Development of an alternative zebrafish model for drug‐induced intestinal toxicity

Abstract

An evaluation of intestinal toxicity is important because the mucosal lining of the gastrointestinal tract is the first barrier for oral xenobiotics. Until now, a rat model has been recommended as the standard intestinal toxicity model and the Caco-2 cell line, originated from a human colon adenocarcinoma, has been used as an alternative to this model, but there are limitations regarding cost-effectiveness and the need for mimicry of the human system. In this study, we investigated whether zebrafish could be a valid alternative to rats and Caco-2 cells as an intestinal toxicity model. We focused on intestinal gene expression of cytochrome P450 3A65, oxidative stress, apoptosis, inflammation, and intestinal function. Reverse transcription-quantitative polymerase chain reaction analysis was conducted using three models: zebrafish, Sprague-Dawley rats and Caco-2 cells, and the transcript levels and patterns of indicator genes were analyzed in conjunction with histopathological changes. Our results suggested that representative intestinal toxicants, indomethacin, diclofenac and methotrexate, induced significant transcript level changes in marker genes such as CYP3A, inducible nitric oxide synthase, heme oxygenase 1, superoxide dismutase 1, glutathione peroxidase 1, BCL2 associated X, B-cell lymphoma 2, caspase 9, tumor protein p53, nuclear factor-κB, interleukin-1β, tumor necrosis factor-alphaα and toll-like receptor 2 in the zebrafish model as in the rat and Caco-2 cells models. These results suggest that zebrafish model is sufficiently worth developing as an intestinal toxicity model that can replace or compensate the rat model or Caco-2 cell model.

Titanium dioxide nanoparticles promote histopathological and genotoxic effects in Danio rerio after acute and chronic exposures

“…In toxicological studies, the intestine is a relevant organ because of being the first physiological barrier for orally ingested chemicals that can be potentially toxic (RYU et al, 2018). In the present study, there was an increase in the thickness of intestinal villi, also called as hyperplasia, in 20 and 40 mg L -1 of TiO2-NPs in the acute and chronic exposures.…”

Drug-induced gene expression profile changes in relation to intestinal toxicity: State-of-the-art and new approaches

“…Overall, the majority of the selected genes were upregulated in the three toxicity models after exposure to one of the three compounds, namely CYP3A (metabolism); iNOS, Hmox1, Sod1 and Gpx1 (oxidative stress); Bax, Bcl-2, Casp9 and p53 (apoptosis); IL-1β, TNF-α and TIr2 (inflammation). Expression levels of NF-kB and Occludin (intestinal tight junction) were decreased after exposure to the drugs [78]. Altogether, this study shows putative genes that are affected by NSAIDs exposure that can be part of the transcriptomic signature of these NSAIDs.…”

“…Only one in vivo study was performed to evaluate the whole genome transcriptomic responses associated with NSAIDs intestine toxicity so far [78]. In this study, Ryu et al compared 2 animal models (zebrafish and rat) with the human colonic adenocarcinoma cell line Caco-2, aiming at developing an alternative animal model (zebrafish) to study intestinal toxicity induced by drugs.…”

“…In this study, Ryu et al compared 2 animal models (zebrafish and rat) with the human colonic adenocarcinoma cell line Caco-2, aiming at developing an alternative animal model (zebrafish) to study intestinal toxicity induced by drugs. All three models were exposed to indomethacin, diclofenac and methotrexate [78]. Specific biomarkers were selected to evaluate the effects of those drugs, including biomarkers of metabolism, oxidative stress, apoptosis, inflammation and intestinal structural changes, and ultimately compare the results between the different models.…”

Effects of MP Polyethylene Microparticles on Microbiome and Inflammatory Response of Larval Zebrafish

“…Nine genes were selected in order to examine the inflammatory response of larval zebrafish. Primer sets were obtained from previous publications or custom designed with the use of Primer3 software ( Table 1 , [ 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 ]). Primer specificity was confirmed by examining the top e-value in BLASTn (NCBI) and single product amplification confirmed by melt analysis of all reactions.…”