Toxicity of Tetradium ruticarpum: Subacute Toxicity Assessment and Metabolomic Identification of Relevant Biomarkers

Qiyuan Shan; Gang Tian; Xin Han; Hui Hui; Mai Yamamoto; Min Hao; Jingwei Wang; Kuilong Wang; Xianan Sang; Luping Qin; Guanqun Chen; Gang Cao

doi:10.3389/fphar.2022.803855

Toxicity of Tetradium ruticarpum: Subacute Toxicity Assessment and Metabolomic Identification of Relevant Biomarkers

Front Pharmacol. 2022 Feb 28:13:803855. doi: 10.3389/fphar.2022.803855. eCollection 2022.

Authors

Qiyuan Shan^{1

2}, Gang Tian¹, Xin Han¹, Hui Hui¹, Mai Yamamoto³, Min Hao¹, Jingwei Wang⁴, Kuilong Wang¹, Xianan Sang¹, Luping Qin¹, Guanqun Chen², Gang Cao¹

Affiliations

¹ School of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou, China.
² Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, Canada.
³ Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada.
⁴ The Public Platform of Medical Research Center, Academy of Chinese Medical Science, Zhejiang Chinese Medical University, Hangzhou, China.

Abstract

Tetradium ruticarpum (TR) is widely used in Asia to treat gastrointestinal disorders and pain. Stir-frying with licorice aqueous extract is a traditional processing procedure of TR formed in a long-term practice and performed before clinical application, and believed to reduce TR's toxicity. However, its toxicity and possible toxicity attenuation approach are yet to be well investigated. Subacute toxicity and metabolomics studies were conducted to help understand the toxicity of TR. The subacute toxicity assessment indicated that 3 fold of the recommended therapeutic dose of TR did not show obvious subacute toxicity in rats. Although an extremely high dose (i.e., 60 fold of the recommended dose) may cause toxicity in rats, it reversed to normal after 2 weeks of recovery. Hepatocellular injury was the major toxic phenotype of TR-induced liver damage, indicating as aspartate aminotransferase (AST) and liver index increasing, with histopathologic findings as local hepatocyte necrosis, focal inflammatory cell infiltration, slightly bile duct hyperplasia, and partial hepatocyte vacuolation. Moreover, we evaluated the impact of processing in toxicity. TR processed with licorice could effectively reduce drug-induced toxicity, which is a valuable step in TR pretreatment before clinical application. Metabolomics profiling revealed that primary bile acid biosynthesis, steroid biosynthesis, and arachidonic acid metabolism were mainly involved in profiling the toxicity metabolic regulatory network. The processing procedure could back-regulate these three pathways, and may be in an Aryl hydrocarbon Receptor (AhR) dependent manner to alleviate the metabolic perturbations induced by TR. 7α-hydroxycholesterol, calcitriol, and taurocholic acid were screened and validated as the toxicity biomarkers of TR for potential clinical translation. Overall, the extensive subacute toxicity evaluation and metabolomic analysis would not only expand knowledge of the toxicity mechanisms of TR, but also provide scientific insight of traditional processing theory, and support clinical rational use of TR.

Keywords: Tetradium ruticarpum; metabolomics; toxicity; toxicity attenuation; toxicity biomarkers.