Integrated spatial metabolomics and transcriptomics reveal Ly6C⁺ macrophage glutamine metabolism in pneumonia-mediated attenuation of aristolochic acid I nephrotoxicity

Abstract

Objective: In traditional medicine, Asari Radix et Rhizoma (Xi Xin) is used to effectively treat respiratory diseases. However, the therapeutic portion of Xi Xin contains trace quantities of aristolochic acid I (AAI), which raises safety concerns. Furthermore, no compelling laboratory evidence confirms its safety. AAI-induced extensive renal tubular necrosis and inflammatory cell infiltration occurred primarily in the cortex and outer medulla. Accordingly, we examined the changes in metabolites within the aforementioned areas and thoroughly investigated the interactions between these differential metabolites and immune cells.

Methods: We mapped the spatial distribution of the differential metabolites L-glutamic acid and glutamine in mouse kidneys and explored the underlying mechanisms using transcriptomics and flow cytometry, further validating these findings through co-culture experiments in vitro.

Results: Administering 1 mg/kg AAI daily for 7 days (approximately 200 times the pharmacopeial Xi Xin dose) did not induce detectable levels of carcinogenic 7-(deoxyadenosin-N⁶-yl)-aristolactam I (dA-ALI) in mouse kidneys. However, dA-ALI was detected on the day after the administration of 10 mg/kg AAI. Mice with lipopolysaccharide-induced pneumonia exhibit increased tolerance to AAI-mediated nephrotoxicity. Based on integrated spatial metabolomics and renal transcriptomic analyses, increased tolerance to AAI-mediated nephrotoxicity may be related to glutamine-mediated oxidative stress regulation mechanisms. During pneumonia, mouse kidneys exhibit both immune and metabolic stress responses. Ly6C⁺ macrophages convert L-glutamic acid into glutamine, thereby reducing reactive oxygen species (ROS) levels in the extracellular matrix. This process, which is regulated by the ITGA5 receptor in renal tubular epithelial cells, modulates the pAkt/pNrf2/NQO1 pathway and reduces AAI-induced kidney damage.

Conclusions: Collectively, our findings indicate that Xi Xin is safe at conventional clinical dosages, and its targeted use can further minimize potential risks.

Keywords

Aristolochic acid I / Kidney damage / Oxidative stress / Spatial metabolomics / Xi Xin safety

Conflict of interest statement

The authors declare no conflict of interest.

Funding

This study was supported by the Consulting Project Funds of the Chinese Academy of Engineering (2023-XZ-88, China), Key Program of the National Natural Science Foundation of China (81721002 and 82230118, China), and Key Program of the National Natural Science Foundation of China Regional Union Fund (U23A20519, China).

Author contributions

Chengxian Li contributed to investigation, writing—original draft; Yinkang Wang contributed to data curation, visualization; Shuanglin Qin contributed to methodology, project administration; Xiaoyan Zhan contributed to writing—review & editing; Xu Zhao contributed to writing—review & editing; Mingxia Fang contributed to writing—review & editing; Jiaying Li contributed to validation; Ming Niu contributed to methodology, supervision; Zhaofang Bai contributed to conceptualization, funding acquisition; and Xiaohe Xiao contributed to conceptualization; project administration; funding acquisition. The manuscript has been read and approved by all authors, meets the journal’s criteria for authorship, and each author believes that the manuscript represents their honest work.

Ethical approval of studies and informed consent

All animal experiments were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals and were approved by the Fifth Medical Centre of the PLA General Hospital, Beijing, China (ethical approval no. IACUC-2021-0010).

Acknowledgments

None.

Data availability

All data generated or analyzed during this study are included in this published article.

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