrations of GP (C), and enzyme protein expressions (D). All data were expressed as mean ± SD (n = 6 per group for cell viability and LDH release rate analysis; n = 3 per group for protein expressions and intracellular concentrations of GP analysis). *P < 0.05, **P < 0.01 indicated a statistically significant difference as compared with the GP single groups and control group, respectively. GP: genipin; Rif: rifampicin.

Figure 7. Schematic overview of the metabolic detoxification pathways of GP mediated by CYP3A4, SULT2A1, and UGT1A1 in HepaRG cells. The chemical constructions of the phase I and II metabolites are referenced in the published articles[18,28,29].

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基于肝药酶CYP3A4、SULT2A1和UGT1A1探究京尼平的代谢解毒路径

Hepatic CYP3A4, SULT2A1, and UGT1A1 synergistically mediate metabolic detoxification of genipin

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