Mastoparan M

Mastoparan M, a tetradecapeptide toxin (INKAIAALAKKLL-NH2) from hornet venom and its D-form mastoparan M were synthesized chemically. All D- and L-mastoparan M forms were found to adopt 28% alpha-helical structures in a 30% trifluroethanol solution as shown by the circular dichroism spectrum. All-D mastoparan M caused 3H-thymidine release from labeled bacterial cells after incubation for 1 h and complete cell lysis by 4 h. Both L- and D-mastoparan M showed strong activity against gram-positive and gram-negative bacteria. All-D mastoparan M showed 2-fold higher antibacterial activity than L-mastoparan M. The effects of all-D mastoparan M on the surface morphology of Staphylococcus aureus (ATCC29213) and Escherichia coli (ATCC25922) were studied by scanning-beam electron microscopy. Blast-like bleb extrusions on the surface of some S. aureus and swellings on the end of E. coli were seen after culture with all-D mastoparan M. These findings indicated the all-D mastoparan M could kill bacteria by disrupting cells.

Objectives: Global cerebral ischemia (GCI), a consequence of cardiac arrest (CA), can significantly damage the neurons located in the vulnerable hippocampus CA1 areas. Clinically, neurological injury after CA contributes to death in most patients. Mastoparan-M extracted from Vespa magnifica (Smith) can be used to treat major neurological disorders. Hence, this study aimed to assess the effects of Mastoparan-M on GCI. Materials and methods: To evaluate the neurotoxicity and neuroprotective effect of Mastoparan-M, the CCK8 and Annexin V-FITC/PI apoptosis assays were first performed in hippocampal HT22 neuronal cells in vitro. Then, Pulsinelli's 4-vascular occlusion model was constructed in rats. After treatment with Mastoparan-M (0.05, 0.1, and 0.2 mg/kg, IP) for 3 or 7 days, behavioral tests, H&E staining or Nissl staining, immunohistochemistry, and ELISA were employed to investigate neuroprotective effects of Mastoparan-M on GCI in rats. Results: In vitro, the growth of HT22 neuronal cells was restrained at concentrations of 30-300 µg/ml (at 24 hr, IC50=105.2 µg/ml; at 48 hr, IC50=46.81 µg/ml), and Mastoparan-M treatment (0.1,1 and 5 µg/ml) restrained apoptosis. In vivo, Mastoparan-M improved neurocognitive function and neuronal loss in the hippocampal CA1 area of rats. In addition, these effects were associated with the prevention of neuroinflammation, oxidative stress, and apoptosis.

Chemically synthesized mastoparan M, a tetradecapeptide toxin of venom (INLKAIAALAKKLL), was used in the experiments described. After addition of mastoparan M to cultures of mouse macrophages in vitro, tumour necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta) were detected in the culture fluids by 12 h and their highest accumulation was observed by 24 h. Mastoparan M induced increases in both TNF-alpha secretion and mRNA level at the same time. Nitrite levels, which reflect nitric oxide synthesis, were also found to increase in the macrophage cultures at 24 h after mastoparan M addition. In vivo studies showed that mastoparan M induced the formation and accumulation of TNF-alpha, IL-1beta and nitrite in the peritoneal exudates of mice much faster at 90 min, 120 min and 180 min after mastoparan M injection, respectively. Similarly, significant increases in myeloperoxidase activity, a marker for neutrophil and macrophage content, were observed in the peritoneal lavage cells after intraperitoneal injection of mastoparan M. However, induction of nitrite by mastoparan M was completely inhibited by simultaneous addition of antimouse TNF-alpha antibody to the macrophage cultures. These results suggest that modulation of both neutrophil and macrophage influx by mastoparan M may be conveyed through TNF-alpha and IL-1beta secretion accompanied by nitrite formation.