Mechanism of Paeoniflorin in the Treatment of Bile Duct Ligation-Induced Cholestatic Liver Injury Using Integrated Metabolomics and Network Pharmacology.

Shizhang Wei,Jianxia Wen,Dan Wang,Xiao Ma,Ming Niu,Yanling Zhao,Haotian Li,Ruilin Wang,Tao Yang

doi:10.3389/fphar.2020.586806

Abstract

Paeoniflorin (PF) is the main active component of Paeonia lactiflora Pall., which is used in the treatment of severe cholestatic hepatitis. However, its biological mechanism in regulating bile acid metabolism and cholestatic liver injury has not been fully revealed. Our study aimed to reveal the mechanism of PF in the treatment of cholestatic liver injury in an in vivo metabolic environment using bioinformatics analysis. The serum of rats with bile duct ligation (BDL)-induced cholestatic liver injury treated with PF was analyzed by UHPLC-Q-TOF, and specific metabolites were screened using a metabolomics method. These specific metabolites were further analyzed by network pharmacology to identify the upstream signaling pathways and key protein targets. Finally, the key target proteins were verified by immunohistochemistry using cholestatic rat liver tissue. The serum ALT, AST, TBA, and TBIL levels, as well as the pathological state of the liver tissues, were significantly improved by PF. Twenty-five specific metabolites and 157 corresponding target proteins were screened for the treatment of cholestatic liver injury by PF. The “PF-target-metabolite” interaction network was constructed, and five protein targets (MAP2K1, MAPK1, ILBP, ABCB1, and LTA4H) that may regulate specific metabolites were obtained. The results of immunohistochemistry showed that PF improved the expression of these proteins. The integrated application of multiple bioinformatics methods revealed that PF plays a key role in the treatment of cholestatic liver injury by intervening in important targets related to bile acid metabolism and inflammation.

Highlights

Cholestasis, which is mainly characterized by abnormal bile acid metabolism, is caused by various factors, such as alcohol (Roth and Qin, 2019), viruses (Hou et al, 2017), drugs (Chatterjee and Annaert, 2018), and autoimmunity (Trivedi and Hirschfield, 2016)
To evaluate the effect of PF on cholestatic liver injury, we used bile duct ligation (BDL) rats to induce the pathological state of cholestasis, and different doses of PF (200 and 50 mg/kg) were administered by gastric administration
The results showed that PF regulated seven specific metabolites—leukotriene B4, taurocholic acid, biliverdin, taurochenodeoxycholic acid, glycocholic acid, lithocholic acid glycine conjugate, and sulfolithocholylglycine—through five targets (MAP2K1, mitogen-activated protein kinase 1 (MAPK1), ileal lipid-binding protein (ILBP), ABCB1, and leukotriene A-4 hydrolase (LTA4H)) in the interactive network

Summary

Introduction

Cholestasis, which is mainly characterized by abnormal bile acid metabolism (generation, secretion, and excretion), is caused by various factors, such as alcohol (Roth and Qin, 2019), viruses (Hou et al, 2017), drugs (Chatterjee and Annaert, 2018), and autoimmunity (Trivedi and Hirschfield, 2016). Bile cannot flow into the duodenum and enter the blood circulation in the pathological state, and toxic bile acids accumulate in the liver. Mechanism of Paeoniflorin Treating Cholestasis abnormalities and other signals are activated, leading to damage to the liver and bile duct cells. Continuous and severe cholestasis can progress to liver fibrosis, cirrhosis, hepatocellular carcinoma, and even liver failure (Pollock and Minuk, 2017). According to the pathological changes, cholestasis mainly manifests as hepatocellular and bile duct cholestasis. Primary biliary cholangitis (primary biliary cholangitis, PBC), drug-induced cholestasis, and primary sclerosing cholangitis (primary sclerosing cholangitis, PSC) are the most common presentations

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