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Proline protects liver from d-galactosamine hepatitis by activating the IL-6/STAT3 survival signaling pathway

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Abstract

The oral administration of proline, one of the non-essential amino acids, has been shown to effectively protect the liver from d-galactosamine (GalN)-induced liver injury and to improve the survival rate. The aim of this study was to investigate the mechanism of this protective action of proline. We paid particular attention to the effect of proline on inflammatory activation, regenerative response, and the associated signal transduction in the liver. Male Fischer rats received intraperitoneal injections of GalN (1.4 g/kg) with or without the oral administration of proline (2 g/kg) 1 h before GalN treatment. Liver pathology, plasma indices of inflammation, and the level of proliferative marker in the liver were monitored. The hepatic activation of interleukin-6 (IL-6)/signal transducer and activator of transcription (STAT)-3 pathway, which is downstream of tumor necrosis factor (TNF)-α/nuclear factor-κB, was also studied. GalN induced massive inflammatory expansion in the liver, leading to a high death rate (60 %) more than 72 h after the treatment. Proline administration significantly suppressed inflammatory infiltration in the live after 48 h, which was accompanied by depletion of plasma TNF-α, glutamic oxaloacetic transaminase, and glutamic pyruvic transaminase. The mRNA expression of histone H3, a marker of proliferation, was significantly upregulated in the liver of proline-treated animals. Furthermore, IL-6/STAT-3 pathway, an anti-inflammatory and regenerative signaling pathway, was strongly activated prior to these observations, with the upregulated expression of downstream genes. These results suggest that the tissue-protective mechanism of proline involves the early activation of IL-6/STAT-3 pathway in the liver, with subsequent activation of the regenerative response and suppression of massive inflammatory activation.

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Abbreviations

GalN:

d-Galactosamine

IL-6:

Interleukin-6

STAT3:

Signal transducer and activator of transcription 3

TNF-α:

Tumor necrosis factor-α

NF-κB:

Nuclear factor kappa B

GOT:

Glutamic oxaloacetic transaminase

GPT:

Glutamic pyruvic transaminase

LEC rat:

Long Evans Cinnamon rat

SDS:

Sodium dodecyl sulfate

XIAP:

X-chromosome-linked inhibitor of apoptosis

MAPK:

Mitogen-activated protein kinase kinase

FLIP:

FLICE-inhibitory protein

ROS:

Reactive oxygen species

POX:

Proline oxidase

FAD:

Flavine adenine dinucleotide

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Acknowledgments

This study was supported by a private fund from Ajinomoto Co., Inc. The authors thank Dr. Koichi Ishii, Mr. Katsunori Kobayashi, Dr. Ikuko Sasahara, D.V.M., and Ms. Miwako Yamanoue for technical assistance and significant support.

Conflict of interest

This study was supported by a private fund from Ajinomoto Co., Inc. All authors are employee of Ajinomoto Co., Inc., which has published a large number of papers about the function of amino acids, strenuously doing basic research. No patent has been applied based on the research of this manuscript.

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  1. Yoko Obayashi

    Present address: , 1-15-1, Kyobashi, Chuo-ku, Tokyo, 104-8315, Japan

Authors and Affiliations

  1. Research Institute for Health Fundamentals, Ajinomoto Co., Inc, 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, Kanagawa, 210-8681, Japan

    Yoko Obayashi, Harumi Arisaka, Shintaro Yoshida, Masato Mori & Michio Takahashi

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  1. Yoko Obayashi
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Correspondence to Yoko Obayashi.

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Obayashi, Y., Arisaka, H., Yoshida, S. et al. Proline protects liver from d-galactosamine hepatitis by activating the IL-6/STAT3 survival signaling pathway. Amino Acids 43, 2371–2380 (2012). https://doi.org/10.1007/s00726-012-1317-8

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