Precision-cut liver slices as a model for assess hepatic cellular response of chitosan–glutathione nanoparticles on cultures treated with zilpaterol and clenbuterol

Abstract

Zilpaterol and clenbuterol are two β-adrenergic agonist drugs used in animal production. Both drugs have anabolic effects with advantages on carcass yield. Meanwhile, zilpaterol is approved for animal feed in authorized countries. Clenbuterol is a banned substance due to the risk of toxicity; however, it is still being used in unknown dose levels in many farm species. Therefore, the use and abuse of these substances should be closely monitored, considering the clenbuterol ability and the not proved yet of zilpaterol to produce reactive oxygen and nitrogen species. Regarding glutathione which is the main intracellular antioxidant plays detoxification functions on liver metabolism; in this work, it is our interest to know the capacity of chitosan–glutathione nanoparticles (CS/GSH-NP) as a complementary source of exogenous GSH to modify the oxide-reduction status on bovine precision-cut liver slice cultures (PCLS) exposed to clenbuterol and zilpaterol. A single drug assay was performed in first instance by adding clenbuterol, zilpaterol, chitosan nanoparticles (CS-NP), and CS/GSH-NP. Then combinate drug assay was carried out by testing clenbuterol and zilpaterol combined with CS-NP or CS/GSH-NP. The results showed that both β-adrenergic agonists modify in a dose-dependent manner in oxide-reduction response through ROS generation. The activity or content of glutathione peroxidase activity, intracellular GSH, gamma glutamyl-transpeptidase, aspartate aminotrasnferase and alanine aminotrasnferase were modified. The exogenous GSH delivered by nanoparticles could be used to modulate these markers.

Keywords:

• Clenbuterol
• zilpaterol
• chitosan–glutathione nanoparticles
• precision-cut liver slices
• redox response

Acknowledgments

The authors thank Luis Alberto Parra Oaxaca for his technical support on the supply of liver samples.

Compliance with ethical standards

This study used bovine liver samples obtained from animals slaughtered in the municipal slaughterhouse of Zumpango, México State, México. The animals correspond to those destined for human consumption. All the procedures used were according to the local care legislation NOM-033-SAG/ZOO-2014 and the suggested methods to slaughter domestic and wild animals (DOF 2015).

Disclosure statement

The authors declare no conflict of interest.

Additional information

Funding

This work was supported by the National University Autonomous of México DGAPA-PAP IIT under Grants: IN214321 and IT201620 and National Council of Science and Technology doctoral fellowship 289464.