Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Oct 26, 2021; 13(10): 1580-1594
Published online Oct 26, 2021. doi: 10.4252/wjsc.v13.i10.1580
Effect of glycyrrhizic acid and 18β-glycyrrhetinic acid on the differentiation of human umbilical cord-mesenchymal stem cells into hepatocytes
Abiha Fatima, Tuba Shakil Malick, Irfan Khan, Aisha Ishaque, Asmat Salim
Abiha Fatima, Tuba Shakil Malick, Irfan Khan, Aisha Ishaque, Asmat Salim, Stem Cell Laboratory, Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi 75270, Sindh, Pakistan
Author contributions: Fatima A performed all experiments, analyzed the data and wrote the first draft; Malick TS and Ishaque A performed part of the experiments and analysis of the data; Khan I provided technical guidance for all experiments and data analysis; Salim A designed the research study, supervised the research and wrote the manuscript in its final form; all authors have read and approved the final manuscript.
Institutional review board statement: Present study was approved by the Independent Ethics Committee, International Center for Chemical and Biological Sciences (IEC, ICCBS) under protocol #: ICCBS/IEC-037-HT-2018/Protocol/1.0.
Conflict-of-interest statement: The authors confirm that this article has no conflict of interest to report.
Data sharing statement: No additional data is available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Asmat Salim, PhD, Professor, Stem Cell Laboratory, Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi 75270, Sindh, Pakistan. asmat.salim@iccs.edu
Received: April 13, 2021
Peer-review started: April 13, 2021
First decision: May 12, 2021
Revised: May 25, 2021
Accepted: September 19, 2021
Article in press: September 19, 2021
Published online: October 26, 2021
Processing time: 195 Days and 17.4 Hours
ARTICLE HIGHLIGHTS
Research background

End-stage liver disease (ESLD) causes extensive healthcare burden with limited treatment options. Cell-based therapies using mesenchymal stem cells (MSCs) emerged as a potential approach to support hepatic regeneration in ESLD. However, effective translation of this therapy requires MSCs to have maximum regenerative potential. In vitro preconditioning strategies have been employed to strengthen the regenerative and differentiation potential of MSCs. Chemical compounds of the class triterpenes; glycyrrhizic acid and 18β-glycyrrhetinic acid present diverse therapeutic features including hepato-protection and anti-fibrotic characteristics. They are capable of modulating several physiological pathways that are crucial in hepatic regeneration. Preconditioning with hepato-protective triterpenes may stimulate MSC fate transition towards hepatocytes.

Research motivation

Although mesenchymal stem cell-mediated therapy has proved as an effective approach for hepatic regeneration, the major challenge is low cell viability and resistance in the impaired tissue post-transplantation. To acquire a persistent therapeutic efficacy, in vitro preconditioning of MSCs with hepato-protective chemical compounds may facilitate their regenerative and differentiation potential towards hepatic lineage for better survival, homing, and migration ability at the site of injury.

Research objectives

Considering the characteristics of triterpenes, glycyrrhizic acid and 18β-glycyrrhetinic acid in hepatic anomalies, the objective of the study is to explore their role in hepatic differentiation of MSCs. Preconditioned cells may serve as a better source for tissue regeneration in liver injury.

Research methods

hUC-MSCs were harvested and characterized phenotypically by flow cytometry and immunocytochemistry for the expression of MSC associated surface molecules. Isolated cells were treated with glycyrrhizic acid, 18β-glycyrrhetinic acid, and their combination for 24 h, and then analyzed at three time points; day 7, 14, and 21. qRT-PCR was performed to evaluate the expression of hepatic genes. On day 21, hepatic proteins were analyzed by immunocytochemistry. Periodic acid Schiff staining was performed to determine the functional ability of treated cells.

Research results

The transcriptional profile of preconditioned MSCs displayed significant expression of hepatic genes with increasing time of differentiation. Preconditioned cells showed positive protein expression of hepatocyte specific proteins. The results were further corroborated by positive periodic acid Schiff staining, indicating the presence of glycogen in their cytoplasm. Moreover, bi-nucleated cells which is the typical feature of hepatocytes, were also seen in the preconditioned cells.

Research conclusions

Our data suggest that preconditioning of hUC-MSCs with triterpene compounds, glycyrrhizic acid, and 18β-glycyrrhetinic acid or both, successfully differentiates these cells into hepatic-like cells. The study would serve as an attempt to develop new targeted therapies using triterpenes in combination with stem cells for the treatment of end-stage liver diseases.

Research perspectives

The present study is an endeavor to augment cell based therapeutic approach by preconditioning hUC-MSCs with glycyrrhizic acid and 18β-glycyrrhetinic acid to promote their therapeutic and differentiation potential towards hepatic lineage. The preconditioned MSCs may serve as an effective source for cell therapy for injured hepatic tissue in clinical applications.