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Beneficial effects of combined ursodeoxycholic acid and angiotensin-II type 1 receptor blocker on hepatic fibrogenesis in a rat model of nonalcoholic steatohepatitis

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
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Abstract

Background and Aims

Ursodeoxycholic acid (UDCA) is considered to be effective in the treatment of nonalcoholic steatohepatitis (NASH), particularly in combination with other pharmacological agents. UDCA reportedly counteracts the effects of endotoxemia. Previously, we demonstrated attenuated hepatic fibrogenesis and suppression of activated hepatic stellate cells (Ac-HSC) with an angiotensin-II (AT-II) type 1 receptor blocker (ARB). Here we evaluated the simultaneous effect of both agents on hepatic fibrogenesis in a rat model of NASH.

Methods

Fischer 344 rats were fed a choline-deficient l-amino-acid-defined (CDAA) diet for 8 weeks. The therapeutic effect of UDCA and ARB was evaluated along with hepatic fibrogenesis, lipopolysaccharide (LPS)-Toll-like receptor 4 (TLR4) regulatory cascade, and intestinal barrier function. The direct inhibitory effect of both UDCA and ARB on Ac-HSC was assessed in vitro.

Results

Both UDCA and ARB had a potent inhibitory effect on hepatic fibrogenesis with suppression of the HSC activation and hepatic expression of transforming growth factor (TGF)-β1 and TLR4. UDCA decreased intestinal permeability by ameliorating zonula occuludens-1 (ZO-1) disruption induced by the CDAA diet. ARB was found to directly suppress regulation of Ac-HSC.

Conclusions

UDCA and ARB have a synergistic repressive effect on hepatic fibrogenesis by counteracting endotoxemia induced by intestinal barrier dysfunction and suppressing the activation of Ac-HSC. Because both agents are currently used in clinical practice, combined UDCA and ARB may represent a promising novel therapeutic approach for NASH.

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Authors and Affiliations

  1. Third Department of Internal Medicine, Nara Medical University, Shijo-cho 840, Kashihara, Nara, 634-8522, Japan

    Tadashi Namisaki, Ryuichi Noguchi, Kei Moriya, Mitsuteru Kitade, Yosuke Aihara, Akitoshi Douhara, Norihisa Nishimura, Kosuke Takeda, Yasushi Okura, Hideto Kawaratani, Hiroaki Takaya, Kenichiro Seki & Hitoshi Yoshiji

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  1. Tadashi Namisaki
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  2. Ryuichi Noguchi
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Correspondence to Tadashi Namisaki.

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535_2015_1104_MOESM1_ESM.pptx

Fig. S1 Inhibitory effect of UDCA and ARB on hepatic expression of connective tissue growth factor and alpha-2-HS-glycoprotein during hepatic fibrosis. (A) Hepatic expression of CTGF was elevated in the CDAA fed rats (G2) compared to CSAA fed rats (G1).The CDAA‑induced increase was significantly inhibited by treatment with UDCA (G3) and ARB (G4) compared with CDAA control group. Similar to the effect on hepatic fibrosis, the combination treatment with both agents exerted a more potent inhibitory effect on CTGF expression than that of either agent alone, and these inhibitory effects were almost parallel to the fibrosis area reduction. (B) Hepatic mRNA expression of alpha-2-HS-glycoprotein (Fetuin A/AHSG), surrogate marker for hepatic steatosis significantly increased in G2 and the CDAA-induced increase in AHSG significantly reduced in UDCA-treated rats (G3) and UDCA- and ARB-treated rats (G5). However, no significant reduction was found in ARB-treated rats (G4). Values are mean±SD (bars; n = 10). Asterisks indicate statistically significant difference between indicated experimental groups (* p < 0.05, ** p < 0.01). G1, CSAA-treated control group; G2, CDAA with vehicle-treated group; G3, CDAA with UDCA-treated group; G4, CDAA with ARB-treated group; G5, CDAA with UDCA- and ARB-treated group. (PPTX 64 kb)

535_2015_1104_MOESM2_ESM.pptx

Fig. S2 Inhibitory effect of UDCA and ARB on the TLR4 protein expression in the liver and the intestine. (A) The hepatic TLR4 protein expression increased in the CDAA diet group (G2). This increase was significantly suppressed by treatment with UDCA and ARB.The combination treatment of both agents (G4) exerted a more potent inhibitory effect than that of either agent alone. These inhibitory effects were almost parallel to the fibrosis area reduction. (B) However, no significant differences were detected in protein expression of intestinal TLR4 between G1 and G2. Values are represented as mean ± SD (bars; n = 10). Asterisks indicate statistically significant difference between indicated experimental groups (* p < 0.05, ** p < 0.01) (PPTX 386 kb)

535_2015_1104_MOESM3_ESM.pptx

Fig. S3 The inhibitory effect of UDCA and ARB on the expression of hepatic inflammatory cytokines, tumor necrosis factor-alpha (A), Interferon-gamma (B) and Interleukin-6 (C) using ELISA. All three hepatic proinflammatory cytokines were augmented in CDAA fed group. These diet-induced alterations in these pro-inflammatory cytokines were suppressed in UDCA treated group and combination treatment group (G5), whereas treatment with ARB did not significantly alter the expression of the three proinflammatory cytokines. Values are represented as mean ± SD (bars; n = 10). Asterisks indicate statistically significant difference between indicated experimental groups (** p < 0.01). (PPTX 76 kb)

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Namisaki, T., Noguchi, R., Moriya, K. et al. Beneficial effects of combined ursodeoxycholic acid and angiotensin-II type 1 receptor blocker on hepatic fibrogenesis in a rat model of nonalcoholic steatohepatitis. J Gastroenterol 51, 162–172 (2016). https://doi.org/10.1007/s00535-015-1104-x

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