Combined Metabolomics and Network Toxicology to Explore the Molecular Mechanism of Phytolacca acinose Roxb-Induced Hepatotoxicity in Zebrafish Larvae in Vivo.

Dan Cao,Meilin Chen,Ruichao Lin,Chongjun Zhao,Zhiqi Li,Jian Li,Haiyan Wang,Yangyu Jiang,Hanjun Ning,Qiqi Fan

doi:10.1155/2021/3303014

Abstract

Phytolacca acinosa Roxb (PAR), a traditional Chinese medicine, has been widely used as a diuretic drug for a long period of time for the treatment edema, swelling, and sores. However, it has been reported that PAR might induce hepatotoxicity, while the mechanisms of its toxic effect are still unclear. In this study, network toxicology and metabolomic technique were applied to explore PAR-induced hepatotoxicity on zebrafish larvae. We evaluated the effect of PAR on the ultrastructure and the function of the liver, predictive targets, and pathways in network toxicology, apoptosis of liver cells by PCR and western blot, and metabolic profile by GC-MS. PAR causes liver injury, abnormal liver function, and apoptosis in zebrafish. The level of arachidonic acid in endogenous metabolites treated with PAR was significantly increased, leading to oxidative stress in vivo. Excessive ROS further activated the p53 signal pathway and caspase family, which were obtained from KEGG enrichment analysis of network toxicology. The gene levels of caspase-3, caspase-8, and caspase-9 were significantly increased by RT-PCR, and the level of Caps3 protein was also significantly up-regulated through western blot. PAR exposure results in the liver function abnormal amino acid metabolism disturbance and motivates hepatocyte apoptosis, furthermore leading to liver injury.

Highlights

Phytolacca acinosa Roxb (PAR), namely, Shang Lu in Chinese, has a long history in China, which is widely used as a diuretic drug in treating various diseases such as edema, nephritis, ascites, swelling, and sores [1]
Over the course of the past decades, there has been a worldwide effort aimed at developing drugs from natural products to treat various health conditions and tend to make the Traditional Chinese Medicine PAR (TCM) as potential therapeutic options of worldwide importance. e reasons for this phenomenon may be related to a belief that herbal TCMs labeled as “natural” are always safe and of benefit to public health, improving physical fitness, extending lifespan, and treating various illnesses
The abundance of the reported liver injury cases induced by the TCM has attracted wide public and regulatory attention, especially related to hepatotoxicity, which can evolve into acute liver failure in most serious cases [22, 23]

Summary

Introduction

Phytolacca acinosa Roxb (PAR), namely, Shang Lu in Chinese, has a long history in China, which is widely used as a diuretic drug in treating various diseases such as edema, nephritis, ascites, swelling, and sores [1]. PAR is recorded in Chinese Pharmacopoeia and is famous for its multiple functions, such as antibacterial, antiinflammatory, antiviral, antitumor, and for enhancing immunity by modern pharmacological study [2]. The toxic performance of PAR cannot be ignored, which may have a significant effect on target organ damage [3]. A study showed that a large dose of esculentoside A, a main active ingredient in PAR, has toxicity on HK-2 (human renal tubular epithelial cells) and the mechanisms of toxicity are associated with cellular oxidative damage and cell apoptosis [4]. Another study found that esculentoside A reduced the viability of HL7702 (human normal liver cells), causing hepatotoxicity [5]. Until now, there have been few literatures on hepatotoxicity of PAR in vivo, and the mechanism of liver injury is still unclear

Methods

Results

Conclusion