Metabolomics of Aurantio-Obtusin-Induced Hepatotoxicity in Rats for Discovery of Potential Biomarkers

Abstract

Aurantio-obtusin is an anthraquinone derived from Cassia obtusifolia (cassiae semen). It is also used as a tool and a detection index for the identification of cassiae semen, as stipulated by the Chinese Pharmacopoeia. Anthraquinones, the main components in cassiae semen, have been reported to show hepatotoxicity. This study investigates the hepatotoxicity of aurantio-obtusin in male Sprague–Dawley rats. We randomly divided the animals into a blank control group and treated three test groups with different doses of aurantio-obtusin: Low dose (4 mg/kg), medium dose (40 mg/kg), and high dose (200 mg/kg). Each group was treated with aurantio-obtusin for 28 days, whereas the control group was administered an equal volume of 0.5% carboxymethyl cellulose sodium salt (CMC-Na) aqueous solution. Subsequently, we conducted biochemical, hematological, and pathological investigations and determined the weight of different organs. We used serum metabolomics to identify possible biomarkers related to hepatotoxicity. The low-dose group showed no significant liver injury, whereas the medium- and high-dose groups manifested obvious liver injury. Compared with the control group, the test groups showed an increase in alanine transaminase, aspartate transaminase, and alkaline phosphatase levels. The liver organ coefficient also significantly increased. Additionally, we found significant changes in the hematological indices. Metabolomics analysis showed that aurantio-obtusin induced 28 endogenous markers related to liver injury. Our data indicate that aurantio-obtusin induces hepatotoxicity in rat liver in a dose-dependent manner and is mediated by pathways involving bile acids, fatty acids, amino acids, and energy metabolism. In particular, changes in bile acid content during treatment with therapeutic agents containing aurantio-obtusin deserve increased attention.

Urine Metabolomics Study on Potential Hepatoxic Biomarkers Identification in Rats Induced by Aurantio-Obtusin

“…In our previous study, we reported that aurantio-obtusin could cause liver injury in rats from the perspective of serum metabolomics combined with biochemical, hematological, and pathological indicators. We also found metabolic alterations in liver injury, including metabolism of amino acids (e.g., vanylglycol, phosphopantothenoylcysteine, and dehydroalanine) and lipids (e.g., cholic acid, taurodeoxycholic acid, and phosphatidylcholines) in a serum metabolomics study ( Xu et al., 2019 ). At present, there is no metabolomics applied to the study of urinary metabolism after aurantio-obtusin administration.…”

Aurantio-Obtusin Attenuates Non-Alcoholic Fatty Liver Disease Through AMPK-Mediated Autophagy and Fatty Acid Oxidation Pathways

“…All animals were accepted 1 week of adaptive feeding before the experiment and sacrificed after 8-weeks treatment. For in vivo experiments, the doses of AO (5,10 and 15 mg/kg) were selected based on recent publications ( Xu et al, 2019 ; Guo et al, 2021 ). In chronic HFSW experiment (n = 6), mice were divided into five groups: 1) control group (chow diet); 2) NAFLD model group [HFSW, Western diet-42% Kcal from fat and 0.2% cholesterol (TD.88137, Harlan Laboratories, Inc., Indianapolis, IN, United States) plus with a high sugar solution ( d -fructose: 23.1 g/L and d -glucose: 18.9 g/L) in drinking water ( Li et al, 2021 )]; (3–5) HFSW diet with AO administration group.…”

Predicting the potential toxicity of 26 components in Cassiae semen using in silico and in vitro approaches

“… “N”=negative, “P”=positive, “-”=not found. * in vivo results cited from the literature ( Program, 2001 , Wang et al, 2009 , Xu et al, 2019 , Yang et al, 2019b ). …”

“…As shown in Table 3 , Nos. 2 , 3 , 5 , 8 , 10, and 11 were reported to be hepatotoxic in vivo ( Program, 2001 , Wang et al, 2009 , Xu et al, 2019 , Yang et al, 2019b ), while only No. 10 was known to be nephrotoxic ( Wang et al, 2009 ).…”