Penthorum chinense Pursh alleviates ethanol-induced hepatic oxidative impairment in zebrafish via AMPK / p62 / Nrf2 / mTOR pathway

Ethnopharmacological relevance: In China, Penthorum chinense Pursh (PCP) is renowned for its effectiveness in “promoting blood circulation” and “removing blood stasis”. It can “relieve the liver” and its application in the field of liver protection, including viral hepatitis, alcoholic liver, liver fibrosis, has been known for hundreds of years.Aim of the study: Oxidative stress is widely believed to exert a key role in the pathophysiology of alcoholic liver disease (ALD). Therefore, antioxidant therapy reflects a reasonable strategy for the prevention and treatment of ALD. Hence, this study aimed to elucidate the mechanism of PCP in ethanol-induced liver injury.Methods: Treatment of liver-specific transgenic zebrafish larvae (lfabp: EGFP) at three days post-fertilization (3 dpf) with different concentrations of PCP (100, 50, 25 μg / mL) for 48 h was followed by soaking in 350 mmol / L ethanol for 32 h. Liver function and fat accumulation were identified by phenotypic indicators and biochemical kits. The related proteins and gene expression were further estimated by western blotting and quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). Finally, high performance liquid chromatography (HPLC) was adopted to analyze the chemical composition of PCP extract.Results: Firstly, PCP mediated alleviation of ethanol-induced steatosis and reduction of aspartate aminotransferase (AST), alanine transaminase (ALT), total cholesterol (TC) and triglyceride (TG) related indexes were evident. Dose-dependent decrease of intracellular reactive oxygen species (ROS) production, the activity of malondialdehyde (MDA) and increased the activity of glutathione (GSH), Superoxide dismutase (SOD) and catalase (CAT) in zebrafish substantiated the role of PCP in relieving oxidative stress. Furthermore, PCP induced downregulation of sequestosome 1 (p62 / SQSTM1, p62), Atg13 and Beclin 1 expression promoted autophagy. Meanwhile, PCP contributed to the hepatoprotective function by downregulating the expression of kelch-like ECH-associated protein 1 (Keap1) and upregulating the expression of nucleus factor-E2-related factor 2 (Nrf2), which activated cytoprotective related gene HO-1. Moreover, HPLC of PCP extract confirmed the presence of various polyphenols with potential antioxidant effects. Finally, PCP appeared to promote the activated protein kinase (AMPK) / p62 / Nrf2 / mTOR signaling pathways, which were related to oxidative stress and autophagy in zebrafish.Conclusion: This study claimed that by activating the AMPK / p62 / Nrf2 / mTOR signaling pathway, PCP could attenuate ethanol-induced liver injury in zebrafish.

Penthorum Chinense Pursh Protects Liver From Alcohol-induced Steatosis in Zebrafish by Mechanisms Including Inhibition of Oxidative Stress and Increase in Autophagy

