Hepatoprotective Activity of InlB321/15, the HGFR Ligand of Bacterial Origin, in CCI4-Induced Acute Liver Injury Mice.

Chalenko Chalenko,Sysolyatina Sysolyatina,Lavrikova Lavrikova,Midiber Midiber,Mikhaleva Mikhaleva,Ermolaeva Ermolaeva,Kalinin Kalinin,Sobyanin Sobyanin

doi:10.3390/biomedicines7020029

Abstract

HGF (hepatocyte growth factor)/HGFR (HGF receptor) signaling pathway is a key pathway in liver protection and regeneration after acute toxic damage. Listeria monocytogenes toxin InlB contains a HGFR-interacting domain and is a functional analog of HGF. The aim of this work was to evaluate the hepatoprotective activity of the InlB HGFR-interacting domain. The recombinant HGFR-interacting domain InlB321/15 was purified from E. coli. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test was used to measure InlB321/15 mitogenic activity in HepG2 cells. Activation of MAPK- and PI3K/Akt-pathways was tracked with fluorescent microscopy, Western blotting, and ELISA. To evaluate hepatoprotective activity, InlB321/15 and recombinant human HGF (rhHGF) were intravenously injected at the same concentration of 2 ng·g−1 to BALB/c mice 2 h before liver injury with CCl4. InlB321/15 caused dose-dependent activation of MAPK- and PI3K/Akt-pathways and correspondent mitogenic effects. Both InlB321/15 and rhHGF improved macroscopic liver parameters (liver mass was 1.51, 1.27 and 1.15 g for the vehicle, InlB321/15 and rhHGF, respectively, p < 0.05), reduced necrosis (24.0%, 16.18% and 21.66% of the total area for the vehicle, InlB321/15 and rhHGF, respectively, p < 0.05). Obtained data suggest that InlB321/15 is a promising candidate for a tissue repair agent.

Highlights

Proliferation of hepatocytes and liver stem cells underlies liver regeneration after acute toxic damage [1]
InlB321/15 or recombinant human HGF (rhHGF) for 15 min, labelled with Hoechst to label nuclei, phospho-extracellularsignal-regulated kinase 1/2 (Erk1/2)-specific antibodies and Alexa Fluor 488-labelled secondary antibodies, and observed with fluorescent microscopy; scale bar 50 μm; (C) Coomassie R-250 stained SDS-PAGE and Western blotting with anti-phospho-Erk1/2, anti-phospho-Akt and anti-tubulin specific antibodies, cell lysates were obtained from cells treated with
We demonstrated that the InlB321/15 protein, which is a HGFR ligand derived from the clinical L. monocytogenes strain, is a potential inducer of both MAPK and PI3K/Akt pathways

Summary

Introduction

Proliferation of hepatocytes and liver stem cells underlies liver regeneration after acute toxic damage [1]. The HGF (hepatocyte growth factor)/HGFR (HGF receptor) signaling pathway is central to mitogenic, motogenic, and morphogenic effects in hepatocytes and hepatic progenitor cells [2,3]. Loss of the tyrosine kinase HGFR results in delay and abnormalities in the liver regeneration process [4]. Overexpression of HGF decreases acute liver injury and improves regeneration via activation of MAPK and PI3K/Akt/mTor signaling pathways and the antioxidant response [5,6]. The biological actions that are driven by the HGF/HGFR pathway all play roles in the progression of invasive and metastatic cancers [7,8]. Some pathogenic microorganisms hijack the host HGF/HGFR system to establish a comfortable environment for infection [8,9]

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