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Apoptosis

, Volume 20, Issue 10, pp 1296–1306 | Cite as

Morin mitigates acetaminophen-induced liver injury by potentiating Nrf2 regulated survival mechanism through molecular intervention in PHLPP2-Akt-Gsk3β axis

  • Fatima Rizvi
  • Alpana Mathur
  • Poonam KakkarEmail author
Original Paper

Abstract

Acetaminophen (APAP) is frequently taken to relieve pain. Staggered APAP overdoses have been reported to cause acetaminophen-induced liver injury (AILI). Identification of efficacious therapeutic modalities to address complications imposed by accidental/intentional long-term APAP ingestion is needed. Morin, a plant-derived phytochemical, possesses a multitude of pharmacological properties including hepatoprotective action; however, the underlying mechanisms have been inadequately explored. Our present report demonstrates significant attenuation of APAP-mediated liver injury by morin supplementation in vivo as indicated by reduction in histological and serum markers of hepatotoxicity. Morin not only limited necroinflammation as revealed by reduced HMGB1 release, NALP3 and caspase-1 maturation, but also suppressed oxidative stress and mitochondrial dysfunction. This suggests that morin may have exerted its cytoprotective role by way of early intervention in the pathway leading to perpetuation of AILI. Morin reinforced cellular defenses by suppressing Nrf2 ubiquitination and promoting nuclear Nrf2 retention as well as ARE-Nrf2 binding affinity. The effects were observed to be a result of molecular intervention in the activity of PHLPP2, a phosphatase previously reported by us to subdue cellular Nrf2 responses via Fyn kinase activation. Morin was observed to inhibit APAP-induced increase in PHLPP2 activity ex vivo as well as its association with cellular target Akt1. As a result, morin prevented oxidative stress induced deactivation of Akt (Ser473) leading to suppression in GSK3β and Fyn kinase activation. The study supports the inhibitory action of morin against PHLPP2-regulated Nrf2-suppression and hence indentifies Nrf2-potentiating property of morin that may be exploited in developing novel therapeutic strategy to address AILI.

Keywords

PHLPP2 Nrf2 Acetaminophen Fyn kinase Oxidative stress Flavonoid 

Abbreviations

AIF

Apoptosis inducing factor

AILI

Acetaminophen-induced liver injury

ALP

Alkaline phosphatase

ALT

Alanine aminotransferase

APAP

Acetaminophen

ARE

Antioxidant redox element

AST

Aspartate transaminase

ATP

Adenosine triphosphate

CAT

Catalase

CMC

Carboxymethylcellulose

DAMPs

Damage-associated molecular pattern molecules

DCFH-DA

2′,7′-Dichlorofluorescein diacetate

DHE

Dihydroethidium

ELISA

Enzyme-linked immunosorbent assay

EndoG

Endonuclease G

FITC

Fluorescein isothiocyanate

GAPDH

Glyceraldehyde phosphate dehydrogenase

GPx

Glutathione peroxidase

GR

Glutathione reductase

GSH

Glutathione

GSSG

Glutathione disulfide

GST

Glutathione S-transferase

H&E

Hematoxylin and eosin

HMGB1

High-mobility group box 1

HO1

Hemeoxygenase 1

JC-1

5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolyl carbocyanine iodide

Keap1

Kelch-like ECH-associated protein-1

LDH

Lactate dehydrogenase

MTT

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NAC

N-Acetylcysteine

NAD

Nicotinamide adenine dinucleotide

NADH

Nicotinamide adenine dinucleotide reduced

NALP3

NACHT, LRR and PYD domains-containing protein 3

NAPQI

N-Acetyl-p-benzoquinoneimine

NQO1

NAD(P)H quinine oxidoreductase 1

Nrf2

Nuclear factor erythroid 2-related factor 2

NSAIDs

Non-steroidal anti-inflammatory drugs

PHLPP2

PH-domain and Leucine rich repeat protein phosphatase 2

PI

Propidium iodide

PVDF

Polyvinylidene fluoride

RBC

Red blood cell

ROS

Reactive oxygen species

SEM

Standard error of the mean

SOD

Superoxide dismutase

TrxRed

Thioredoxin reductase

Notes

Acknowledgments

We are grateful to the Institutional Manuscript Review Committee of CSIR-IITR for allotting communication number 3253 for the manuscript. Financial support was received from CSIR Network Project-BSC 0111 and BSC 0106. First author acknowledges Council of Scientific and Industrial Research (CSIR), India, for award of fellowship.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

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Supplementary material 1 (TIFF 1110 kb)
10495_2015_1160_MOESM2_ESM.tif (318 kb)
Supplementary material 2 (TIFF 318 kb)
10495_2015_1160_MOESM3_ESM.docx (29 kb)
Supplementary material 3 (DOCX 29 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Academy of Scientific and Innovative ResearchLucknowIndia
  2. 2.Herbal Research Section, Food Drug and Chemical Toxicology DivisionCouncil of Scientific and Industrial Research-Indian Institute of Toxicology Research (CSIR-IITR)LucknowIndia

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