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Toxicology. 2018 Jan 15;393:150-159. doi: 10.1016/j.tox.2017.11.010. Epub 2017 Nov 7.

The high-production volume fungicide pyraclostrobin induces triglyceride accumulation associated with mitochondrial dysfunction, and promotes adipocyte differentiation independent of PPARγ activation, in 3T3-L1 cells.

Author information

1
Nicholas School of the Environment, Box 90328, Duke University, Durham, NC, 27708, USA. Electronic address: anthony.luz@duke.edu.
2
Nicholas School of the Environment, Box 90328, Duke University, Durham, NC, 27708, USA. Electronic address: christopher.kassotis@duke.edu.
3
Nicholas School of the Environment, Box 90328, Duke University, Durham, NC, 27708, USA. Electronic address: heather.stapleton@duke.edu.
4
Nicholas School of the Environment, Box 90328, Duke University, Durham, NC, 27708, USA. Electronic address: joel.meyer@duke.edu.

Abstract

Pyraclostrobin is one of the most heavily used fungicides, and has been detected on a variety of produce, suggesting human exposure occurs regularly. Recently, pyraclostrobin exposure has been linked to a variety of toxic effects, including neurodegeneration and triglyceride (TG) accumulation. As pyraclostrobin inhibits electron transport chain complex III, and as mitochondrial dysfunction is associated with metabolic syndrome (cardiovascular disease, type II diabetes, obesity), we designed experiments to test the hypothesis that mitochondrial dysfunction underlies its adipogenic activity. 3T3-L1 cells were differentiated according to standard protocols in the presence of pyraclostrobin, resulting in TG accumulation. However, TG accumulation occurred without activation of the peroxisome proliferator activated nuclear receptor gamma (PPARγ), the canonical pathway mediating adipogenesis. Furthermore, cells failed to express many markers of adipogenesis (PPARγ, lpl, CEBPα), while co-exposure to pyraclostrobin and two different PPARγ antagonists (GW9662, T0070907) failed to mitigate TG accumulation, suggesting TG accumulation occurred through a PPARγ-independent mechanism. Instead, pyraclostrobin reduced steady-state ATP, mitochondrial membrane potential, basal mitochondrial respiration, ATP-linked respiration, and spare respiratory capacity, demonstrating mitochondrial dysfunction, while reduced expression of genes involved in glucose transport (Glut-4), glycolysis (Pkm, Pfkl, Pfkm), fatty acid oxidation (Cpt-1b), and lipogenesis (Fasn, Acacα, Acacβ) further suggested a disruption of metabolism. Finally, inhibition of cAMP responsive element binding protein (CREB), a PPARγ coactivator, partially mitigated pyraclostrobin-induced TG accumulation, suggesting TG accumulation is occurring through a CREB-driven mechanism. In contrast, rosiglitazone, a known PPARγ agonist, induced TG accumulation in a PPARγ-dependent manner and enhanced mitochondrial function. Collectively, these results suggest pyraclostrobin-induced mitochondrial dysfunction inhibits lipid homeostasis, resulting in TG accumulation. Exposures that disrupt mitochondrial function may have the potential to contribute to the rising incidence of metabolic syndrome, and thus more research is needed to understand the human health impact of pyraclostrobin exposure.

KEYWORDS:

Adipogenesis; Fungicide; Metabolic disruption; Mitochondrial toxicity; Pyraclostrobin

PMID:
29127035
PMCID:
PMC5726929
DOI:
10.1016/j.tox.2017.11.010
[Indexed for MEDLINE]
Free PMC Article

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