Abstract
Aristolochic acids (AAs) are a family of natural compounds with AA I and AA II being known carcinogens, whose bioactivation causes DNA adducts formation. However, other congeners have rarely been investigated. This study aimed to investigate genotoxicity of AA IVa, which differs from AA I by a hydroxyl group, abundant in Aristolochiaceae plants. AA IVa reacted with 2'-deoxyadenosine (dA) and 2'-deoxyguanosine (dG) to form three dA and five dG adducts as identified by high-resolution mass spectrometry, among which two dA and three dG adducts were detected in reactions of AA IVa with calf thymus DNA (CT DNA). However, no DNA adducts were detected in the kidney, liver, and forestomach of orally dosed mice at 40 mg/kg/day for 2 days, and bone marrow micronucleus assay also yielded negative results. Pharmacokinetic analyses of metabolites in plasma indicated that AA IVa was mainly O-demethylated to produce a metabolite with two hydroxyl groups, probably facilitating its excretion. Meanwhile, no reduced metabolites were detected. The competitive reaction of AA I and AA IVa with CT DNA, with adducts levels varying with pH of reaction revealed that AA IVa was significantly less reactive than AA I, probably by hydroxyl deprotonation of AA IVa, which was explained by theoretical calculations for reaction barriers, energy levels of the molecular orbits, and charges at the reaction sites. In brief, although it could form DNA adducts in vitro, AA IVa was non-genotoxic in vivo, which was attributed to its low reactivity and biotransformation into an easily excreted metabolite rather than bioactivation.
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Acknowledgements
We appreciate Dr. Tianpei Xie and Dr. Yong Qian for their helpful discussions and constructive suggestions. We thank Shanghai Standard Technology Co., Ltd. for providing the AA I and AA IVa.
Funding
This study was supported by the grants from the National Natural Science Foundation of China (Grant No. 81873081).
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JW performed experiment; collected and analyzed data; wrote the initial draft and revised the manuscript; prepared and created the published work. YL conceived and supervised the project; designed methodology (biology studies); verified the replication and reproducibility of results; reviewed and edited the manuscript; funding acquisition. XZ supervised the project; designed methodology (analytical chemistry studies); verified the replication and reproducibility of results; reviewed and edited the manuscript. ZY performed high-resolution mass spectrometric data collection in our studies. RC, JX, and YC participated in the experimental operation. YC performed density functional theory calculations of the study.
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Wan, J., Chen, R., Yang, Z. et al. Aristolochic acid IVa forms DNA adducts in vitro but is non-genotoxic in vivo. Arch Toxicol 95, 2839–2850 (2021). https://doi.org/10.1007/s00204-021-03077-1
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DOI: https://doi.org/10.1007/s00204-021-03077-1