Cell and rat serum, urine and tissue metabolomics analysis elucidates the key pathway changes associated with chronic nephropathy and reveals the mechanism of action of rhein

Li Wang; Xixi Yu; Hongju Li; Dahong He; Su Zeng; Zheng Xiang

doi:10.1186/s13020-023-00862-1

Cell and rat serum, urine and tissue metabolomics analysis elucidates the key pathway changes associated with chronic nephropathy and reveals the mechanism of action of rhein

Chin Med. 2023 Dec 1;18(1):158. doi: 10.1186/s13020-023-00862-1.

Authors

Li Wang^{1

2

3}, Xixi Yu⁴, Hongju Li⁵, Dahong He², Su Zeng⁶, Zheng Xiang^{7

8}

Affiliations

¹ Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, People's Republic of China.
² Medical School, Hangzhou City University, Hangzhou, 310015, China.
³ Key Laboratory of Novel Targets and Drug Study for Neural Repair of Zhejiang Province, School of Medicine, Hangzhou City University, Hangzhou, 310015, China.
⁴ School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, China.
⁵ College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, 310018, China.
⁶ Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, People's Republic of China. zengsu@zju.edu.cn.
⁷ Medical School, Hangzhou City University, Hangzhou, 310015, China. XZH0077@126.com.
⁸ Key Laboratory of Novel Targets and Drug Study for Neural Repair of Zhejiang Province, School of Medicine, Hangzhou City University, Hangzhou, 310015, China. XZH0077@126.com.

Abstract

Background: Rhein can significantly delay the progression of chronic nephropathy. However, its mechanism of action has not been adequately elaborated, which hinders its extensive clinical application. In this work, the effects of rhein on models of TGF-β-induced NRK-49F cellular fibrosis and rat renal ischemia-reperfusion fibrosis were evaluated using metabolomics and western blotting.

Methods: The metabolic profiles of NRK-49F cells and rat urine, serum, and kidney tissues in the control, model, and rhein groups were investigated using UPLC-QTOF-MS. The levels of p-P65, p-IKK, p-AKT, p-P38, p-JNK and AP-1 in NRK-49F cells were measured using western blotting and immunofluorescence methods. Molecular docking and network pharmacology methods were employed to explore the relationship between the potential targets of rhein and key proteins in the NF-κB and MAPK signaling pathways.

Results: Various potential metabolites, including sphingolipids, ceramides, phosphatidylcholine, and lysophosphatidylcholine,14-hydroxy-E4-neuroprostane E, and 5-HPETE, were present in the cell, tissue, urine, and serum samples; however, few metabolites matches exactly among the four type of biological samples. These differential metabolites can effectively differentiated between the control, model, and rhein groups. Pathway enrichment analysis of differential metabolites unveiled that sphingolipid metabolism, arachidonic acid metabolism, and glycerophospholipid metabolism were closely related to nephropathy. Phosphorylation levels of AKT, IKK, P65 and AP-1 in NRK-49F cells was reduced by rhein treatment. Network pharmacology and molecular docking showed that the potential targets of rhein might regulated the expression of MAPK and AKT in the NF-κB and MAPK signaling pathways.

Conclusion: In brief, rhein might delays the progression of chronic nephropathy via the metabolic pathways, NF-κB and MAPKs signaling pathways, which provides the foundation for its development and clinical application.

Keywords: Chronic kidney disease; Fibrosis; Metabolomics; Rhein; UPLC-QTOF-MS.

Grants and funding

81973696/National Natural Science Foundation of China