Protective effect of calpain inhibitors against manganese-induced toxicity in rats

Development of manganism is a major complication of manganese exposure in which neurological dysfunction is linked to accumulation of metal in the brain. Current therapies do not prevent progression of the disease. Therefore, development of effective therapeutic strategies for treatment of manganism is of utmost importance. Since the hyperactivation of calpain family proteases in CNS during manganism in an animal model is observed, we assumed that inhibition of calpains can suppress the development of Mn-induced neurological disturbances. The goal of this study is to delineate protective effect and the mechanism of neuroprotection of calpain inhibitor in rat model of Mn-induced neurological symptoms. Using the Gait analysis test, we found that chronic intranasal administration of the calpain inhibitor Cast (184-210) (peptide, which is corresponding to the 184-210 amino acid of the endogenous inhibitor of calpains-human calpastatin) to Mn-treated rats contributed to a significant decrease in the severity of gait disorders, although it did not lead to a decrease in the Mn deposition in the striatum and hippocampus. Accordingly to the results of PCR-RT, this effect was accompanied by a partial reduction in the content of neuro-inflammatory markers (IL-1β, TNF-α, NFκB mRNA in the hippocampus and, additionally, IBA-1 mRNA in the striatum), as well as normalization of the content of dopamine and its metabolites in the hippocampus and striatum, which was assessed by HPLC. In striatum cells, the application of Cast (184-210) also led to a significant increase in the production of tyrosine hydroxylase, which was analyzed by immunoblotting method. These findings suggest that calpain inhibitors may be a valid therapeutic agent in manganism.