Lactoferrin Protects against Methamphetamine Toxicity by Modulating Autophagy and Mitochondrial Status

Abstract

Lactoferrin (LF) was used at first as a vehicle to deliver non-soluble active compounds to the body, including the central nervous system (CNS). Nonetheless, it soon became evident that, apart from acting as a vehicle, LF itself owns active effects in the CNS. In the present study, the effects of LF are assessed both in baseline conditions, as well as to counteract methamphetamine (METH)-induced neurodegeneration by assessing cell viability, cell phenotype, mitochondrial status, and specific autophagy steps. In detail, cell integrity in baseline conditions and following METH administration was carried out by using H&E staining, Trypan blue, Fluoro Jade B, and WST-1. Western blot and immuno-fluorescence were used to assess the expression of the neurofilament marker βIII-tubulin. Mitochondria were stained using Mito Tracker Red and Green and were further detailed and quantified by using transmission electron microscopy. Autophagy markers were analyzed through immuno-fluorescence and electron microscopy. LF counteracts METH-induced degeneration. In detail, LF significantly attenuates the amount of cell loss and mitochondrial alterations produced by METH; and mitigates the dissipation of autophagy-related proteins from the autophagy compartment, which is massively induced by METH. These findings indicate a protective role of LF in the molecular mechanisms of neurodegeneration.

Alterations of Mitochondrial Structure in Methamphetamine Toxicity

“…Only a few studies have focused on mitochondrial alterations during METH toxicity [ 46 , 57 ]. When dealing with fine alterations of mitochondrial ultrastructure only a few reports are available [ 58 ]. Previously we approached such an issue by analyzing the effects of METH on mitochondrial ultrastructure in the presence of different variants of the gene expressing PINK1 , which leads to genetic parkinsonism [ 49 ].…”

Bioactive Natural Compounds for Therapeutic and Nutraceutical Applications in Neurodegeneration

“…In fact, in this study, LF Reduces METH-Induced Toxicity in PC12 Cells, whereas METH toxicity is not detected in U87MG cells, in which cell differentiation rather than neuroprotection is observed. This phenotype-dependent variability confirms that LF has antiproliferative and experimental and differentiating properties by promoting molecular pathways in ubiquitous biochemical processes that acquire different relevances depending on the specific cell type (i.e., neuroprotective or cytostatic effects) [ 17 ].…”

“…In the study reported by Ryskalin et al, LF significantly attenuates the amount of cell loss and mitochondrial alterations produced by methamphetamine (METH). Furthermore, LF can attenuate the dissipation of autophagy-related proteins, which is massively induced by METH [ 17 ]. However, the specificity of LF as a neuroprotector refers to the specific cellular phenotype.…”

Lactoferrin: neuroprotection against Parkinson's disease and secondary molecule for potential treatment

“…It is mentioned that after repeated exposure to METH in mice, a decrease in TH + cells was observed. To investigate the Lf's effect on METH-based negative mitochondrial changes and autophagy cell death, an in vitro study was performed (Ryskalin et al, 2021 ). The results indicate the Lf's positive effect on both mitochondria and the autophagy status of the cell by decreasing cellular death and relieving autophagy-related proteins from autophagy compartments.…”