Characterization and neurotherapeutic evaluation of venom polypeptides identified from Vespa magnifica: The role of Mastoparan-M in Parkinson’s disease intervention

Abstract

Ethnopharmacological relevance

Parkinson’s disease (PD) is a common neurodegenerative disorder in the elderly, characterized by the loss of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies. Hufeng Jiu from Vespa magnifica Smith, a traditional remedy used by the Chinese Jingpo minority, is documented in the Pharmacopoeia of China (2020) for treating rheumatic arthritis. Notably, recent research suggests that components of wasp venom (WV) from Vespa magnifica Smith, particularly polypeptides such as Mastoparan-M (Mast-M) and Vespakinin-M, may have potential therapeutic effects for neurological disorders. However, the specific polypeptide components of WV and their therapeutic effects on PD models remain insufficiently understood.

Aim of the study

This study aims to characterize the neuroactive polypeptides in Vespa magnifica Smith venom and investigate the therapeutic potential of Mast-M for PD.

Materials and methods

Neuroactive polypeptides in WV were identified using LC/MS, and Mast-M derived from venom of Vespa magnifica Smith was verified with HPLC. The neuroprotective effects of WV and its peptides were assessed using the CCK-8 assay in 1-methyl-4- phenylpyridinium (MPP⁺)-induced SH-SY5Y human neuroblastoma cells. Mast-M was identified as a potent antagonist against MPP⁺-induced neurotoxicity. The toxicity, hemolytic activity, and blood-brain-barrier (BBB) permeability of Mast-M were evaluated in mice, and its therapeutic effects were assessed in an MPTP-induced PD mouse model, focusing on motor function and tyrosine hydroxylase (TH) levels. Additionally, Mast-M’s impact on mitochondrial membrane potential (MMP), autophagy, and the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signling pathway was investigated.

Results

A total of 1007 peptides were identified in the WV, including 187 UniProtKB unreviewed, with 185 predicted to be BBB-permeability. Our results show that Mast-M exhibits a time-dependent distribution in mice, initially localizing in the peritoneal region and subsequently accumulating in the brain, liver, and kidney. Cellular uptake studies reveal that Mast-M penetrates cell membranes and accumulates intracellularly over time. In the MPP⁺-induced neurotoxicity model using SH-SY5Y cells, Mast-M significantly enhances cell viability and MMP. In vivo safety assessments indicate that Mast-M is well-tolerated at doses up to 100 μg/kg, with no significant toxicological effects observed. However, higher doses induce hepatic distress, necessitating dose optimization. Hemolysis was absent at concentrations ≤37 μg/mL, with an EC₅₀ for hemolytic activity of 197 μg/mL. In MPTP-induced PD models, Mast-M partially ameliorates motor deficits and preserves TH expression in dopaminergic neurons, supporting its neuroprotective role. Mechanistically, Mast-M activates autophagic pathways, as evidenced by the upregulation of autophagy-related protein LC3 in MPP⁺-challenged SH-SY5Y cells. Furthermore, Mast-M promotes mitophagy and mitochondrial biogenesis, modulating the AMPK/mTOR signaling axis to facilitate mitochondrial turnover.

Conclusion

Mast-M emerges as a promising therapeutic candidate for PD, capable of crossing the BBB, enhancing autophagy, and providing neuroprotection in PD models. Further studies are warranted to optimize dosing and elucidate its full therapeutic potential.