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Statins are the most prescribed lipid-lowering agents worldwide. Their use is generally safe, although muscular toxicity occurs in 1 in 10.000 patients. In this study, we explored the role of the endocannabinoid system (ECS) during muscle toxicity induced by simvastatin. In murine C2C12 myoblasts exposed to simvastatin (30 µM), we found that the levels of the endocannabinoids 2-AG and AEA as well the expression of specific miRNAs (mostly miR-152) targeting the endocannabinoid CB1 gene were increased. Rimonabant, a selective CB1 antagonist, exacerbated simvastatin-induced toxicity in myoblasts, while the opposite effect was observed with GAT211, a CB1-positive allosteric modulator. In antagomiR-152-transfected myoblasts, simvastatin toxicity was prevented along with the rescue of CB1 expression. Notably, similar alterations were found in skeletal muscles of C57BL/6J mice treated with simvastatin 20 mg Kg-1 and in primary human myoblasts. In sum, we identified the ECS as a novel mechanism participating in statin-induced myopathy.

N-oleoylglycine (OlGly) is a lipid mediator that belongs to the expanded version of the endocannabinoid (eCB) system, the endocannabinoidome (eCBome), which has recently gained increasing attention from the scientific community for its protective effects in a mouse model of mild traumatic brain injury. However, the effects of OlGly on cellular models of Parkinson’s disease (PD) have not yet been investigated, whilst other lipoaminoacids have been reported to have beneficial effects. Moreover, the protective effects of OlGly seem to be mediated by direct activation of proliferator-activated receptor alpha (PPARα), which has already been investigated as a therapeutic target for PD. Therefore, this study aims to investigate the possible protective effects of OlGly in an in vitro model obtained by treating the neuroblastoma cell line, SH-SY5Y (both differentiated and not) with 1-methyl-4-phenyl-pyridinium (MPP+), which mimics some cellular aspects of a PD-like phenotype, in the presence or absence of the PPARα antagonist, GW6471. Our data show that MPP+ increases mRNA levels of PPARα in both non differentiated and differentiated cells. Using assays to assess cell metabolic activity, cell proliferation, and pro-inflammatory markers, we observed that OlGly (1 nM), both as treatment (1 h) and pre-treatment (4 h), is able to protect against neuronal damage induced by 24 h MPP+ exposure through PPARα. Moreover, using a targeted lipidomics approach, we demonstrate that OlGly exerts its effects also through the modulation of the eCBome. Finally, treatment with OlGly was able also to reduce increased IL-1β induced by MPP+ in differentiated cells. In conclusion, our results suggest that OlGly could be a promising therapeutic agent for the treatment of MPP+-induced neurotoxicity.

Background and Purpose Statins are the most common prescribed lipid-lowering agents worldwide. Although their use is generally safe, statins cause muscle disorders in 1 in 10.000 people. In this study, we investigated the role of the endocannabinoid system (ECS) in statin-induced myotoxicity. Experimental Approach ECS activity was evaluated in murine C2C12 and primary human myoblasts exposed to simvastatin (30 µM) as well as in skeletal muscles of C57BL/6J mice receiving oral simvastatin (20 mg kg−1) for three weeks using LC-MS, qPCR and western blot analyses. Bioinformatic, genetic and pharmacological tools followed by qPCR and FACS analyses, were used to explore the role of the cannabinoid CB1 receptor. Key Results In C2C12 cells, simvastatin caused dysregulation in levels of the two major endocannabinoids 2-AG and AEA as well as upregulation of miRNAs, mostly miR-152, which repress the expression of the endocannabinoid receptor, CB1. Notably, simvastatin-induced toxicity in myoblasts was exacerbated in the presence of the selective CB1 antagonist rimonabant, and slightly attenuated by the CB1 positive allosteric modulator GAT211. In antagomiR-152-transfected C2C12 myoblasts, simvastatin toxicity was prevented along with the rescue of CB1 expression. Simvastatin caused similar alterations in skeletal muscles of C57BL/6J mice, and most importantly, in primary human myoblasts. Conclusions and Implications In both murine and human skeletal muscle cells, simvastatin induces changes in ECS activity also caused by dysregulation of specific miRNAs inducing repression of CB1 expression. In sum, we propose a novel mechanism to be targeted for preventing statins-induced myopathy.

Chromium (VI) is the most dangerous oxidation state among the stable forms of chromium. In this work, we evaluated the effect of exposing Mytilus galloprovincialis for 24 h to 1, 10, and 100 nM chromium (VI) on the properties of Protamine-like (PLs) and their gene levels in the gonads. Specifically, we analyzed, by AU-PAGE and SDS-PAGE, PLs extracted from unexposed and exposed mussels. In addition, via EMSA, we evaluated the ability of PLs to bind DNA and also verified their potential to protect DNA from oxidative damage. Finally, we assessed possible alterations in gonadal expression of mt10, hsp70, and genes encoding for PLs-II/PL-IV and PL-III. We found that for all experimental approaches the most relevant alterations occurred after exposure to 1 nM Cr(VI). In particular, a comigration of PL-II with PL-III was observed by SDS-PAGE; and a reduced ability of PLs to bind and protect DNA from oxidative damage was recorded. This dose of chromium (VI) exposure was also the one that produced the greatest alterations in the expression of both mt10 and PL-II/PL-IV encoding genes. All of these changes suggest that this dose of chromium (VI) exposure could affect the reproductive health of Mytilus galloprovincialis.

In our previous work, we reported alterations in protamines/histones ratio, in DNA binding of these proteins and their involvement in DNA oxidative damage in 84% of the young men living in the Land of Fires. In the present work, we extended our findings, evaluating any alterations in spermatozoa of a family case, a father and son, living in this area, to also give a first look at the possibility of transgenerational inherited effects of environmental contaminants on the molecular alterations of sperm nuclear basic proteins (SNBP), DNA and semen parameters. In the father and son, we found a diverse excess of copper and chromium in the semen, different alterations in SNBP content and low DNA binding affinity of these proteins. In addition, DNA damage, in the presence of CuCl2 and H2O2, increased by adding both the father and son SNBP. Interestingly, son SNBP, unlike his father, showed an unstable DNA binding and were able to produce DNA damage even without external addition of CuCl2, in line with a lower seminal antioxidant activity than the father. The peculiarity of some characteristics of son semen could be a basis for possible future studies on transgenerational effects of pollutants on fertility.