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Reverse pharmacology of Nimbin-N2 attenuates alcoholic liver injury and promotes the hepatoprotective dual role of improving lipid metabolism and downregulating the levels of inflammatory cytokines in zebrafish larval model

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Abstract

Alcoholic liver disease is one of the most prominent liver diseases in the world. Lipid accumulation accompanied by oxidative stress and inflammation in the liver is the most important pathogenesis of ALD. This study was designed to investigate the anti-oxidative, fat metabolism-regulating, and anti-inflammatory potential of N2, a seminatural analog of Nimbin. The ethanol exposure was found to induce liver injury on zebrafish larvae, such as liver inflammation, lipid accumulation, oxidative stress, and hepatocytes apoptosis. N2 was subjected to ADMET screening in-silico, and it was observed N2’s co-exposure decreased the ROS, apoptosis, lipid peroxidation, and macrophage accumulation in the liver of larval zebrafish. To further study the mechanism behind ethanol hepatotoxicity and the hepatoprotective behavior of N2, gene expression changes were determined in zebrafish. The results of this study revealed that ethanol exposure upregulated mRNA expressions of SREBP1, C/EBP-α, FAS and provoked more severe oxidative stress and hepatitis via upregulation of inflammatory cytokines TNF-α, IL-10, IL-1β, iNOS, COX-2. However, the N2 co-exposure protected the hepatocyte damage and almost reversed the condition by downregulating the mRNA levels. The study suggested that N2 could be an effective therapeutic agent for the treatment of ALD and other inflammatory conditions.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Code availability

Not applicable.

Abbreviations

ALD:

Alcoholic liver disease

ADMET:

Adsorption, distribution, metabolism, excretion, toxicity.

SMILES:

Simplified molecular-input line-entry system

DCF-DA:

Dichlorodihydrofluorescein diacetate

ROS:

Reactive oxygen species

DNA:

Deoxyribonucleic acid

RNA:

Ribonucleic acid

DPPP:

Diphenyl-1-pyrenylphosphine

SOD:

Superoxide dismutase

CAT:

Catalase

PBS:

Phosphate-buffered saline

CMC-Na:

Sodium carboxymethyl cellulose salt

HBD:

Hydrogen-bond donors

HBA:

Hydrogen-bond acceptor

TPSA:

Total polar surface area

GI:

Gastrointestinal

BBB:

Blood–brain barrier

CYP:

Cytochrome

P-gp:

P-glycoprotein

TC:

Total cholesterol

TG:

Total triglyceride

SREBP1:

Sterol regulatory element-binding transcription factor 1

C/EBP- α:

Enhancer Binding Protein Alpha

FAS:

Fatty acid synthase

iNOS:

Induced Nitric oxide synthases

COX-2:

Cyclooxygenase-2

TNF-α:

Tumor necrosis factor-alpha

IL-10:

Interleukin

AFLD:

Alcoholic fatty liver disease

GAS:

Gastrodin

NASH:

Non-alcoholic steatohepatitis

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Acknowledgements

Dr. Pushparathinam Gopinath thank and gratefully acknowledge the Science and Engineering Research Board (SERB), Government of India for the financial support under the start-up research grant (SERB-SRG/2019/001133). He also thanks the Department of Chemistry, SRMIST for providing the necessary infrastructure facility to complete the study. The authors express their sincere appreciation to the Researchers Supporting Project Number (RSP2022R414) King Saud University, Riyadh, Saudi Arabia.

Funding

This study was supported by Science and Engineering Research Board (SERB), Government of India through start-up research grant (No. SERB-SRG/2019/001133) and Researchers Supporting Project (Number: RSP2022R414), King Saud University, Riyadh, Saudi Arabia.

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Authors and Affiliations

  1. Department of Biotechnology, College of Science and Humanities, SRM Institute of Science and Technology, Kattankulathur 603 203, Chennai, Tamil Nadu, India

    Gokul Sudhakaran, Ajay Guru, B. Haridevamuthu, Raghul Murugan & Jesu Arockiaraj

  2. Department of Chemistry, College of Engineering and Technology, SRM Institute of Science and Technology, Kattankulathur 603 203, Chennai, Tamil Nadu, India

    Pandurangan Prathap & Pushparathinam Gopinath

  3. Department of Zoology, College of Science, King Saud University, PO Box 2455, Riyadh, 11451, Saudi Arabia

    Bader O. Almutairi & Mikhlid H. Almutairi

  4. Institute for Cellular and Molecular Biology, The University of Texas at Austin, University Station A4800, Austin, TX, 78712, USA

    Annie Juliet

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  1. Gokul Sudhakaran
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Contributions

Conceived and designed the experiments, All Authors; Performed the experiments, GS, PP, AG: Analyzed the data, All Authors; Contributed reagents/materials/analysis tools and Supervised the research, BOA, MHA, AJ, PG, JA. Wrote the paper, All Authors.

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Correspondence to Jesu Arockiaraj.

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Sudhakaran, G., Prathap, P., Guru, A. et al. Reverse pharmacology of Nimbin-N2 attenuates alcoholic liver injury and promotes the hepatoprotective dual role of improving lipid metabolism and downregulating the levels of inflammatory cytokines in zebrafish larval model. Mol Cell Biochem 477, 2387–2401 (2022). https://doi.org/10.1007/s11010-022-04448-7

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