Obese diet-induced mouse models of nonalcoholic steatohepatitis-tracking disease by liver biopsy

Abstract

Author contributions: Kristiansen MNB, Veidal SS, Rigbolt KTG, Tølbøl KS and Feigh M performed the experiments and analyzed the data; Rigbolt KTG performed the molecular investigations; Kristiansen MNB and Veidal SS performed the histological analysis; Veidal SS, Rigbolt KTG, Roth JD, Jelsing J, Vrang N and Feigh M designed and coordinated the research; Kristiansen MNB, Veidal SS, Rigbolt KTG, Tølbøl KS, Roth JD, Jelsing J, Vrang N and Feigh M wrote the paper.Institutional review board statement: This study includes no data or material from patients. We confirm that all of the required permissions for this study were obtained from our local authorities as mentioned in the Institutional animal care and use committee statement.Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Danish Committee for animal research and covered by a personal license for Jacob Jelsing (2013-15-2934-00784). All of the institutional and national guidelines for the care and use of laboratory animals were followed.Conflict-of-interest statement: There are no patents, products in development or marked products to declare. Abstract AIM: To characterize development of diet-induced nonalcoholic steatohepatitis (NASH) by performing liver biopsy in wild-type and genetically obese mice. METHODS:Male wild-type C57BL/6J (C57) mice (DIO-NASH) and male Lep ob /Lep ob (ob /ob ) mice (ob /ob -NASH) were maintained on a diet high in trans-fat (40%), fructose (22%) and cholesterol (2%) for 26 and 12 wk, respectively. A normal chow diet served as control in C57 mice (lean chow) and ob /ob mice (ob /ob chow).After the diet-induction period, mice were liver biopsied and a blinded histological assessment of steatosis and fibrosis was conducted. Mice were then stratified into groups counterbalanced for steatosis score and fibrosis stage and continued on diet and to receive daily PO dosing of vehicle for 8 wk. Global gene expression in liver tissue was assessed by RNA sequencing and bioinformatics. Metabolic parameters, plasma liver enzymes and lipids (total cholesterol, triglycerides) as well as hepatic lipids and collagen content were measured by biochemical analysis. Non-alcoholic fatty liver disease activity score (NAS) (steatosis/inflammation/ballooning Obese diet-induced mouse models of nonalcoholic steatohepatitis-tracking disease by liver biopsy Basic Study ORIGINAL ARTICLEdegeneration) and fibrosis were scored. Steatosis and fibrosis were also quantified using percent fractional area. RESULTS: Diet-induction for 26 and 12 wk in DIO-NASH and ob /ob -NASH mice, respectively, elicited progressive metabolic perturbations characterized by increased adiposity, total cholesterol and elevated plasma liver enzymes. The diet also induced clear histological features of NASH including hepatosteatosis and fibrosis. Overall, the metabolic NASH phenotype was more pronounced in ob /ob -NASH vs DIO-NASH mice. During the eight week repeated vehicle dosing period, the metabolic phenotype was sustai...

Non‐alcoholic fatty liver disease alters expression of genes governing hepatic nitrogen conversion

“…Liver transcriptome analysis was performed by RNA sequencing on RNA extracted from liver biopsies as previously described . All biopsies were of high RNA quality with integrity number ≥7.5.…”

“…Liver transcriptome analysis was performed by RNA sequencing on RNA extracted from liver biopsies as previously described. 10,16 All biopsies were of high RNA quality with integrity number ≥7.5. • In contrast, the gene expression of the glutamine synthetase located perivenously further down the sinusoid was strongly upregulated suggesting a possible adaptive mechanism for ammonia scavenging.…”

A Therapeutic Silencing RNA Targeting Hepatocyte TAZ Prevents and Reverses Fibrosis in Nonalcoholic Steatohepatitis in Mice

“…Thus GalNAc‐siTAZ efficiently and specifically lowers liver hepatocyte TAZ in NASH diet‐fed mice. We also tested TAZ silencing efficiency after one injection of GalNAc‐siTAZ in mice fed a high‐fat/cholesterol/fructose diet enriched in transfatty acids (amylin liver NASH [AMLN] diet) for 61 weeks, which induces substantial fibrotic NASH after long‐term feeding . As with the NASH model above, GalNAc‐siTAZ was very effective at silencing liver TAZ protein and Wwtr1 mRNA (Supporting Fig.…”

NAFLD Preclinical Models: More than a Handful, Less of a Concern?

“…This model covers a high-fat/high-fructose (40%/22%) diet containing trans-fatty acids (~18%) and high-cholesterol (2%), thereby better resembling the Western-type diet and subsequent development of NASH features. Remarkably, only after 26-30 weeks of feeding an AMLN diet, wild-type C57BL/6 developed marked steatosis, moderate lobular inflammation and hepatocellular ballooning [29]. However, when obese leptin-deficient ob/ob mice were fed a similar AMLN diet for 12 weeks, mice displayed an accelerated and more pronounced metabolic NASH phenotype as compared to wild-type C57BL/6 [29].…”