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Modulation of mercury-induced mitochondria-dependent apoptosis by glycine in hepatocytes

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Abstract

Mercury (Hg) is one of the universal environmental pollutants and is responsible for various organ pathophysiology including oxidative stress-induced hepatic disorders. In the present study, we aimed to explore the protective role of glycine in Hg-induced cytotoxicity and cell death in murine hepatocytes. Exposure of mercury (20 μM), in the form HgCl2 for 1 h, significantly enhanced the ALT and ALP leakage, increased reactive oxygen species production, reduced cell viability and distorted the antioxidant status of hepatocytes. Flow cytometric analyses shows that Hg-induced apoptotic death in hepatocytes. Mechanism of this pathophysiology involves reduced mitochondrial membrane potential, variations in Bcl-2/Bad proteins, activation of caspases and cleavage of PARP protein. In addition, Hg distinctly increased NF-κB phosphorylation in association with IKKα phosphorylation and IκBα degradation. Concurrent treatment with glycine (45 mM), however, reduced Hg-induced oxidative stress, attenuated the changes in NF-κB phosphorylation and protects hepatocytes from Hg-induced apoptotic death. Hg also distinctly increased the phosphorylation of p38, JNK and ERK mitogen-activated protein kinase (MAPKs). Glycine treatment suppressed these apoptotic events, signifying its protective role in Hg-induced hepatocyte apoptosis as referred by reduction of p38, JNK and ERK MAPK signaling pathways. Results suggest that glycine can modulate Hg-induced oxidative stress and apoptosis in hepatocytes probably because of its antioxidant activity and functioning via mitochondria-dependent pathways and could be a beneficial agent in oxidative stress-mediated liver diseases.

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Abbreviations

ALP:

Alkaline phosphatase

ALT:

Alanine aminotranferase

CAT:

Catalase

DMEM:

Dulbecco’s modified eagle’s medium

DMSO:

Dimethyl sulphoxide

GSH:

Glutathione

GSSG:

Glutathione disulfide

GPx:

Glutathione peroxidase

GST:

Glutathione S-transferase

GR:

Glutathione reductase

HgCl2 :

Mercuric chloride

NF-κB:

Nuclear factor kappa B

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

PARP:

Poly ADP ribose polymerase

MAPK:

Mitogen-activated protein kinase

ERK:

Extracellular-signal-regulated kinase

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Acknowledgments

The authors are grateful to Mr. Prasanta Pal for excellent technical assistance for the study.

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The authors have declared that no conflict of interest exists.

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Authors and Affiliations

  1. Division of Molecular Medicine, Bose Institute, P-1/12, CIT Scheme VII M, Kolkata, 700054, West Bengal, India

    Pabitra Bikash Pal, Sankhadeep Pal, Joydeep Das & Parames C. Sil

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  1. Pabitra Bikash Pal
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  2. Sankhadeep Pal
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  3. Joydeep Das
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  4. Parames C. Sil
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Correspondence to Parames C. Sil.

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Pal, P.B., Pal, S., Das, J. et al. Modulation of mercury-induced mitochondria-dependent apoptosis by glycine in hepatocytes. Amino Acids 42, 1669–1683 (2012). https://doi.org/10.1007/s00726-011-0869-3

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