ABSTRACT Background High intakes of fructose are associated with metabolic diseases, including hypertriglyceridemia and intestinal tumor growth. Although small intestinal epithelia consist of many different cell types, express lipogenic genes, and convert dietary fructose to fatty acids, there is no information on the identity of the cell type(s) mediating this conversion and on the effects of fructose on lipogenic gene expression. Objectives We hypothesized that fructose regulates the intestinal expression of genes involved in lipid and apolipoprotein synthesis, that regulation depends on the fructose transporter solute carrier family 2 member a5 [Slc2a5 (glucose transporter 5)] and on ketohexokinase (Khk), and that regulation occurs only in enterocytes. Methods We compared lipogenic gene expression among different organs from wild-type adult male C57BL mice consuming a standard vivarium nonpurified diet. We then gavaged twice daily for 2.5 d fructose or glucose solutions (15%, 0.3 mL per mouse) into wild-type, Slc2a5-knockout (KO), and Khk-KO mice with free access to the nonpurified diet and determined expression of representative lipogenic genes. Finally, from mice fed the nonpurified diet, we made organoids highly enriched in enterocyte, goblet, Paneth, or stem cells and then incubated them overnight in 10 mM fructose or glucose. Results Most lipogenic genes were significantly expressed in the intestine relative to the kidney, liver, lung, and skeletal muscle. In vivo expression of Srebf1, Acaca, Fasn, Scd1, Dgat1, Gk, Apoa4, and Apob mRNA and of Scd1 protein increased (P < 0.05) by 3- to 20-fold in wild-type, but not in Slc2a5-KO and Khk-KO, mice gavaged with fructose. In vitro, Slc2a5- and Khk-dependent, fructose-induced increases, which ranged from 1.5- to 4-fold (P < 0.05), in mRNA concentrations of all these genes were observed only in organoids enriched in enterocytes. Conclusions Fructose specifically stimulates expression of mouse small intestinal genes for lipid and apolipoprotein synthesis. Secretory and stem cells seem incapable of transport- and metabolism-dependent lipogenesis, occurring only in absorptive enterocytes.
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Early enteric and hepatic responses to ingestion of polystyrene nanospheres from water in C57BL/6 mice
Drinking water is one of numerous sources of human exposure to microscale and nanoscale plastic particles. Here, using a mouse model, we tested enteric and hepatic cellular responses to nanoplastic ingestion. At 1.5 or 25.5 h after an oral dose of 70 mg polystyrene nanospheres (PSNS)/kg (nominal diameters of 20 and 200 nm) in aqueous suspension female mice exhibit no overt signs of toxicity. Routine histopathology on small intestine and liver reveals no acute toxicity. Immunohistochemistry detects an increase in the number of enterocytes with cleaved caspase-3 (active form) after PSNS exposure ( p ≤ 0.05) indicating progression toward lytic cell death via a proinflammatory pathway. This is not evident in liver after PSNS exposure. Transmission electron microscopy detects lytic cell death in enterocytes at 25.5 h after 200 nm PSNS exposure. Putative endosomes in liver appear to sequester 20 and 200 nm particles 25.5 h after exposure. Both 20 and 200 nm PSNS appear in putative perinuclear autolysosomes 25.5 h after treatment. No significant changes in gene expression in the small intestine or liver 25.5 h were observed after dosing, but there was a trend toward altered expression of cyp1b1 in the liver. Analysis of the fecal microbiome shows loss of diversity after exposure to both 20 and 200 nm particles after 25.5 h. Taken together, these results suggest risk from ingestion of nanoscale plastic particles from drinking water, which deserves systematic investigation.
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- Award ID(s):
- 1935028
- PAR ID:
- 10357885
- Date Published:
- Journal Name:
- Frontiers in Water
- Volume:
- 4
- ISSN:
- 2624-9375
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Increasing use of nanoscale lithium cobalt oxide (LCO) particles in nanotechnologies, including catalysis and energy storage, raises concerns over their release into the environment and subsequent biological impact. Here we study the impact of LCO nanosheets on trout gill epithelial cells – model cells for environmental exposures – by global transcriptomics using RNA-Seq. We identify molecular processes impacted by subtoxic and toxic nanoparticle (NP) doses as well as dissolved Li + and Co 2+ ions. We found that the ions, at concentrations released from the toxic NP dose, did not impact cell viability and downregulated the expression of few genes following 24 h exposure, which recovered to normal levels at 48 h. In contrast, the toxic NP dose upregulated the expression of over 1000 genes at each time point, indicating the intact NPs are responsible for perturbing gene expression and toxicity. Importantly, the subtoxic NP dose, despite having no impact on cell viability, upregulated the expression of over 500 genes at 24 h, and 150 genes at 48 h. Clustering analysis showed distinct gene expression profiles induced by the toxic and subtoxic NP doses, and functional enrichment identified pathways with distinct patterns of regulations. The impacted pathways fell into four main functional categories: metabolic and energy related processes, oxygen and hypoxia related processes, membrane binding and internalization processes, and developmental processes. Together, our observations indicate that LCO NP toxicity originates from the intact NP, not the dissolved ions, and even subtoxic NP dose impacts multiple pathways critical to the normal function of the cell.more » « less
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.3389/frwa.2022.925781