BackgroundAs an ATP-gated ion channel, P2X7 receptor (P2X7R) affects lipid metabolism by activating the dangerous molecule ATP derived from liver cells caused by alcohol. Penthorum chinense Pursh (PCP), known as “shenxiancao”, plays a significant role in treating liver disease among Miao people. We first investigated whether liver protection mechanism of PCP mediated by P2X7R. MethodsFirst, treatment of zebrafish transgenic (fabp10: EGFP) larvae with different concentrations of PCP after 48 h at 3 dpf, then soaked in 350 mmol/L ethanol for 32 h. Subsequently the ameliorative effect of PCP in zebrafish with alcoholic hepatosteatosis was studied. In addition, gene expression related to lipid metabolism, oxidative stress, and autophagy was detected from the mRNA level by RT-qPCR and related proteins were measured by Western blot. Then, larvae were stimulated with ATP alone to explore whether PCP was the target of P2X7R.ResultsPCP significantly improved liver function and lipid deposition in zebrafish with alcoholic hepatosteatosis, and regulated the expression of SREBP1, CHREBP and FAS by elevating LKB1 and AMPK, thereby inhibiting the synthesis of fatty acids. Also, SIRT1 was suppressed in the model group, while PCP upregulated the expression. Inecreased expression of PPARα, decreased PPARγ, and CPT1 then promoted the oxidation of fatty acids. PCP dose-dependently decreased intracellular ROS production in zebrafish, then reduced MDA activity elevation and increased GSH, CAT and SOD levels. The specific mechanism may be realized by up-regulating the antioxidant pathway of Keap1/Nrf2 and down-regulating the autophagy pathway of PI3K/Akt/mTOR to regulate lipid metabolism. After ATP stimulation, P2X7R is further activated, which in turn regulated Keap1/Nrf2 and mTOR/PI3K/Akt mRNA expression, while PCP reversed these changes.ConclusionsPCP may be a target of P2X7R involvement in the regulation of this mechanism through up-regulation of the antioxidant pathway of Keap1/Nrf2 and down-regulation of the autophagic pathway of mTOR/ PI3K/Akt to regulate lipid metabolism, suggesting that blocking P2X7R is an emerging strategy for the therapy of ALD.

An Overview of the Mechanism of Penthorum chinense Pursh on Alcoholic Fatty Liver

Alcohol liver disease (ALD) caused by excessive alcohol consumption is a progressive disease, and alcohol fatty liver disease is the primary stage. Currently, there is no approved drug for its treatment. Abstinence is the best way to heal, but patients’ compliance is poor. Unlike other chronic diseases, alcohol fatty liver disease is not caused by nutritional deficiencies; it is caused by the molecular action of ingested alcohol and its metabolites. More and more studies have shown the potential of Penthorum chinense Pursh (PCP) in the clinical use of alcohol fatty liver treatment. The purpose of this paper is to reveal from the essence of PCP treatment of alcohol liver mechanism mainly by the ethanol dehydrogenase (ADH) and microsomal ethanol oxidation system-dependent cytochrome P4502E1 (CYP2E1) to exert antilipogenesis, antioxidant, anti-inflammatory, antiapoptotic, and autophagy effects, with special emphasis on its mechanisms related to SIRT1/AMPK, KEAP-1/Nrf2, and TLR4/NF-κB. Overall, data from the literature shows that PCP appears to be a promising hepatoprotective traditional Chinese medicine (TCM).

Genetic authentication of Eclipta prostrate (Asteraceae) from Penthorum chinense (Penthoraceae) by Sequence Characterized Amplified Region (SCAR) markers

Introduction: For the rapid and accurate genetic identification and authentication of living organisms, improved random amplified polymorphic DNA (RAPD) fragment based development of sequence-characterized amplified region (SCAR) markers is an important genetic technique. Objective: This study aimed to develop SCAR markers for perennial herb Eclipta prostrate (E. prostrate). Methods: Here the RAPD fragments by improved RAPD amplification with primers A11 and N-7 for E. prostrate were cloned into pGEX-T vector, and PCR amplification identified the positive clones. After the enzymatic digestion, they were sequenced with Sanger sequencing. Results: Two SCAR markers were developed, which were very specific to E. prostrate, not found in Penthorum chinense Pursh (P. chinense). The nucleotide sequence search by BLAST GenBank database showed that they are novel in E. prostrate, therefore they were deposited in Genbank with accession number KX671034, KX671035. The markers did not show any identity to other species. Conclusions: Thus, in this study two specific SCAR markers were developed for genetically distinguishing and identifying the plant species E. prostrate from herb P. chinense and others.

Kaempferol from Penthorum chinense Pursh suppresses HMGB1/TLR4/NF-κB signaling and NLRP3 inflammasome activation in acetaminophen-induced hepatotoxicity

In this study, kaempferol (KA) ameliorates APAP-induced hepatotoxicity by activating HO-1/NQO1 and inhibiting HMGB1/TLR4/NF-κB signaling and NLRP3 inflammasome activation.