Qingchangligan formula attenuates the inflammatory response to protect the liver from acute failure induced by d-galactosamine/lipopolysaccharide in mice

Abstract

Ethnopharmacological relevance: The Qingchangligan formula, a traditional Chinese medicine comprising five herbs, is useful for treatment of patients with liver failure; however, its protective and regulatory mechanisms remain elusive.

Aim of the study: To test the hypothesis that the Qingchangligan formula protects mice against acute liver failure by inhibiting liver inflammation.

Materials and methods: Acute liver failure (ALF) was induced by intraperitoneal injection of D-GalN (700mg/kg) plus LPS (10μg/kg). The Qingchangligan formula was administered to mice in three doses of 50mg/kg (on day 1, day 2, and day 3) prior to D-GalN/LPS injection by intragastric administration. The mice in different groups were sacrificed at 6h after D-GalN/LPS injection, and liver samples and blood were collected for analysis.

Results: Administration of the Qingchangligan formula not only ameliorated liver injury, as evidenced by reduced transaminase levels and well-preserved liver architecture, but also decreased the lethality in ALF mice. Moreover, in the ALF model, pretreatment with the Qingchangligan formula alleviated liver inflammation and decreased hepatocyte apoptosis. Further demonstrating the protective effects of the Qingchangligan formula, we found that pretreatment with the Qingchangligan formula reduced the expression of inflammatory cytokines by decreasing the expression of components of the mitogen-activated protein kinase (MAPK) pathway and promoting autophagy in vitro and in vivo.

Conclusions: Our findings demonstrated that the Qingchangligan formula exerts a protective effect against the pathophysiology of ALF, especially in regulating liver inflammation, and provide a rationale for using the Qingchangligan formula as a potential therapeutic strategy to ameliorate ALF.

Keywords: ALFacute liver failure; ALTalanine aminotransferase; ASTaspartate aminotransferase; Acute liver failure; Aloe-emodin (PubChem CID: 10207); Apoptosis; Atgautophagy-related gene; Autophagy; BMDMsmurine bone marrow-derived macrophages; CXCL-10chemokine (C-X-C motif) ligand-10; Caffeic acid (PubChem CID: 689043); Catalpol (PubChem CID: 91520); Chrysophanol (PubChem CID: 10208); D-GalND-galactosamine; DAPI49,6-diamino-2-phenylindole; ERKextracellular regulated protein kinase; ERendoplasmic reticulum; Emodin (PubChem CID: 3220); H&Ehematoxylin and eosin; Honokiol (PubChem CID: 72303); IL-12p40interleukin-12p40; IL-1βinterleukin-1β; Inflammation; JNKc-jun-N-terminal kinase; LPSlipopolysaccharide; MAPKmitogen-activated protein kinase; Magnolol (PubChem CID: 72300); NF-kBtranscription factor nuclear factor-kB; Naringin (PubChem CID: 442428); PAGEpolyacrylamide gel electrophoresis; PBSphosphate-buffered saline; PVDFpolyvinylidene difluoride; Physcion (PubChem CID: 10639); Qingchangligan formula; Rhein (PubChem CID: 10168); TBSTTris-buffered saline with Tween-20; TNF-αtumor necrosis factor-α; TUNELtransferase-mediated dUTP nick-end labeling; qRT-PCRquantitative reverse transcription-polymerase chain reaction.