Activation of M1 muscarinic acetylcholine receptors by proline-rich oligopeptide 7a (<EDGPIPP) from Bothrops jararaca snake venom rescues oxidative stress-induced neurotoxicity in PC12 cells

• Published in: The journal of venomous animals and toxins including tropical diseases Vol. 30; p. e20230043
• Main Authors: Alberto-Silva, Carlos, Pantaleão, Halyne Queiroz, da Silva, Brenda Rufino, da Silva, Julio Cezar Araujo, Echeverry, Marcela Bermudez
• Format: Journal Article
• Language: English
• Published: Brazil Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP) 2024; SciELO
• Subjects: Bioactive peptide; Bothrops jararaca; Neuroprotection; Proline-rich oligopeptide; TOXICOLOGY; TROPICAL MEDICINE; Bioactive peptide; Neuroprotection; Bothrops jararaca; Proline-rich oligopeptide
• ISSN: 1678-9199; 1678-9199
• DOI: 10.1590/1678-9199-JVATITD-2023-0043

The bioactive peptides derived from snake venoms of the Viperidae family species have been promising as therapeutic candidates for neuroprotection due to their ability to prevent neuronal cell loss, injury, and death. Therefore, this study aimed to evaluate the cytoprotective effects of a synthetic proline-rich oligopeptide 7a (PRO-7a; <EDGPIPP) from
snake, on oxidative stress-induced toxicity in neuronal PC12 cells and astrocyte-like C6 cells.
Both cells were pre-treated for four hours with different concentrations of PRO-7a, submitted to H
O
-induced damage for 20 h, and then the oxidative stress markers were analyzed. Also, two independent neuroprotective mechanisms were investigated: a) L-arginine metabolite generation via argininosuccinate synthetase (AsS) activity regulation to produce agmatine or polyamines with neuroprotective properties; b) M1 mAChR receptor subtype activation pathway to reduce oxidative stress and neuron injury.
PRO-7a was not cytoprotective in C6 cells, but potentiated the H
O
-induced damage to cell integrity at a concentration lower than 0.38 μM. However, PRO-7a at 1.56 µM, on the other hand, modified H
O
-induced toxicity in PC12 cells by restoring cell integrity, mitochondrial metabolism, ROS generation, and arginase indirect activity. The α-Methyl-DL-aspartic acid (MDLA) and L-N
-Nitroarginine methyl ester (L-Name), specific inhibitors of AsS and nitric oxide synthase (NOS), which catalyzes the synthesis of polyamines and NO from L-arginine, did not suppress PRO-7a-mediated cytoprotection against oxidative stress. It suggested that its mechanism is independent of the production of L-arginine metabolites with neuroprotective properties by increased AsS activity. On the other hand, the neuroprotective effect of PRO-7a was blocked in the presence of dicyclomine hydrochloride (DCH), an M1 mAChR antagonist.
For the first time, this work provides evidence that PRO-7a-induced neuroprotection seems to be mediated through M1 mAChR activation in PC12 cells, which reduces oxidative stress independently of AsS activity and L-arginine bioavailability.