AT-MSCs Antifibrotic Activity is Improved by Eugenol through Modulation of TGF-β/Smad Signaling Pathway in Rats.

Moustafa Fathy,Toshiko Yoshida,Heba M Saad Eldien,Motonori Okabe

doi:10.3390/molecules25020348

Abstract

For hepatic failure, stem cell transplantation has been chosen as an alternative therapy, especially for mesenchymal stem cells (MSCs). The aim of this study was to investigate the effect of eugenol (EUG) on the in vivo antifibrotic activity of adipose tissue-derived MSCs (AT-MSCs) and the underlying mechanism. After characterization of MSCs, rats were divided into five groups, Group 1 (normal control), Group 2 (CCl4), Group 3 (CCl4 + AT-MSCs), Group 4 (CCl4 + EUG) and Group 5 (CCl4 + AT-MSCs + EUG). Biochemical and histopathological investigations were performed. Furthermore, expression of type 1 collagen, α-SMA, TGF-β1, Smad3 and P-Smad3 was estimated. Compared to the single treatment with AT-MSCs, the combination treatment of the fibrotic rats with AT-MSCs and EUG significantly improved the plasma fibrinogen concentration, IL-10 level and proliferating cell nuclear antigen expression, and also significantly decreased the serum levels of liver enzymes, IL-6, IL-1β, TNF-α, type III collagen, hyaluronic acid, hydroxyproline and the TGF-β growth factor. Furthermore, the combination treatment significantly decreased the hepatic expression of fibrotic markers genes (Type 1 collagen and α-SMA) and proteins (α-SMA, TGF-β1 and phospho-Smad3) more than the treatment with AT-MSCs alone. We demonstrated that the combination treatment with EUG and AT-MSCs strongly inhibited the advancement of CCl4-induced hepatic fibrosis, compared with AT-MSCs alone, through TGF-β/Smad pathway inhibition. This approach is completely novel, so more investigations are necessary to improve our perception of the underlying molecular mechanisms accountable for the effects of EUG on the antifibrotic potential of AT-MSCs.

Highlights

For hepatic failure, stem cell transplantation has been chosen as an alternative therapy.Mesenchymal stem cells (MSCs), derived from different organs, have much superiority in regenerative medicine, including multipotent differentiation capacity, their low immunogenic properties and high proliferation rate
Cell surface markers expression of Adipose tissue (AT)-mesenchymal stem cells (MSCs) was examined by flow cytometric (FCM) analysis for MSC markers (CD90, CD105 and CD73) and hematopoietic markers (CD34 and CD45)
Downregulation of the fibrotic genes (Type 1 collagen and α-Smooth Muscle Actin (α-SMA)) and the growth factor (TGF-β1) expression, which was evaluated by quantitative RT-PCR, was significantly associated with the combination treatment more than that with the single treatment with adipose tissue-derived MSCs (AT-MSCs), indicating decreased hepatic fibrosis

Summary

Introduction

Mesenchymal stem cells (MSCs), derived from different organs, have much superiority in regenerative medicine, including multipotent differentiation capacity, their low immunogenic properties and high proliferation rate. They have the ability to drift toward the microenvironments of injured sites [1,2,3]. MSCs present many other features, like no adverse reactions related to MSCs transplant, and long-term storage without considerable loss of potency [4] They are immunomodulatory, can directly differentiate to new hepatocytes, have antifibrotic properties, can promote tissue repair and inhibit hepatic stellate cells activation [5].

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