Gastroenterology

Gastroenterology

Volume 118, Issue 2, February 2000, Pages 380-389
Gastroenterology

Liver, Pancreas, and Biliary Tract
Genipin, a metabolite derived from the herbal medicine Inchin-ko-to, and suppression of Fas-induced lethal liver apoptosis in mice

https://doi.org/10.1016/S0016-5085(00)70220-4Get rights and content

Abstract

Background & Aims: We showed previously that a Kampo (Chinese/Japanese herbal) medicine, Inchin-ko-to (ICKT), inhibits hepatocyte apoptosis induced by transforming growth factor β1 in vitro. The present study investigated whether ICKT or its ingredients inhibit Fas-mediated liver apoptosis in vivo. Methods: Acute liver injury was induced by an intravenous injection of anti-Fas antibody, Jo2. The effects of ICKT and its ingredients on lethality, histology, apoptotic index, serum transaminase levels, caspase activation, mitochondrial membrane potential (ΔΨm), and mitochondrial permeability transition (MPT) were analyzed. Apoptosis in mouse hepatocytes in vitro was also evaluated. Results: Pretreatment with ICKT rescued 75% of Jo2-treated mice and markedly suppressed liver apoptosis/injury. Genipin, an intestinal bacterial metabolite of geniposide that is a major ingredient of ICKT, was found to be an active principle of ICKT. Genipin also suppressed in vitro Fas-mediated apoptosis in primary-cultured murine hepatocytes. Activation of caspase 3 and 8 in the liver homogenate and rapid reduction of ΔΨm of hepatocytes isolated from Jo2-treated mice were inhibited by genipin preadministration. The resistance to Ca2+-induced MPT was enhanced in liver mitochondria of genipin-treated mice. Conclusions: These results suggest that the antiapoptotic activity of genipin via the interference with MPT is a possible mechanism for therapeutic effects of ICKT.

GASTROENTEROLOGY 2000;118:380-389

Section snippets

Drugs, antibodies, and chemicals

Extract powders of ICKT and the Kampo medicine Sho-saiko-to were obtained from Tsumura & Co. (Tokyo, Japan). Glycyrrhizin (GLN) was purchased as Stronger Neo-Minophagen C from Minophagen Pharmaceutical Co. (Tokyo, Japan). Ursodeoxycholic acid was from Tokyo Tanabe Pharmaceutical Co. (Tokyo, Japan). A monoclonal anti-murine Fas antibody Jo2 and fluorescein isothiocyanate (FITC)-labeled Jo2 were obtained from Pharmingen (San Diego, CA). Other chemicals were purchased from Sigma Chemical Co. (St.

Protection from Fas-mediated death by ICKT

When 6.7 μg of the anti-Fas antibody Jo2 was injected IV in 4–5-week-old female BALB/c mice, severe hepatic failure rapidly developed and all control mice died within 24 hours. Two Kampo (Chinese/Japanese herbal) medicines, ICKT and Sho-saiko-to, and GLN and ursodeoxycholic acid, approved drugs for the treatment of liver diseases, were administered for 4 days before Jo2 injection. Fifteen of the 20 mice treated with 2 g/kg ICKT survived the antibody administration (Figure 1; P < 0.0001) and

Discussion

In this study we used a fulminant hepatitis model, first reported by Ogasawara et al.,3 that is based on massive liver cell apoptosis caused by IP or IV injection of the agonistic anti-Fas antibody Jo2. Using the same model, several researchers have examined the effects of apoptosis-modulating agents on Fas-mediated lethal liver injury. Mice overexpressing bcl-2, a gene with antiapoptotic properties, specifically in the liver, were protected from the lethality of Jo2 antibody treatment.27

References (46)

  • J Yamahara et al.

    Vascular dilatory action of Artemisia capillaris bud extracts and their active constituent

    J Ethnopharmacol

    (1989)
  • HL Li et al.

    Cleavage of bid by caspase 8 mediates the mitochondrial damage in the fas pathway of apoptosis

    Cell

    (1998)
  • X Luo et al.

    Bid, a bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors

    Cell

    (1998)
  • M Zoratti et al.

    The mitochondrial permeability transition

    Biochim Biophys Acta

    (1995)
  • I Marzo et al.

    Caspases disrupt mitochondrial membrane barrier function

    FEBS Lett

    (1998)
  • J Ogasawara et al.

    Lethal effect of the anti-Fas antibody in mice

    Nature

    (1993)
  • Y Nishimura et al.

    In vivo analysis of Fas antigen-mediated apoptosis: effects of agonistic anti-mouse Fas mAb on thymus, spleen and liver

    Int Immunol

    (1997)
  • H Tsutsui et al.

    IL-18 accounts for both TNF-alpha– and Fas ligand–mediated hepatotoxic pathways in endotoxin-induced liver injury in mice

    J Immunol

    (1997)
  • R Ksontini et al.

    Disparate roles for TNF-alpha and Fas ligand in concanavalin A–induced hepatitis

    J Immunol

    (1998)
  • T Kondo et al.

    Essential roles of the Fas ligand in the development of hepatitis

    Nat Med

    (1997)
  • P Bobe et al.

    Fas-mediated liver damage in MRL hemopoietic chimeras undergoing Ipr-mediated graft-versus-host disease

    J Immunol

    (1997)
  • M Adachi et al.

    Targeted mutation in the Fas gene causes hyperplasia in peripheral lymphoid organs and liver

    Nat Genet

    (1995)
  • H Yano et al.

    The herbal medicine Sho-saiko-to inhibits proliferation of cancer cell lines by inducing apoptosis and arrest at the G0/G1 phase

    Cancer Res

    (1994)
  • Cited by (0)

    Address requests for reprints to: Masahiro Yamamoto, Ph.D., Kampo Pharmacology Department, Tsumura Central Research Laboratories, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki-gun, Ibaraki 300-1192, Japan. e-mail: [email protected]; fax: (81) 298-89-3867.

    View full text