Effect of glycyrrhizic acid and 18β-glycyrrhetinic acid on the differentiation of human umbilical cord-mesenchymal stem cells into hepatocytes

Abstract

BACKGROUND End-stage liver disease is a global health complication with high prevalence and limited treatment options. Cell-based therapies using mesenchymal stem cells (MSCs) emerged as an alternative approach to support hepatic regeneration. In vitro preconditioning strategies have been employed to strengthen the regenerative and differentiation potential of MSCs towards hepatic lineage. Chemical compounds of the triterpene class; glycyrrhizic acid (GA) and 18β-glycyrrhetinic acid (GT) possess diverse therapeutic properties including hepato-protection and anti-fibrosis characteristics. They are capable of modulating several signaling pathways that are crucial in hepatic regeneration. Preconditioning with hepato-protective triterpenes may stimulate MSC fate transition towards hepatocytes. AIM To explore the effect of GA and GT on hepatic differentiation of human umbilical cord-MSCs (hUC-MSCs). METHODS hUC-MSCs were isolated and characterized phenotypically by flow cytometry and immunocytochemistry for the expression of MSC-associated surface molecules. Isolated cells were treated with GA, GT, and their combination for 24 h and then analyzed at three time points; day 7, 14, and 21. qRT-PCR was performed for the expression of hepatic genes. Expression of hepatic proteins was analyzed by immunocytochemistry at day 21. Periodic acid Schiff staining was performed to determine the functional ability of treated cells. RESULTS The fusiform-shaped morphology of MSCs in the treatment groups in comparison with the untreated control, eventually progressed towards the polygonal morphology of hepatocytes with the passage of time. The temporal transcriptional profile of preconditioned MSCs displayed significant expression of hepatic genes with increasing time of differentiation. Preconditioned cells showed positive 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. CONCLUSION Preconditioning with glycyrrhizic acid, 18β-glycyrrhetinic acid and their combination, successfully differentiates hUC-MSCs into hepatic-like cells. These MSCs may serve as a better therapeutic option for degenerative liver diseases in future.

Ameliorative effect of licorice extract against the detrimental effect of glyphosate-based pesticide: Toxicity and health

“…At all-time points, the expression of the early hepatocyte marker AFP was found to be significantly upregulated in mesenchymal stem cells that were treated with GA or 18-glycyrrhetinic acid. The level of expression exhibited a progressive reduction over time [ 45 ].…”

3-Epi-betulinic acid 3- O- β- D-glucopyranoside (eBAG) induces autophagy by activation of AMP-activated protein kinase in hepatocellular carcinoma

“…1b). Triterpenoids reduce proliferation and promote death in multiple types of cancer cells [40] . To determine whether eBAG-regulated cell proliferation and death are generalized, we detected cell viability and cell death in human colon cancer HCT116 and esophageal cancer KYSE30 cells under the treatment of eBAG for 48 h. Similarly, exposure to eBAG markedly reduced the survival and enhanced the death of HCT116 and KYSE30 cells (Figs.…”

Mesenchymal stromal/stem cells and their extracellular vesicles in liver diseases: insights on their immunomodulatory roles and clinical applications

“…Furthermore, T-ADSCs showed better therapeutic effect on rats with liver injury than ADSCs due to significant suppression of pyroptosis as well as autophagy accompanied with intensive paracrine vascular endothelial growth factor from T-ADSCs [ 117 ]. In addition, glycyrrhizic acid and 18β-glycyrrhetinic acid possessed diverse therapeutic properties including hepatoprotective and anti-fibrotic characteristics, which facilitated the regenerative and differentiation potential of MSCs towards hepatocytes [ 118 ]. These results indicate that they could be developed as cytoprotective agents for successful MSCs transplantation in various liver diseases.…”