Diammonium Glycyrrhizinate Mitigates Liver Injury Via Inhibiting Proliferation Of NKT Cells And Promoting Proliferation Of Tregs

Drug Des Devel Ther. 2019 Oct 16:13:3579-3589. doi: 10.2147/DDDT.S220030. eCollection 2019.

Abstract

Purpose: Diammonium glycyrrhizinate (DG) is a replacement for glycyrrhizic acid, which is used as a hepatic protector in clinical practice for most liver diseases. The potential role of immune response during autoimmune hepatitis-induced by concanavalin A (Con A)-remains to be elucidated.

Methods: C57BL/6J mice were treated with two different doses of DG (75 and 200 mg/kg) 2 hrs before administering Con A. The mice were sacrificed after administering Con A for 0, 6, and 24 hrs. Liver damage grade and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin levels were evaluated. The expression level of cleaved-caspase 3 in liver was detected by Western blotting. Inflammatory cytokines such as tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and interferon γ (IFN-γ) in liver were detected by RT-PCR. Thymus, peripheral blood, spleen, and liver tissues were collected to analyze the percentages of NKT cells, subsets of CD4+CD25-CD69+ and CD8+CD69+ T cells, and subsets of regulatory T cells (Tregs).

Results: Our results revealed that DG pre-treatment significantly decreased the serum ALT and AST levels and improved the histological damage in Con A-induced autoimmune liver injury. Pre-treatment with DG down-regulated the inflammatory cytokines upon challenge with Con A. The DG pre-treatment inhibited the apoptosis of T lymphocytes in the thymus. Further, it effectively suppressed the proliferation of CD4+CD25-CD69+ and CD8+CD69+ subsets in the peripheral blood and spleen. In addition, the DG pretreatment significantly downregulated the frequency of NKT cells, while upregulating the frequency of Tregs in the liver.

Conclusion: We believe that the potential protective effect of DG against Con A-induced hepatitis may be partially attributed to its inhibitory activities on inflammatory cytokines in the livers, lymphocyte apoptosis in the thymus, NKT cells proliferation, and activation of CD8+T cells; further, there may also be a possibility of DC promoting Tregs proliferation.

Keywords: NKT cells; autoimmune hepatitis; concanavalin A; diammonium glycyrrhizinate; regulatory T-cell.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Cell Proliferation / drug effects
  • Chemical and Drug Induced Liver Injury / drug therapy*
  • Chemical and Drug Induced Liver Injury / pathology
  • Concanavalin A
  • Disease Models, Animal
  • Dose-Response Relationship, Drug
  • Glycyrrhizic Acid / pharmacology*
  • Hepatitis, Autoimmune / drug therapy*
  • Hepatitis, Autoimmune / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Structure
  • Natural Killer T-Cells / drug effects*
  • Natural Killer T-Cells / pathology
  • Structure-Activity Relationship
  • T-Lymphocytes, Regulatory / drug effects*
  • T-Lymphocytes, Regulatory / pathology

Substances

  • Anti-Inflammatory Agents
  • Concanavalin A
  • Glycyrrhizic Acid