The Effects of Butyrate on Induced Metabolic-Associated Fatty Liver Disease in Precision-Cut Liver Slices

Metabolic-associated fatty liver disease (MAFLD) starts with hepatic triglyceride accumulation (steatosis) and can progress to more severe stages such as non-alcoholic steatohepatitis (NASH) and even cirrhosis. Butyrate, and butyrate-producing bacteria, have been suggested to reduce liver steatosis directly and systemically by increasing liver β-oxidation. This study aimed to examine the influence of butyrate directly on the liver in an ex vivo induced MAFLD model. To maintain essential intercellular interactions, precision-cut liver slices (PCLSs) were used. These PCLSs were prepared from male C57BL/6J mice and cultured in varying concentrations of fructose, insulin, palmitic acid and oleic acid, to mimic metabolic syndrome. Dose-dependent triglyceride accumulation was measured after 24 and 48 h of incubation with the different medium compositions. PCLSs viability, as indicated by ATP content, was not affected by medium composition or the butyrate concentration used. Under induced steatotic conditions, butyrate did not prevent triglyceride accumulation. Moreover, it lowered the expression of genes encoding for fatty acid oxidation and only increased C4 related carnitines, which indicate butyrate oxidation. Nevertheless, butyrate lowered the fibrotic response of PCLSs, as shown by reduced gene expression of fibronectin, alpha-smooth muscle actin and osteopontin, and protein levels of type I collagen. These results suggest that in the liver, butyrate alone does not increase lipid β-oxidation directly but might aid in the prevention of MAFLD progression to NASH and cirrhosis.

Comment on “Circadian rhythms in the absence of the clock gene Bmal1”

Ray et al. (Reports, 14 February 2020, p. 800) recently claimed temperature-compensated, free-running mRNA oscillations in Bmal1–/– liver slices and skin fibroblasts. We reanalyzed these data and found far fewer reproducible mRNA oscillations in this genotype. We also note errors and potentially inappropriate analyses.

Nanoparticle-induced inflammation and fibrosis in ex vivo murine precision-cut liver slices and effects of nanoparticle exposure conditions

Chronic exposure and accumulation of persistent nanomaterials by cells have led to safety concerns on potential long-term effects induced by nanoparticles, including chronic inflammation and fibrosis. With this in mind, we used murine precision-cut liver tissue slices to test potential induction of inflammation and onset of fibrosis upon 72 h exposure to different nanomaterials (0–200 µg/ml). Tissue slices were chosen as an advanced ex vivo 3D model to better resemble the complexity of the in vivo tissue environment, with a focus on the liver where most nanomaterials accumulate. Effects on the onset of fibrosis and inflammation were investigated, with particular care in optimizing nanoparticle exposure conditions to tissue. Thus, we compared the effects induced on slices exposed to nanoparticles in the presence of excess free proteins (in situ), or after corona isolation. Slices exposed to daily-refreshed nanoparticle dispersions were used to test additional effects due to ageing of the dispersions. Exposure to amino-modified polystyrene nanoparticles in serum-free conditions led to strong inflammation, with stronger effects with daily-refreshed dispersions. Instead, no inflammation was observed when slices were exposed to the same nanoparticles in medium supplemented with serum to allow corona formation. Similarly, no clear signs of inflammation nor of onset of fibrosis were detected after exposure to silica, titania or carboxylated polystyrene in all conditions tested. Overall, these results show that liver slices can be used to test nanoparticle-induced inflammation in real tissue, and that the exposure conditions and ageing of the dispersions can strongly affect tissue responses to nanoparticles.

Osteoprotegerin expression in liver is induced by IL-13 through TGF-β

Backgrounds: Osteoprotegerin (OPG) is a profibrotic mediator produced by myofibroblasts under influence of transforming growth factor β (TGFβ). Its expression in experimental models of liver fibrosis correlates well with disease severity and treatment responses. The regulation of OPG in liver tissue is largely unknown and we therefore set out to elucidate which growth factors/interleukins associated with fibrosis induce OPG and through which pathways. Methods: Precision-cut liver slices of wild type and STAT6-deficient mice and 3T3 fibroblasts were used to investigate the effects of TGFβ, interleukin (IL) 13 (IL13), IL1β, and platelet-derived growth factor BB (PDGF-BB) on expression of OPG. Results: In addition to TGFβ, only IL13 and not PDGF-BB or IL1β could induce OPG expression in 3T3 fibroblasts and liver slices. This IL13-dependent induction was not shown in liver slices of STAT6-deficient mice and when wild type slices were cotreated with TGFβ receptor 1 kinase inhibitor galunisertib, STAT6 inhibitor AS1517499, or AP1 inhibitor T5224. This suggests that the OPG-inducing effect of IL13 is mediated through IL13 receptor α1-activation and subsequent STAT6-dependent upregulation of IL13 receptor α2, which in turn activates AP1 and induces production of TGFβ and subsequent production of OPG. Conclusion: We have shown that IL13 induces OPG release by liver tissue through a TGFβ-dependent pathway involving both the α1 and the α2 receptor of IL13 and transcription factors STAT6 and AP1. OPG may therefore be a novel target for the treatment liver fibrosis as it is mechanistically linked to two important regulators of fibrosis in liver, namely IL13 and TGFβ1.

In Vitro Effect of Crude Extract from Traganum Nudatum on Glucose-Uptake in Liver Slices Isolated from Westar Rats

Background: Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia resulting from defects in insulin secretion, insulin action, or both. There are many classes of drugs used for treatment, and these include insulin sensitizers, insulin secretagogues, and agents that delay the absorption of carbohydrates from the bowel. This study intends to investigate the effect of crude extract from a plant from South Algeria Traganum nudatum (Chenopodiaceae) on glucose uptake in liver slices isolated from Wistar rats. Methods: The liver slices were incubated for 90 min at 37° in normoglycaemic (1g/l of glucose) and hyperglycaemic (3g/l of glucose) KRBA Krebs Ringer Bicarbonate Albumin 4% media using 24 well-polyethylene plates. In each, well different concentrations of insulin (10, 50 and 100µU/ml) and hydromethanolic crude extract (100, 200 and 500µg/ml) were added. After every 30 minutes, aliquots of the culture media were assayed for the determination of glucose left. Results: Tests showed that the glucose left after 90 minutes in the media which contained insulin at 100µg/ml was the lowest (0.44 and 1.41 )g/l in the normo and hyperglycaemic media respectively, which reflect that insulin at this concentration was the most effective on the stimulation of glucose uptake. The extract had the highest effect at 500µg/ml, the concentrations of glucose left after 90 minutes of incubation were found to be (0.38 and 1.31)g/l in the normoglycaemic and hyperglycaemic media respectively. Conclusion: From the obtained results, it can be concluded that our extract seems to have an insulin-like effect on glucose uptake in liver slices isolated from Wistar rats.