Induction of chronic cholestasis without liver cirrhosis - Creation of an animal model

Abstract

AIMTo analyze time intervals of inflammation and regeneration in a cholestatic rat liver model.METHODSIn 36 Lewis rats, divided into six groups of 6 animals (postoperative observation periods: 1, 2, 3, 4, 6, 8 wk), the main bile duct was ligated with two ligatures and observed for the periods mentioned above. For laboratory evaluation, cholestasis parameters (bilirubin, γ-GT), liver cell parameters (ASAT, ALAT) and liver synthesis parameters (quick, albumin) were determined. For histological analysis, HE, EvG, ASDCL and HMGB-1 stainings were performed. Furthermore, we used the mRNA of IL-33, GADD45a and p-21 for analyzing cellular stress and regeneration in cholestatic rats.RESULTSIn chemical laboratory and histological evaluation, a distinction between acute and chronic cholestatic liver injury with identification of inflammation and regeneration could be demonstrated by an increase in cholestasis (bilirubin: 1-wk group, 156.83 ± 34.12 μmol/L, P = 0.004) and liver cell parameters (ASAT: 2-wk group, 2.1 ± 2.19 μmol/L.s, P = 0.03; ALAT: 2-wk group, 1.03 ± 0.38 μmol/L.s, P = 0.03) after bile duct ligation (BDL). Histological evaluation showed an increase of bile ducts per portal field (3-wk group, 48 ± 6.13, P = 0.004) during the first four weeks after bile duct ligation. In addition to inflammation, which is an expression of acute cholestasis, there was an increase of necrotic areas in the histological sections (2-wk group, 1.38% ± 2.28% per slide, P = 0.002). Furthermore, the inflammation could be verified by ASDCL (4-wk group, 22 ± 5.93 positive cells per portal field, P = 0.041) and HMGB-1 [2-wk group, 13 ± 8.18 positive cells per field of view (FoV), P = 0.065] staining. Therefore, in summary of the laboratory evaluation and histological studies, acute cholestasis could be found during the first four weeks after bile duct ligation. Subsequently, the described parameters declined so that chronic cholestasis could be assumed. For quantification of secondary biliary cirrhosis, eosin staining was performed, which did not reveal any signs of liver remodeling, thus precluding the development of a chronic cholestasis model. Additionally, to establish the chronic cholestasis model, we evaluated liver regeneration capacity through measurements of IL-33, p-21 and GADD45a mRNA.CONCLUSIONWe created a chronic cholestasis model. The point of inflammatory and regenerative balance was reached after four weeks. This finding should be used for experimental approaches dealing with chronic cholestatic liver damage.

Trehalose reduces bone loss in experimental biliary cirrhosis rats via ERK phosphorylation regulation by enhancing autophagosome formation

“…Similarly, some studies demonstrated reduced bone formation and increased bone resorption in BDL male rats, 27,28 which is in accordance with the PBC patients 30 . The BDL male rat model used in the present study was simple and effective, as described previously 27,28,31 . Many scholars considered that cholestasis is one of the major risk factors of impaired bone formation 32 .…”

Hepatotoxicity of a Cannabidiol-Rich Cannabis Extract in the Mouse Model

“…Previous studies have demonstrated that down-regulation of Cdkn1a , also known as p21 , stimulates liver regeneration, while overexpression inhibits this process [36,37,38]. Furthermore, a recent report of Cdkn1a up-regulation resulting from acute cholestatic injury was proposed as a biomarker for impaired liver regeneration [39]. Elevated total bilirubin in conjunction with up-regulation of Cdkn1a and a number of other gene-markers of cholestatic liver injury (i.e., Abcb1a , Abcc2 , Abcc3 , Atp8b1 , and Rdx ) and down-regulation of fatty acid metabolism-related genes ( Car3 , Fabp1 , and Ppara ) observed in our study, suggest that CBD-induced liver injury may be cholestatic, though ALP and GGT were not elevated.…”

Quercetin mitigates liver injury in a rat model of liver cholestasis

“…Therefore, it is highly possible that quercetin may be effective in prohibiting oxidative stress induced liver injury in chronic cholestasis. Thus, the present study was planned to investigate the possible protective effect and the related mechanisms of quercetin against cholestasis induced by bile duct ligation in rats, a well settled experimental model of cholestasis that initiates a complex cascade of pathological events similar to that of human [12].…”